WO2024259374A2

WO2024259374A2 - Methods for providing intercellular communication

Info

Publication number: WO2024259374A2
Application number: PCT/US2024/034200
Authority: WO
Inventors: Tapash Jay SARKAR; Akira Yamamoto; Joseph PAZZI
Original assignee: Rethink64 Bionetworks Pbc
Current assignee: Rethink64 Bionetworks Pbc
Priority date: 2023-06-14
Filing date: 2024-06-14
Publication date: 2024-12-19
Anticipated expiration: 2025-12-14
Also published as: WO2024259374A3

Abstract

To introduce material to cells, contemporary medicinal constructs rely on the uptake mechanisms of the cell membrane. This puts major restrictions on the types of utilizable materials (e.g., charge compatible), specifications (e.g., 100 nanometer scale or less) and organizations (mostly simplistic spheroids); this is the regime of nanoparticles, protein/peptide conjugates etc. However, the focus and novelty of the innovation presented are constructs which can still achieve this membrane interaction to connect to cells yet the constructs themselves remain outside of the cell, thus establishing a network by which to transfer materials. These can surpass the aforementioned limitations as well as create entirely new application spaces as these new constructs enable different desired distribution patterns and exchanged material.

Description

METHODS FOR PROVIDING INTERCELLULAR COMMUNICATION

CROSS REFERENCE

[0001] This application claims the benefit of U.S. Application No. 63/508,229, filed June 14, 2023 and U.S. Application No. 63/519,814, filed August 15, 2023, which are hereby incorporated by reference in their entirety.

BACKGROUND

[0002] Network architecture is used for delivery in multiple areas of cargo distribution, from people, to products, to information. They often involve establishment of key router nodes or hubs that connect various recipients and allow for organized traffic and exchange of the cargo. This strategy has yet to be applied in terms of therapeutic delivery to cells and tissues, which primarily relies on scattering individual carriers - each of which has some probability of uptake into a single target cell.

INCORPORATION BY REFERENCE

[0003] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

BRIEF DESCRIPTION OF THE DRAWINGS

[0004] FIG. 1A and IB Various schematics of construct networks comprising of containment, infrastructure, networking and cells.

[0005] FIG. 2 Schematic of various construct networks. Top schematic shows a first embodiment of a router construct comprising of composite infrastructure having an internal infrastructure embedded within the containment layer (e.g., the infrastructure is embedded in the containment membrane and the infrastructure and the localizing element are fully surrounded by the containment membrane). The composite infrastructure further comprises a localizing element (e.g., a hydrogel) and the internal infrastructure (e.g., comprised of calcium phosphate). The composite infrastructure comprising the localizing element and a number of infrastructures (e.g., n=4) along with the enclosed containment membrane may be considered a large router construct. The middle schematic shows a second embodiment showing a containment membrane surrounding an internal infrastructure wherein the infrastructure is within the containment membrane. A number of the infrastructure contained in the containment membrane (e.g., n=4) is dispersed within the localizing element (e.g., hydrogel). In some instances, the second embodiment of the router construct optionally comprises a localizing element. The bottom schematic shows a third exemplary construct comprising a containment membrane surrounding an internal infrastructure wherein the infrastructure is within the containment membrane. The infrastructure contained in the containment membrane is dispersed within the dispersal element (e.g., hydrocolloid). In some instances, the second construct optionally comprises a dispersal element. “CaP” denoted calcium phosphate. [0006] FIG. 3 shows a depiction of multiple failure modes and obstacles faced by nanoparticle delivery and reliance on endocytotic uptake.

[0007] FIG. 4 Schematic of other router interactions. These router interactions include routers potentially secreting or emitting encapsulated drug particles (budding drug particles) as one of the delivery mechanisms for uptake by the cell.

[0008] FIG. 5 Solid body infrastructures with different types of lipid membrane containment on the order of 20 micron in diameter. Labeled by rhodamine dye and bodipy lipid dyes, a) Zwitterionic DOPC membranes containing water and sucrose bodies, b) Cationic DOTAP membranes containing calcium phosphate bodies, c) Anionic DOPG and containing calcium phosphate bodies, and d) comparable mixture 33% Zwitterionic DOPC:33% Cationic DOTAP: 33% Anionic DOPG membrane with a Calcium Phosphate solid body. Scale bar: 100 micron.

[0009] FIG. 6 Fluid body infrastructures with lipid membrane containment holding multiple types of desired biological molecule on the order of 1 to 100 micron in diameter. Depicted by the fluorescent surrogates Alexa Fluor 555 for small molecules and FITC-BSA for large molecules, a) and b) are 75% Zwitterionic DOPC:25% Anionic DOPG with a PBS fluid body, c) and d) are 75% Zwitterionic DOPC:25% Anionic DOPG with a DMEM fluid body, e) and f) are 100% Zwitterionic DOPC with a water fluid body. Scale bar: a-d) 50 micron, e-f) 20 micron.

[0010] FIG. 7 Semisolid body infrastructures with lipid membrane containment. 100% Zwitterionic DOPC with an agarose semisolid body on the order of 400 micron squared to 1 mm squared in area and 0.1 micron to 10 micron in thickness. Also utilized is octadecyl isocyanate anchoring, a) brightfield, b) FITC-Dextran large molecule contained within construct c) rhodamine membrane dye and d) overlay image. Scale bar: 50 micron.

[0011] FIG. 8 Combination of solid body infrastructures with different types of lipid membrane containment. Mixture of calcium phosphate solid bodies - small population with cationic DOTAP containment and a larger population with anionic DOPG containment. Also shown are brightfield and an overlay in left and right column respectively, a) - d) initial mixture of two different lipid contained calcium phosphate beads, e) - h) 12 hours later after bead have chance to mix and combine a) brightfield. Membrane and bead combination sharing showing by the dramatically increased prevalence of cationic DOTAP containment among the bead population after 12 hrs, indicating combination with the anionic DOPG containment beads. Arrows in h) show particularly poignant combinations where bead groups appear to be yellowish (indicating a combination of beads). Scale bar: 100 micron.

[0012] FIG. 9 Combination of liquid body infrastructures with different types of lipid membrane containment. Mixture of water+sucrose liquid bodies - one population with cationic DOTAP containment and a second population of anionic DOPG containment. Composite structures indicated by arrows. Scale bar: 25 micron.

[0013] FIG. 10 Combination of semisolid body infrastructures with different types of lipid membrane containment. Mixture of agarose semisolid bodies - one population of lager flakes (around 100 micron in size) with 90% Zwitterionic DOPC: 10% Anionic DOPG containment of FITC-Dextran large molecule surrogates and the other population of small flakes (around 10 micron in size) with 90% Zwitterionic DOPC: 10% Cationic DOTAP containment but no surrogate molecule, a) and b) shows initial mixture of the two while c) shows combined structure with surrogate molecule sharing after 6 minutes. Scale bar: a- c) 10 micron.

[0014] FIG. 11 Combination solid body infrastructure with semisolid body infrastructure all with lipid membrane containment. Composite structure of agarose flakes and calcium phosphate beads with DOPC lipid containment, a) shows brightfield image of single composite structure while b) shows a group, c) shows the green channel with the FITC Dextran surrogate large molecule maintained by the beads in the composite structure. Containment membrane shown by the lipid in d) reveals larger flake structure in the composite, e) shows the overlay image. Scale bar: a-e) 100 micron.

[0015] FIG. 12 Establishment of tubule networks with solid body structure and lipid membrane containment. 90% DOPC: 10% DOTAP membranes containing calcium phosphate bodies a) and b) show bright field of constructs effectively migrating and attaching to cell. Orange lipid staining in c) and d) shows emergence of tubule arrays connecting to cells and other constructs. Scale bar: a), c) 100 micron and b), d) 25 micron.

[0016] FIG. 13 Establishment of tubule networks with fluid body structure and lipid membrane containment. 75% DOPC:25% DOPG membranes containing water and sucrose fluid bodies a) shows brightfield and green lipid staining shows tubule formation in b) as depicted by arrows. Scale bar: 25 micron.

[0017] FIG. 14 Establishment of tubule networks with solid-semisolid combination body structure and lipid membrane containment. Composite structure of agarose flakes and calcium phosphate beads with DOPC lipid containment, a) shows brightfield and the lipid staining shows tubule formation in b) as depicted by arrows. Scale bar: 50 micron.

[0018] FIG. 15 Scanning electron microscope imaging of tubule network at different magnifications. “C” denotes constructs and arrows identify particularly long tubules. Scale bar: a-b) 100 micron c) 10 micron.

[0019] FIG. 16 Additional networking strategies, a) DOPC contained water and sucrose fluid structures with zinc oxide spiky nanoparticles with lipid tubule formation identified by arrows, b) DOPC contained zinc oxide spiky nanoparticles hardbody structures with lipid, c) DOPC contained water and sucrose fluid structures with TAT cell-penetrating peptides molecule forming lipid membrane openings for networking identified by arrows, d) Brightfield image fluorescence image 90% DOPC: 10%DOTAP contained calcium phosphate with PEG-3K local dehydrator e) shows transfer of GFP protein from cells producing it to bead networked to them - distal bead shown to still be empty as indicated by arrows. Scale bar: 50 micron.

[0020] FIG. 17 Transfer of molecules through tubule networks with solid body structure and lipid membrane containment, a) showing bright field b) showing transfer of orange lipid and c) showing transfer of FITC-Dextran large molecule surrogate. Scale bar: 50 micron. [0021] FIG. 18 Transfer of exogenous cargo molecules from constructs to cells across network.

Fluorescent test molecules of each class including a) and b) FITC-dextran large molecule, c) and d) Cy3- labeled miRNA, e) and f) Cy3-labeled anti-miRNA, g) and h) FAM-labeled siRNA, i) and j) GFP encoding mRNA, and k) and 1) a Cy3 -labeled miRNA combined with a GFP encoding mRNA to show dual delivery. Images show transfer from construct across the tubule array networks and to the cells for each test class. For the mRNA cases, fluorescence translated protein is observed in the cell and reverse transferred to the construct. Scale bars: 50 micron.

[0022] FIG. 19 Transfer of “endogenous” cargo molecules from cells to construct across network. Fluorescent test molecules for a) and b) GFP encoding mRNA and c) and d) CY3-labeled anti-miRNA are first delivered into the cell by alternative methods, and then shown to transfer to the construct. For mRNA case, fluorescence translated protein is observed in the cell. Scale bar: 50 micron.

[0023] FIG. 20A depicts a microscope image of a tubule network as described herein. FIG. 20B depicts a connection diagram of a tubule network as described herein that is parsable by graph theory algorithms.

[0024] FIG. 21A shows a histogram depicting frequency of connections per bead in an exemplary tubule network, as measured using a method described herein. FIG. 21B shows a histogram depicting frequency of connections per cell in a tubule network, as measured using a method described herein. [0025] FIG. 22 shows a histogram depicting frequency of lengths of connections (e.g., tubes) in an exemplary tubule network, as measured using a method described herein.

[0026] FIG. 23 shows a plot depicting bivariate statistics of the length vs. angle of projection of a connection (e.g., tube) in an exemplary tubule network.

[0027] FIG. 24 shows a plot depicting bivariate statistics of the average length vs. number of connections per bead in an exemplary tubule network.

[0028] FIG. 25 shows a histogram depicting frequency of angle values of connections in an exemplary tubule network, as measured using a method described herein.

[0029] FIG. 26 shows distributions of routers in the skin (dark to opaque spheroids in bright-field), router membrane lipids, and fluorescent mRNA cargo after subconjunctival injection. DAPI staining further identifies nuclei. Scale bar: 100 m.

[0030] FIG. 27 shows distributions of routers in the conjunctiva (dark to opaque spheroids in bright- field), router membrane lipids, and fluorescent mRNA cargo after subcutaneous injection. DAPI staining further identifies nuclei. Scale bar: 100 m.

[0031] FIG. 28 shows distributions of routers in the liver (dark to opaque spheroids in bright-field), router membrane lipids, and fluorescent mRNA cargo after direct liver injection. DAPI staining further identifies nuclei. Scale bar: 100 m.

[0032] FIG. 29 shows distributions of routers in the pancreas (dark to opaque spheroids in bright- field), router membrane lipids, and fluorescent mRNA cargo after direct pancreas injection. DAPI staining further identifies nuclei. Scale bar: 100 m. [0033] FIG. 30 shows network formation utilizing polystyrene infrastructure routers (glossy spheroidal objects in bright-field). The signal corresponds to labeled containment membrane lipid connecting and shared with cells. Scale bar: 100 m.

[0034] FIG. 31 shows network formation utilizing CaCO , infrastructure routers (clusters of opaque spheroidal objects in bright-field). The signal corresponds to labeled containment membrane lipid connecting and shared with cells. Scale bar: 100 pm.

[0035] FIG. 32 shows network formation utilizing SiC>2 infrastructure routers (clusters of opaque material in bright-field). The signal corresponds to labeled containment membrane lipid connecting and shared with cells. Scale bar: 100 pm.

[0036] FIG. 33 shows network formation utilizing calcium alginate flake infrastructure routers (largely translucent in bright-field). The signal corresponds to labeled containment membrane lipid connecting and shared with cells. Scale bar: 100 pm.

[0037] FIG. 34 shows network formation utilizing lithium iron phosphate infrastructure routers (opaque bodies in bright-field). The signal corresponds to labeled containment membrane lipid connecting and shared with cells. Scale bar: 500 pm.

[0038] FIG. 35 shows network formation utilizing SiCTC 18 infrastructure routers (opaque bodies in bright-field). The signal corresponds to labeled containment membrane lipid connecting and shared with cells. Scale bar: 100 pm.

[0039] FIG. 36 shows network formation utilizing 10 pm and below calcium phosphate infrastructure routers (opaque bodies in bright-field). The signal corresponds to labeled containment membrane lipid connecting and shared with cells. Scale bar: 100 pm.

[0040] FIG. 37 shows network formation utilizing calcium phosphate flake infrastructure routers (semi-translucent bodies in bright-field). The signal corresponds to labeled containment membrane lipid connecting and shared with cells. Scale bar: 500 pm.

[0041] FIG. 38 shows delivery to known low transfectability cell type MDA-MB-468 with routers (translucent spheroids in brightfield), router membrane lipids, and fluorescent mRNA cargo. A comparison to transfection of the same cargo volume with Lipofectamine™ 3000 is also shown. Scale bar: 50 pm.

[0042] FIG. 39 shows network formation with routers (bright round bodies in bright-field) on HCT116 cells. The middle panel shows containment membrane lipid connecting and shared with cells. The right panel signal shows distributed mRNA cargo. Scale bar 50 pm.

[0043] FIG. 40 shows network formation with routers (bright round bodies in bright-field) on MCFlOa cells. The signal corresponds to labeled containment membrane lipid connecting and shared with cells. Scale bar: 100 pm.

[0044] FIG. 41 shows network formation with routers (opaque round bodies in bright-field) on MCF7 cells. The signal corresponds to labeled containment membrane lipid connecting and shared with cells. Scale bar: 100 pm. [0045] FIG. 42 shows delivery with routers (opaque spheroids in bright-field) of a fluorescent TAMRA-peptide cargo at a concentration of 10 pM. Scale bar: Scale: 100 pm.

[0046] FIG. 43 shows fluorescence images showing the routers (circles) during early tubule formation (lines/protrusions) over 3 to 25 minutes. Scale: 500 pm.

[0047] FIG. 44 shows brightfield image showing the cells remain on the petri dish after detachment of the routers. Fluorescence image showing the mRNA cargo retained after transfer from the routers to the cells. Scale: 100 pm.

[0048] FIG. 45 shows overlay confocal images showing the encapsulation of the mRNA inside the membrane-coated routers. Scale bars: 50 pm.

[0049] FIG. 46 shows reconstructed confocal fluorescence z-stack showing XZ orthogonal projection. This reveals taut network tubule connections extending in 3D. Scale bar: 20 pm (Left), 20 pm (Right). The figure shows pearls/packets/particles of loaded cargo materials inside

[0050] FIG. 47 shows reconstructed confocal fluorescence z-stack images showing the XY maximum intensity projection. This also reveals pearls or packets of loaded cargo materials traveling down tubules. Scale bar: 20 pm (Left), 20 pm (Right).

[0051] FIG. 48 shows network formation with 70:30 DOPC/DOTAP (membrane: networking).

Routers (opaque round bodies in bright-field). The signal corresponds to labeled containment membrane lipid connecting and shared with cells. Scale bar: 100 pm.

[0052] FIG. 49 shows network formation with 80:20 DOPC/DOTAP (membrane: networking).

[0053] FIG. 50 shows network formation with 90: 10 DOPC/DOTAP (membrane: networking).

[0054] FIG. 51 shows network formation and delivery with routers (bright round bodies in bright- field) on HEK293 cells. The middle panel shows the containment membrane lipid connecting and shared with cells. The right panel shows distributed mRNA cargo. Scale bar 50 pm.

[0055] FIG. 52 shows particle size distribution of lipid-coated calcium phosphate routers in pm.

[0056] FIG. 53 shows free floating unencapsulated mRNA cargo after formulation with the router vs.

Lipofectamine. Measurements were taken on supernatant of the samples on a NanoDrop ND 100 Spectrophotometer, ThermoFisher.

DETAILED DESCRIPTION

[0057] In one aspect, the development of therapeutic routers which establish network topologies for organized traffic and exchange of therapeutic cargo are described herein.

[0058] As used herein, and unless otherwise specified, the term "about" or "approximately" means an acceptable error for a particular value as determined by one of ordinary skill in the art, which depends in part on how the value is measured or determined. In certain embodiments, the term "about" or " approximately" means within 1, 2, 3, or 4 standard deviations. In certain embodiments, the term "about" or "approximately" means within 20%, 15%, 10%, or 5% of a given value or range.

[0059] The term Zwitterionic lipid refers to any of a number of lipid species that exist in an overall neutral form at physiological pH.

[0060] In some embodiments, an intercellular router construct system described herein comprising, (i) a cell; (ii) an intercellular router construct which itself remains extracellular, comprising a containment layer, a networking element, and an internal infrastructure, and wherein the containment layer comprises lipids; and (iii) a network that connects the cell and the intercellular router construct. In some embodiments, the intercellular router construct optionally contains one or more cargo molecules. In some embodiments, the containment layer is an extracellular containment layer.

[0061] In some embodiments, an intercellular router construct system described herein comprising (i) an intercellular router construct, comprising a containment layer, an internal infrastructure, and a networking element, wherein the internal infrastructure is located within the containment layer (ii) a network connecting (a) a cell and the intercellular router construct, or (b) a plurality of intercellular router constructs; and (iii) optionally, a cargo molecule.

[0062] In some embodiments, an intercellular router construct system described herein comprising (i) a cargo molecule; (ii) an intercellular router construct comprising an internal infrastructure, a networking element, and a containment layer; (iii) a network to and/or between a cell and/or other intercellular router constructs; and (iv) an exchange of the cargo molecule.

[0063] In some embodiments, the internal infrastructure is located within the containment layer wherein the infrastructure is surrounded by the containment layer. In some embodiments, the internal infrastructure is located within the containment layer but is not fully surrounded by the containment layer. In some embodiments, the internal infrastructure is embedded in the containment layer but is not surrounded by the containment layer.

[0064] In some embodiments, provided herein is an intercellular router construct comprising: (a) a containment layer; (b) a localizing element; and (c)a composite infrastructure, wherein the composite infrastructure comprises at least a portion of the localizing element and an internal infrastructure; wherein the containment layer encloses the composite infrastructure.

[0065] In some embodiments the intracellular router construct system further comprises a localizing element or a dispersal element. In some embodiments, the localizing element serves to localize a set of router constructs. The localizing element may group the individual router constructs but does not directly connect to them. In some instances, the localizing element may group the individual router constructs and are directly connected to the individual routers. The localizing element may have access to the interior space of the infrastructure, thus, at least a portion of the localizing element and a portion of the infrastructure form a composite infrastructure. A localizing embodiment can group at least 2, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more individual routers. For example, tubules do not form to the localizing element but may form through the localizing element. In some embodiments, the dispersing element spreads out the router constructs but does not directly connect to them. In some embodiments, the localizing element or dispersing element does not have access to the interior space of the infrastructure. In some embodiments, the containment layer is between the infrastructure and the localizing element or the dispersal element. In some embodiments, the containment layer is not between the infrastructure and the localizing element or the dispersal element.

[0066] In some embodiments, the localizing or dispersal element and the internal infrastructure are in fluid communication. In some embodiments, the localizing or dispersal element and the internal infrastructure are not in fluid communication.

[0067] In some embodiments, when a networking element is formed between the intercellular router and a cell, the networking element comprises two ends and one end connects to the internal infrastructure and one end connects to the cell.

[0068] In some embodiments, the localizing element is a hydrogel (e.g., an agarose hydrogel, photocuring hydrogel, or a modified starch, or the like).

[0069] In some embodiments, the dispersal element is a hydrocolloid (common hydrocolloids include but are not limited to carboxy-methylcellulose, carrageenan, starch, pectin, or proteins with the ability to produce viscous liquid on hydration).

[0070] In some embodiments, a cargo molecule does not pass to a localizing element or to a dispersal element. In some embodiments, a localizing element does not pass to a cargo molecule. In some embodiments, a dispersal element does not pass to a cargo molecule.

[0071] In some embodiments, a containment layer described herein insulates the internal infrastructure from an extracellular environment and prevents the leakage, loss, or exposure of the cargo into the extracellular environment. In some embodiments, the containment layer comprises lipids. In some embodiments, the lipids are present in a lipid membrane on the external surface of the intercellular router construct. In some embodiments, the external surface of the intercellular router construct is covered by the membrane. In some embodiments, the lipids comprise a Zwitterionic head group, a cationic head group, or an anionic head group, or any combinations thereof. In some embodiments, the lipids comprise Zwitterionic lipids. In some embodiments, the lipids comprise Zwitterionic lipids and membrane dopant lipids, wherein the total lipid composition of the containment layer comprises at most 5 mol%, 10 mol%, 15 mol%, 20 mol%, or 25 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise Zwitterionic lipids and membrane dopant lipids, wherein the total lipid composition of the containment layer comprises at most 10 mol%, 15 mol%, or 20 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise Zwitterionic lipids and membrane dopant lipids, wherein the total lipid composition of the containment layer comprises at most 10 mol% or 20 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise Zwitterionic lipids and membrane dopant lipids, wherein the total lipid composition of the containment layer comprises at most 5 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise Zwitterionic lipids and membrane dopant lipids, wherein the total lipid composition of the containment layer comprises at most 10 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise Zwitterionic lipids and membrane dopant lipids, wherein the total lipid composition of the containment layer comprises at most 15 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise Zwitterionic lipids and membrane dopant lipids, wherein the total lipid composition of the containment layer comprises at most 20 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise Zwitterionic lipids and membrane dopant lipids, wherein the total lipid composition of the containment layer comprises at most 25 mol% of the membrane dopant lipids. In some embodiments, the containment layer is an extracellular containment layer.

[0072] In some embodiments, a lipid described herein comprises cationic lipids. In some embodiments, the lipids comprise cationic lipids and membrane dopant lipids, wherein the total lipid composition of the containment layer comprises at most 5 mol%, 10 mol%, 15 mol%, 20 mol%, or 25 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise cationic lipids and membrane dopant lipids, wherein the total lipid composition of the containment layer comprises at most 10 mol%, 15 mol%, or 20 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise cationic lipids and membrane dopant lipids, wherein the total lipid composition of the containment layer comprises at most 10 mol% or 20 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise cationic lipids and membrane dopant lipids, wherein the total lipid composition of the containment layer comprises at most 5 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise cationic lipids and membrane dopant lipids, wherein the total lipid composition of the containment layer comprises at most 10 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise cationic lipids and membrane dopant lipids, wherein the total lipid composition of the containment layer comprises at most 15 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise cationic lipids and membrane dopant lipids, wherein the total lipid composition of the containment layer comprises at most 20 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise cationic lipids and membrane dopant lipids, wherein the total lipid composition of the containment layer comprises at most 25 mol% of the membrane dopant lipids. [0073] In some embodiments, a containment layer described herein comprises a lipid membrane that comprises lipids. In some embodiments, a containment layer comprises lipids. In some embodiments, the lipids comprise at most 5 mol% of cationic lipids. In some embodiments, the lipids comprise at most 10 mol% of cationic lipids. In some embodiments, the lipids comprise at most 15 mol% of cationic lipids. In some embodiments, the lipids comprise at most 20 mol% of cationic lipids. In some embodiments, the lipids comprise at most 25 mol% of cationic lipids. In some embodiments, the lipids comprise at most 30 mol% of cationic lipids. In some embodiments, the lipids comprise at most 35 mol% of cationic lipids. In some embodiments, the lipids comprise at most 40 mol% of cationic lipids. In some embodiments, the lipids comprise at most 45 mol% of cationic lipids. In some embodiments, the lipids comprise at most 50 mol% of cationic lipids. In some embodiments, the lipids comprise at most 55 mol% of cationic lipids. In some embodiments, the lipids comprise at most 60 mol% of cationic lipids. In some embodiments, the lipids comprise at most 65 mol% of cationic lipids. In some embodiments, the lipids comprise at most 70 mol% of cationic lipids. In some embodiments, the lipids comprise at most 75 mol% of cationic lipids. In some embodiments, the lipids comprise at most 80 mol% of cationic lipids. In some embodiments, the lipids comprise at most 85 mol% of cationic lipids. In some embodiments, the lipids comprise at most 90 mol% of cationic lipids. In some embodiments, the lipids comprise at most 95 mol% of cationic lipids. In some embodiments, the lipids comprise at most 100 mol% of cationic lipids. In some embodiments, the lipids comprise at least 5 mol% of cationic lipids. In some embodiments, the lipids comprise at least 10 mol% of cationic lipids. In some embodiments, the lipids comprise at least 15 mol% of cationic lipids. In some embodiments, the lipids comprise at least 20 mol% of cationic lipids. In some embodiments, the lipids comprise at least 25 mol% of cationic lipids. In some embodiments, the lipids comprise at least 30 mol% of cationic lipids. In some embodiments, the lipids comprise at least 35 mol% of cationic lipids. In some embodiments, the lipids comprise at least 40 mol% of cationic lipids. In some embodiments, the lipids comprise at least 45 mol% of cationic lipids. In some embodiments, the lipids comprise at least 50 mol% of cationic lipids. In some embodiments, the lipids comprise at least 55 mol% of cationic lipids. In some embodiments, the lipids comprise at least 60 mol% of cationic lipids. In some embodiments, the lipids comprise at least 65 mol% of cationic lipids. In some embodiments, the lipids comprise at least 70 mol% of cationic lipids. In some embodiments, the lipids comprise at least 75 mol% of cationic lipids. In some embodiments, the lipids comprise at least 80 mol% of cationic lipids. In some embodiments, the lipids comprise at least 85 mol% of cationic lipids. In some embodiments, the lipids comprise at least 90 mol% of cationic lipids. In some embodiments, the lipids comprise at least 95 mol% of cationic lipids. In some embodiments, the lipids comprise about 5-95 mol% of cationic lipids. In some embodiments, the lipids comprise about 5-50 mol% of cationic lipids. In some embodiments, the lipids comprise about 25- 95 mol% of cationic lipids. In some embodiments, the lipids comprise about 25-75 mol% of cationic lipids. In some embodiments, the lipids comprise about 0-90 mol% of cationic lipids. In some embodiments, the lipids comprise about 50-95 mol% of cationic lipids. In some embodiments, the lipids comprise about 50-99 mol% of cationic lipids. In some embodiments, the lipids comprise about 75 mol% of cationic lipids. In some embodiments, the lipids comprise about 25 mol% of cationic lipids.

[0074] In some embodiments, a containment layer described herein comprises a lipid membrane that comprises lipids. In some embodiments, a containment layer comprises lipids. In some embodiments, the lipids comprise at most 5 mol% of anionic lipids. In some embodiments, the lipids comprise at most 10 mol% of anionic lipids. In some embodiments, the lipids comprise at most 15 mol% of anionic lipids. In some embodiments, the lipids comprise at most 20 mol% of anionic lipids. In some embodiments, the lipids comprise at most 25 mol% of anionic lipids. In some embodiments, the lipids comprise at most 30 mol% of anionic lipids. In some embodiments, the lipids comprise at most 35 mol% of anionic lipids. In some embodiments, the lipids comprise at most 40 mol% of anionic lipids. In some embodiments, the lipids comprise at most 45 mol% of anionic lipids. In some embodiments, the lipids comprise at most 50 mol% of anionic lipids. In some embodiments, the lipids comprise at most 55 mol% of anionic lipids. In some embodiments, the lipids comprise at most 60 mol% of anionic lipids. In some embodiments, the lipids comprise at most 65 mol% of anionic lipids. In some embodiments, the lipids comprise at most 70 mol% of anionic lipids. In some embodiments, the lipids comprise at most 75 mol% of anionic lipids. In some embodiments, the lipids comprise at most 80 mol% of anionic lipids. In some embodiments, the lipids comprise at most 85 mol% of anionic lipids. In some embodiments, the lipids comprise at most 90 mol% of anionic lipids. In some embodiments, the lipids comprise at most 95 mol% of anionic lipids. In some embodiments, the lipids comprise at most 100 mol% of anionic lipids. In some embodiments, the lipids comprise at least 5 mol% of anionic lipids. In some embodiments, the lipids comprise at least 10 mol% of anionic lipids. In some embodiments, the lipids comprise at least 15 mol% of anionic lipids. In some embodiments, the lipids comprise at least 20 mol% of anionic lipids. In some embodiments, the lipids comprise at least 25 mol% of anionic lipids. In some embodiments, the lipids comprise at least 30 mol% of anionic lipids. In some embodiments, the lipids comprise at least 35 mol% of anionic lipids. In some embodiments, the lipids comprise at least 40 mol% of anionic lipids. In some embodiments, the lipids comprise at least 45 mol% of anionic lipids. In some embodiments, the lipids comprise at least 50 mol% of anionic lipids. In some embodiments, the lipids comprise at least 55 mol% of anionic lipids. In some embodiments, the lipids comprise at least 60 mol% of anionic lipids. In some embodiments, the lipids comprise at least 65 mol% of anionic lipids. In some embodiments, the lipids comprise at least 70 mol% of anionic lipids. In some embodiments, the lipids comprise at least 75 mol% of anionic lipids. In some embodiments, the lipids comprise at least 80 mol% of anionic lipids. In some embodiments, the lipids comprise at least 85 mol% of anionic lipids. In some embodiments, the lipids comprise at least 90 mol% of anionic lipids. In some embodiments, the lipids comprise at least 95 mol% of anionic lipids. In some embodiments, the lipids comprise about 5-95 mol% of anionic lipids. In some embodiments, the lipids comprise about 5-50 mol% of anionic lipids. In some embodiments, the lipids comprise about 25- 95 mol% of anionic lipids. In some embodiments, the lipids comprise about 25-75 mol% of anionic lipids. In some embodiments, the lipids comprise about 0-90 mol% of anionic lipids. In some embodiments, the lipids comprise about 50-95 mol% of anionic lipids. In some embodiments, the lipids comprise about 50-99 mol% of anionic lipids. In some embodiments, the lipids comprise about 75 mol% of anionic lipids. In some embodiments, the lipids comprise about 25 mol% of anionic lipids.

[0075] In some embodiments, a containment layer described herein comprises a lipid membrane that comprises lipids. In some embodiments, a containment layer comprises lipids. In some embodiments, the lipids comprise at most 5 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 10 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 15 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 20 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 25 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 30 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 35 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 40 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 45 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 50 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 55 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 60 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 65 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 70 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 75 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 80 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 85 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 90 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 95 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at most 100 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 5 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 10 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 15 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 20 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 25 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 30 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 35 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 40 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 45 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 50 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 55 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 60 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 65 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 70 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 75 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 80 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 85 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 90 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise at least 95 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise about 5-95 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise about 5-50 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise about 25-95 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise about 25-75 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise about 0-90 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise about 50-95 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise about 50-99 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise about 75 mol% of Zwitterionic lipids. In some embodiments, the lipids comprise about 25 mol% of Zwitterionic lipids.

[0076] In some embodiments, a containment layer described herein comprises a lipid membrane that comprises lipids. In some embodiments, a containment layer comprises lipids. In some embodiments, the lipids comprise at most 5 mol% of dopant lipids. In some embodiments, the lipids comprise at most 10 mol% of dopant lipids. In some embodiments, the lipids comprise at most 15 mol% of dopant lipids. In some embodiments, the lipids comprise at most 20 mol% of dopant lipids. In some embodiments, the lipids comprise at most 25 mol% of dopant lipids. In some embodiments, the lipids comprise at most 30 mol% of dopant lipids. In some embodiments, the lipids comprise at most 35 mol% of dopant lipids. In some embodiments, the lipids comprise at most 40 mol% of dopant lipids. In some embodiments, the lipids composition described in Example 1, 2, 3, or 4. In some embodiments, described herein is a composition according to Examples 1, 2, 3, or 4. [0077] In some embodiments, a lipid described herein comprises an anionic lipid. In some embodiments, the lipids comprise anionic lipids and membrane dopant lipids, wherein the total composition of the containment layer comprises at most 5 mol%, 10 mol%, 15 mol%, 20 mol%, or 25 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise anionic lipids and membrane dopant lipids, wherein the total composition of the containment layer comprises at most 10 mol%, 15 mol%, or 20 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise anionic lipids and membrane dopant lipids, wherein the total composition of the containment layer comprises at most 10 mol% or 20 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise anionic lipids and membrane dopant lipids, wherein the total composition of the containment layer comprises at most 5 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise anionic lipids and membrane dopant lipids, wherein the total composition of the containment layer comprises at most 10 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise anionic lipids and membrane dopant lipids, wherein the total composition of the containment layer comprises at most 15 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise anionic lipids and membrane dopant lipids, wherein the total composition of the containment layer comprises at most 20 mol% of the membrane dopant lipids. In some embodiments, the lipids comprise anionic lipids and membrane dopant lipids, wherein the total composition of the containment layer comprises at most 25 mol% of the membrane dopant lipids.

[0078] In some embodiments, a lipid described herein comprises a Zwitterionic lipid, a cationic lipid, or an anionic lipid, or any combinations thereof, provided that the Zwitterionic lipid, the cationic lipid, or the anionic lipid are greater than or equal to 10 mol% of the total lipid content of the containment layer. In some embodiments, the lipids comprise Zwitterionic lipids, cationic lipids, or anionic lipids, or any combinations thereof, provided that the Zwitterionic lipids, cationic lipids, or anionic lipids are greater than or equal to 2.5 mol% of the total lipid content of the containment layer. In some embodiments, the lipids comprise Zwitterionic lipids, cationic lipids, or anionic lipids, or any combinations thereof, provided that the Zwitterionic lipids, cationic lipids, or anionic lipids are greater than or equal to 5 mol% of the total lipid content of the containment layer. In some embodiments, the lipids comprise Zwitterionic lipids, cationic lipids, or anionic lipids, or any combinations thereof, provided that the Zwitterionic lipids, cationic lipids, or anionic lipids are greater than or equal to 15 mol% of the total lipid content of the containment layer. In some embodiments, the lipids comprise Zwitterionic lipids, cationic lipids, or anionic lipids, or any combinations thereof, provided that the Zwitterionic lipids, cationic lipids, or anionic lipids are greater than or equal to 20 mol% of the total lipid content of the containment layer. In some embodiments, the lipids comprise Zwitterionic lipids, cationic lipids, or anionic lipids, or any combinations thereof, provided that the Zwitterionic lipids, cationic lipids, or anionic lipids are greater than or equal to 25 mol% of the total lipid content of the containment layer.

[0079] In some embodiments, the lipid membrane comprises a ratio of cationic to anionic lipids. In some embodiments, the ratio of cationic to anionic lipids is about 95:5, 90: 10, 85: 15, 80:20, 75:25, 70:30, 65:35, 60:40, 55:45, 50:50, 45:55, 40:60, 35:65, 30:70, 25:75, 20:80, 15:85, 10:90, or 5:95. In some embodiments, the lipid membrane comprises a 95:5 ratio of cationic to anionic lipids. In some embodiments, the lipid membrane comprises a 90: 10 ratio of cationic to anionic lipids. In some embodiments, the lipid membrane comprises a 85: 15 ratio of cationic to anionic lipids. In some embodiments, the lipid membrane comprises a 80:20 ratio of cationic to anionic lipids. In some embodiments, the lipid membrane comprises a 75:25 ratio of cationic to anionic lipids. In some embodiments, the lipid membrane comprises a 70:30 ratio of cationic to anionic lipids. In some embodiments, the lipid membrane comprises a 65:35 ratio of cationic to anionic lipids. In some embodiments, the lipid membrane comprises a 60:40 ratio of cationic to anionic lipids. In some embodiments, the lipid membrane comprises a 55:45 ratio of cationic to anionic lipids. In some embodiments, the lipid membrane comprises a 50:50 ratio of cationic to anionic lipids.

[0080] In some embodiments, a lipid described herein comprises a single positive charge, a double positive charge, a triple positive charge, or a polycationic charge. In some embodiments, the lipids comprise a double positive charge, a triple positive charge, or a poly cationic charge. In some embodiments, the lipids comprise a single positive charge or a double positive charge. In some embodiments, the lipids comprise a single positive charge. In some embodiments, the lipids comprise a double positive charge. In some embodiments, the lipids comprise a triple positive charge. In some embodiments, the lipids comprise a poly cationic charge.

[0081] In some embodiments, a lipid described herein comprises a single negative charge, a double negative charge, a triple negative charge, or a polyanionic charge. In some embodiments, the lipids comprise a double negative charge, a triple negative charge, or a polyanionic charge. In some embodiments, the lipids comprise a single negative charge, a double negative charge, or a polyanionic charge. In some embodiments, the lipids comprise a single negative charge or a double negative charge. In some embodiments, the lipids comprise a single negative charge. In some embodiments, the lipids comprise a double negative charge. In some embodiments, the lipids comprise a triple negative charge. In some embodiments, the lipids comprise a polyanionic charge.

[0082] In some embodiments, a lipid described herein comprises various head group sizes. In some embodiments, the head group sizes are about 40 to about 80 square Angstroms (A²). In some embodiments, the head group sizes are about 20 to about 80 square Angstroms (A²). In some embodiments, the head group sizes are about 40 to about 100 square Angstroms (A²). In some embodiments, the head group sizes are about 20 to about 100 square Angstroms (A²). In some embodiments, the head group sizes are about 20 to about 100 square Angstroms (A²). In some embodiments, the head group sizes are about 20 to about 120 square Angstroms (A²). In some embodiments, the head group sizes are about 20 square Angstroms (A²). In some embodiments, the head group sizes are about 40 square Angstroms (A²). In some embodiments, the head group sizes are about 60 square Angstroms (A²). In some embodiments, the head group sizes are about 80 square Angstroms (A²). In some embodiments, the head group sizes are about 100 square Angstroms (A²). In some embodiments, the head group sizes are about 120 square Angstroms (A²). In some embodiments, the head group sizes are at most 40 square Angstroms (A²). In some embodiments, the head group sizes are at most 60 square Angstroms (A²). In some embodiments, the head group sizes are at most 80 square Angstroms (A²). In some embodiments, the head group sizes are at most 100 square Angstroms (A²). In some embodiments, the head group sizes are at most 120 square Angstroms (A²).

[0083] In some embodiments, a lipid described herein comprises a shape. In some embodiments, the shape of the lipids comprise a cylindrical shape, a cone-shape, or an inverted cone-shape, or any combinations thereof. In some embodiments, the lipids vary in shape. In some embodiments, the shape of the lipids comprise a cylindrical shape, a cone-shape, or an inverted cone-shape. In some embodiments, the shape of the lipids comprise a cone-shape or an inverted cone-shape, or any combinations thereof. In some embodiments, the shape of the lipids comprise a cone-shape or an inverted cone-shape. In some embodiments, the shape of the lipids comprise a cylindrical shape or an inverted cone-shape, or any combinations thereof. In some embodiments, the shape of the lipids comprise a cylindrical shape or an inverted cone-shape. In some embodiments, the shape of the lipids comprise a cylindrical shape or a cone-shape, or any combinations thereof. In some embodiments, the shape of the lipids comprise a cylindrical shape or a cone-shape. In some embodiments, the shape of the lipids comprise a cylindrical shape. In some embodiments, the shape of the lipids comprise a cone-shape. In some embodiments, the shape of the lipids comprise an inverted cone-shape. In some embodiments, the shape of the lipids comprise an irregular shape. In some embodiments, the lipids vary in shape. In some embodiments, the shape of the lipids comprise a cylindrical shape, a cone-shape, an inverted cone-shape, or an irregular, or any combinations thereof. In some embodiments, the lipids vary in shape. In some embodiments, the shape of the lipids comprise a cylindrical shape, a cone-shape, an inverted cone-shape, or an irregular. [0084] In some embodiments, a lipid described herein comprises a phospholipid head group, a glycolipid head group, or a sterile head group, or any combinations thereof. In some embodiments, the lipids comprise a phospholipid head group, a glycolipid head group, or a sterile head group. In some embodiments, the lipids comprise a phospholipid head group or a glycolipid head group, or any combinations thereof. In some embodiments, the lipids comprise a phospholipid head group or a glycolipid head group. In some embodiments, the lipids comprise a phospholipid head group or a sterile head group, or any combinations thereof. In some embodiments, the lipids comprise a phospholipid head group or a sterile head group. In some embodiments, the lipids comprise a glycolipid head group or a sterile head group, or any combinations thereof. In some embodiments, the lipids comprise a glycolipid head group or a sterile head group. In some embodiments, the lipids comprise a phospholipid head group. In some embodiments, the lipids comprise a glycolipid head group. In some embodiments, the lipids comprise a sterile head group.

[0085] In some embodiments, a lipid described herein comprises various chain lengths, number of tails, or number of double bonds, or any combinations thereof. In some embodiments, the lipids comprise various chain lengths, number of tails, or number of double bonds. In some embodiments, the lipids comprise various number of tails or number of double bonds, or any combinations thereof. In some embodiments, the lipids comprise various number of tails or number of double bonds. In some embodiments, the lipids comprise various chain lengths or number of double bonds, or any combinations thereof. In some embodiments, the lipids comprise various chain lengths or number of double bonds. In some embodiments, the lipids comprise various chain lengths, number of tails, or number of double bonds, or any combinations thereof. In some embodiments, the lipids comprise various chain lengths, number of tails, or number of double bonds. In some embodiments, the lipids comprise various chain lengths. In some embodiments, the lipids comprise various number of tails. In some embodiments, the lipids comprise various number of double bonds.

[0086] In some embodiments, a lipid described herein comprises a polymerized lipid.

[0087] In some embodiments, a lipid described herein comprises a bilayer or a monolayer membrane, or any combination thereof. In some embodiments, the lipids comprise a bilayer or a monolayer membrane. In some embodiments, the lipids comprise a bilayer membrane. In some embodiments, the lipids comprise a monolayer membrane. In some embodiments, the lipids are unilamellar or multilamellar. In some embodiments, the lipids are unilamellar. In some embodiments, the lipids are multilamellar.

[0088] In some embodiments, a lipid described herein comprises a Zwitterionic lipid, a cationic lipid, or an anionic lipid, or any combinations thereof. In some embodiments, a lipid described herein comprises soy phosphatidylcholine (Soy PC), dioleoyl phosphatidylcholine (DOPC), dioleoyl phosphatidylglycerol (DOPG), dioleoyl phosphatidylethanolamine (DOPE), dioleoyl trimethylammonium -propane (DOTAP), dioleyloxy-N-sperminecarboxamido ethyl -N, N-dimethyl- 1- propanaminium (DOSPA), dipalmitoylphosphatidylcholine (DPPC), hydrogenated soy phosphatidylcholine (HSPC), or heptatriacont tetraene dimethylamino butanoate (DLin-DMA), or any combination thereof. In some embodiments, the lipids comprise soy phosphatidylcholine (Soy PC), dioleoyl phosphatidylcholine (DOPC), dioleoyl phosphatidylglycerol (DOPG), dioleoyl phosphatidylethanolamine (DOPE), dioleoyl trimethylammonium-propane (DOTAP), or dioleyloxy-N- sperminecarboxamido ethyl-N, N-dimethyl- 1 -propanaminium (DOSPA), or any combination thereof. In some embodiments, the lipids comprise dioleoyl phosphatidylcholine (DOPC), dioleoyl phosphatidylglycerol (DOPG), dioleoyl phosphatidylethanolamine (DOPE), or dioleoyl trimethylammonium-propane (DOTAP), or any combination thereof. In some embodiments, the lipids comprise dioleoyl phosphatidylcholine (DOPC), dioleoyl phosphatidylglycerol (DOPG), or dioleoyl trimethylammonium-propane (DOTAP), or any combination thereof. In some embodiments, the lipids comprise dioleoyl phosphatidylcholine (DOPC), dioleoyl phosphatidylglycerol (DOPG), or dioleoyl trimethylammonium-propane (DOTAP). In some embodiments, the lipids comprise dioleoyl phosphatidylcholine (DOPC), or dioleoyl trimethylammonium-propane (DOTAP), or a combination thereof. In some embodiments, the lipids comprise a combination of dioleoyl phosphatidylcholine (DOPC) and dioleoyl trimethylammonium-propane (DOTAP).

[0089] In some embodiments, a containment layer described herein comprises a polymer, a metallic substrate, a plastic, a ceramics, or a fatty acid, or any combinations thereof. In some embodiments, the containment layer comprises a polymer, a metallic substrate, a plastic, a ceramics, or a fatty acid. In some embodiments, the containment layer comprises a metallic substrate, a plastic, or a ceramics, or any combinations thereof. In some embodiments, the containment layer comprises a metallic substrate, a plastic, or a ceramics. In some embodiments, the containment layer comprises a polymer, a metallic substrate, or a fatty acid, or any combinations thereof. In some embodiments, the containment layer comprises a polymer, a metallic substrate, or a fatty acid. In some embodiments, the containment layer comprises a polymer, a metallic substrate, or a plastic, or any combinations thereof. In some embodiments, the containment layer comprises a polymer, a metallic substrate, or a plastic. In some embodiments, the containment layer comprises a polymer. In some embodiments, the containment layer comprises a metallic substrate. In some embodiments, the containment layer comprises a plastic. In some embodiments, the containment layer comprises a ceramics. In some embodiments, the containment layer comprises a fatty acid.

[0090] In some embodiments, a containment layer described herein comprises a plurality of independent intercellular router constructs. In some embodiments, the containment layers of the plurality of intercellular router constructs are independently selected from a lipid, a polymer, a metallic substrate, a plastic, a ceramic, and a fatty acid. In some embodiments, the containment layers of the plurality of intercellular router constructs are independently selected from a lipid, a polymer, a metallic substrate, a plastic, and a ceramic. In some embodiments, the containment layers of the plurality of intercellular router constructs are independently selected from a lipid, a polymer, a metallic substrate, and a ceramic. In some embodiments, the containment layers of the plurality of intercellular router constructs are independently selected from a lipid, a polymer, and a metallic substrate. In some embodiments, the containment layers of the plurality of intercellular router constructs are independently selected from a lipid and a metallic substrate. In some embodiments, the containment layers of the plurality of intercellular router constructs are independently selected from a lipid. In some embodiments, the containment layers of the plurality of intercellular router constructs are independently selected from a polymer. In some embodiments, the containment layers of the plurality of intercellular router constructs are independently selected from aa metallic substrate. In some embodiments, the containment layers of the plurality of intercellular router constructs are independently selected from a plastic. In some embodiments, the containment layers of the plurality of intercellular router constructs are independently selected from a ceramic. In some embodiments, the containment layers of the plurality of intercellular router constructs are independently selected from a fatty acid. In some embodiments, the plurality of intercellular router constructs comprise a cationic lipid containment layer and an anionic lipid containment layer. In some embodiments, the plurality of intercellular router constructs comprise a cationic lipid containment layer. In some embodiments, the plurality of intercellular router constructs comprise an anionic lipid containment layer.

[0091] In some embodiments, an internal infrastructure described herein comprises a geometry, a scale, a phase, a porosity, or a surface charge, or any combinations thereof. In some embodiments, the internal infrastructure comprises a geometry, a scale, a phase, a porosity, or a surface charge. In some embodiments, the internal infrastructure comprises a geometry, a scale, a phase, or a surface charge, or any combinations thereof. In some embodiments, the internal infrastructure comprises a geometry, a scale, a phase, or a surface charge. In some embodiments, the internal infrastructure comprises a geometry, a scale, a phase, or a porosity, or any combinations thereof. In some embodiments, the internal infrastructure comprises a geometry, a scale, a phase, or a porosity. In some embodiments, the internal infrastructure comprises a geometry, a scale, or a phase, or any combinations thereof. In some embodiments, the internal infrastructure comprises a geometry, a scale, or a phase. In some embodiments, the internal infrastructure comprises a geometry. In some embodiments, the internal infrastructure comprises a scale. In some embodiments, the internal infrastructure comprises a phase. In some embodiments, the internal infrastructure comprises a porosity. In some embodiments, the internal infrastructure comprises a surface charge. In some embodiments, the internal infrastructure comprises a surface charge potential. In some embodiments, the surface charge potential is positive. In some embodiments, the surface charge potential is negative. In some embodiments, the surface charge is neutral.

[0092] In some embodiments, an internal infrastructure described herein comprises a spheroidal geometry, a planar geometry, a tubular geometry, or an irregular geometry. In some embodiments, the internal infrastructure comprises a spheroidal geometry, a planar geometry, or a tubular geometry. In some embodiments, the internal infrastructure comprises a spheroidal geometry, a tubular geometry, or an irregular geometry. In some embodiments, the internal infrastructure comprises a planar geometry, a tubular geometry, or an irregular geometry. In some embodiments, the internal infrastructure comprises a spheroidal geometry. In some embodiments, the internal infrastructure comprises a planar geometry. In some embodiments, the internal infrastructure comprises a tubular geometry. In some embodiments, the internal infrastructure comprises an irregular geometry.

[0093] In some embodiments, an internal infrastructure described herein is about 10 nm to 10 mm. In some embodiments, the internal infrastructure is about 50 nm to 10 mm. In some embodiments, the internal infrastructure is about 100 nm to 10 mm. In some embodiments, the internal infrastructure is about 150 nm to 10 mm. In some embodiments, the internal infrastructure is about 200 nm to 10 mm. In some embodiments, the internal infrastructure is about 250 nm to 10 mm. In some embodiments, the internal infrastructure is about 300 nm to 10 mm. In some embodiments, the internal infrastructure is about 350 nm to 10 mm. In some embodiments, the internal infrastructure is about 400 nm to 10 mm. In some embodiments, the internal infrastructure is about 450 nm to 10 mm. In some embodiments, internal infrastructure is about 500 nm to 10 mm. In some embodiments, internal infrastructure is about 500 nm to 10 mm. In some embodiments, internal infrastructure is about 600 nm to 10 mm. In some embodiments, internal infrastructure is about 700 nm to 10 mm. In some embodiments, internal infrastructure is about 800 nm to 10 mm. In some embodiments, internal infrastructure is about 900 nm to 10 mm. In some embodiments, internal infrastructure is about 1000 nm to 10 mm. In some embodiments, internal infrastructure is about 10 pm to 10 mm. In some embodiments, internal infrastructure is about 50 pm to 10 mm. In some embodiments, internal infrastructure is about 100 pm to 10 mm. In some embodiments, internal infrastructure is about 200 pm to 10 mm. In some embodiments, internal infrastructure is about 300 pm to 10 mm. In some embodiments, internal infrastructure is about 400 gm to 10 mm. In some embodiments, internal infrastructure is about 500 gm to 10 mm. In some embodiments, the internal infrastructure is at most 10 nm. In some embodiments, the internal infrastructure is at most 50 nm. In some embodiments, the internal infrastructure is at most 100 nm. In some embodiments, the internal infrastructure is at most 150 nm. In some embodiments, the internal infrastructure is at most 200 nm. In some embodiments, the internal infrastructure is at most 250 nm. In some embodiments, the internal infrastructure is at most 300 nm. In some embodiments, the internal infrastructure is at most 450 nm. In some embodiments, the internal infrastructure is at most 500 nm. In some embodiments, the internal infrastructure is at most 600 nm. In some embodiments, the internal infrastructure is at most 700 nm. In some embodiments, the internal infrastructure is at most 800 nm. In some embodiments, the internal infrastructure is at most 900 nm. In some embodiments, the internal infrastructure is at most 1000 nm. In some embodiments, the internal infrastructure is at most 15 mm. In some embodiments, the internal infrastructure is at most 10 mm. In some embodiments, the internal infrastructure is at most 5 mm. In some embodiments, the internal infrastructure is at most 1 mm. In some embodiments, the internal infrastructure is about 10 nm to 1 mm. In some embodiments, the internal infrastructure is about 10 nm to 5 mm. In some embodiments, the internal infrastructure is about 10 nm to 15 mm.

[0094] In some embodiments, an internal infrastructure described herein is about 10 nm to 10 mm in largest dimension. In some embodiments, the internal infrastructure is about 50 nm to 10 mm in largest dimension. In some embodiments, the internal infrastructure is about 100 nm to 10 mm in largest dimension. In some embodiments, the internal infrastructure is about 150 nm to 10 mm in largest dimension. In some embodiments, the internal infrastructure is about 200 nm to 10 mm in largest dimension. In some embodiments, the internal infrastructure is about 250 nm to 10 mm in largest dimension. In some embodiments, the internal infrastructure is about 300 nm to 10 mm in largest dimension. In some embodiments, the internal infrastructure is about 350 nm to 10 mm in largest dimension. In some embodiments, the internal infrastructure is about 400 nm to 10 mm in largest dimension. In some embodiments, the internal infrastructure is about 450 nm to 10 mm in largest dimension. In some embodiments, internal infrastructure is about 500 nm to 10 mm in largest dimension. In some embodiments, internal infrastructure is about 500 nm to 10 mm in largest dimension. In some embodiments, internal infrastructure is about 600 nm to 10 mm in largest dimension. In some embodiments, internal infrastructure is about 700 nm to 10 mm in largest dimension. In some embodiments, internal infrastructure is about 800 nm to 10 mm in largest dimension. In some embodiments, internal infrastructure is about 900 nm to 10 mm in largest dimension. In some embodiments, internal infrastructure is about 1000 nm to 10 mm in largest dimension. In some embodiments, internal infrastructure is about 10 gm to 10 mm in largest dimension. In some embodiments, internal infrastructure is about 50 gm to 10 mm in largest dimension. In some embodiments, internal infrastructure is about 100 gm to 10 mm in largest dimension. In some embodiments, internal infrastructure is about 200 gm to 10 mm in largest dimension. In some embodiments, internal infrastructure is about 300 gm to 10 mm in largest dimension. In some embodiments, internal infrastructure is about 400 gm to 10 mm in largest dimension. In some embodiments, internal infrastructure is about 500 gm to 10 mm in largest dimension. In some embodiments, the internal infrastructure is at most 10 nm in largest dimension. In some embodiments, the internal infrastructure is at most 50 nm in largest dimension. In some embodiments, the internal infrastructure is at most 100 nm in largest dimension. In some embodiments, the internal infrastructure is at most 150 nm in largest dimension. In some embodiments, the internal infrastructure is at most 200 nm in largest dimension. In some embodiments, the internal infrastructure is at most 250 nm in largest dimension. In some embodiments, the internal infrastructure is at most 300 nm in largest dimension. In some embodiments, the internal infrastructure is at most 450 nm in largest dimension. In some embodiments, the internal infrastructure is at most 500 nm in largest dimension. In some embodiments, the internal infrastructure is at most 600 nm in largest dimension. In some embodiments, the internal infrastructure is at most 700 nm in largest dimension. In some embodiments, the internal infrastructure is at most 800 nm in largest dimension. In some embodiments, the internal infrastructure is at most 900 nm in largest dimension. In some embodiments, the internal infrastructure is at most 1000 nm in largest dimension. In some embodiments, the internal infrastructure is at most 15 mm in largest dimension. In some embodiments, the internal infrastructure is at most 10 mm in largest dimension. In some embodiments, the internal infrastructure is at most 5 mm in largest dimension. In some embodiments, the internal infrastructure is at most 1 mm in largest dimension. In some embodiments, the internal infrastructure is about 10 nm to 1 mm in largest dimension. In some embodiments, the internal infrastructure is about 10 nm to 5 mm in largest dimension. In some embodiments, the internal infrastructure is about 10 nm to 15 mm in largest dimension.

[0095] In some embodiments, an internal infrastructure described herein is about 10 nm to 10 mm in diameter. In some embodiments, the internal infrastructure is about 50 nm to 10 mm in diameter. In some embodiments, the internal infrastructure is about 100 nm to 10 mm in diameter. In some embodiments, the internal infrastructure is about 150 nm to 10 mm in diameter. In some embodiments, the internal infrastructure is about 200 nm to 10 mm in diameter. In some embodiments, the internal infrastructure is about 250 nm to 10 mm in diameter. In some embodiments, the internal infrastructure is about 300 nm to 10 mm in diameter. In some embodiments, the internal infrastructure is about 350 nm to 10 mm in diameter. In some embodiments, the internal infrastructure is about 400 nm to 10 mm in diameter. In some embodiments, the internal infrastructure is about 450 nm to 10 mm in diameter. In some embodiments, internal infrastructure is about 500 nm to 10 mm in diameter. In some embodiments, internal infrastructure is about 600 nm to 10 mm in diameter. In some embodiments, internal infrastructure is about 700 nm to 10 mm in diameter. In some embodiments, internal infrastructure is about 800 nm to 10 mm in diameter. In some embodiments, internal infrastructure is about 900 nm to 10 mm in diameter. In some embodiments, internal infrastructure is about 1000 nm to 10 mm in diameter. In some embodiments, internal infrastructure is about 500 nm to 1 gm mm in diameter. In some embodiments, internal infrastructure is about 500 nm to 5 gm mm in diameter. In some embodiments, internal infrastructure is about 10 gm to 10 mm in diameter. In some embodiments, internal infrastructure is about 50 |im to 10 mm in diameter. In some embodiments, internal infrastructure is about 100 gm to 10 mm in diameter. In some embodiments, internal infrastructure is about 200 |im to 10 mm in diameter. In some embodiments, internal infrastructure is about 300 gm to 10 mm in diameter. In some embodiments, internal infrastructure is about 400 gm to 10 mm in diameter. In some embodiments, internal infrastructure is about 500 |im to 10 mm in diameter. In some embodiments, the internal infrastructure is at most 10 nm in diameter. In some embodiments, the internal infrastructure is at most 50 nm in diameter. In some embodiments, the internal infrastructure is at most 100 nm in diameter. In some embodiments, the internal infrastructure is at most 150 nm in diameter. In some embodiments, the internal infrastructure is at most 200 nm in diameter. In some embodiments, the internal infrastructure is at most 250 nm in diameter. In some embodiments, the internal infrastructure is at most 300 nm in diameter. In some embodiments, the internal infrastructure is at most 450 nm in diameter. In some embodiments, the internal infrastructure is at most 500 nm in diameter. In some embodiments, the internal infrastructure is at most 600 nm in diameter. In some embodiments, the internal infrastructure is at most 700 nm in diameter. In some embodiments, the internal infrastructure is at most 800 nm in diameter. In some embodiments, the internal infrastructure is at most 900 nm in diameter. In some embodiments, the internal infrastructure is at most 1000 nm in diameter. In some embodiments, the internal infrastructure is at most 15 mm in diameter. In some embodiments, the internal infrastructure is at most 10 mm in diameter. In some embodiments, the internal infrastructure is at most 5 mm in diameter. In some embodiments, the internal infrastructure is at most 1 mm in diameter. In some embodiments, the internal infrastructure is about 10 nm to 1 mm in diameter. In some embodiments, the internal infrastructure is about 10 nm to 5 mm in diameter. In some embodiments, the internal infrastructure is about 10 nm to 15 mm in diameter.

[0096] In some embodiments, an internal infrastructure described herein is about 10 nm to 10 mm in scale. In some embodiments, the internal infrastructure is about 50 nm to 10 mm in scale. In some embodiments, the internal infrastructure is about 100 nm to 10 mm in scale. In some embodiments, the internal infrastructure is about 150 nm to 10 mm in scale. In some embodiments, the internal infrastructure is about 200 nm to 10 mm in scale. In some embodiments, the internal infrastructure is about 250 nm to 10 mm in scale. In some embodiments, the internal infrastructure is about 300 nm to 10 mm in scale. In some embodiments, the internal infrastructure is about 350 nm to 10 mm in scale. In some embodiments, the internal infrastructure is about 400 nm to 10 mm in scale. In some embodiments, the internal infrastructure is about 450 nm to 10 mm in scale. In some embodiments, internal infrastructure is about 500 nm to 10 mm in scale. In some embodiments, internal infrastructure is about 600 nm to 10 mm in scale. In some embodiments, internal infrastructure is about 700 nm to 10 mm in scale. In some embodiments, internal infrastructure is about 800 nm to 10 mm in scale. In some embodiments, internal infrastructure is about 900 nm to 10 mm in scale. In some embodiments, internal infrastructure is about 1000 nm to 10 mm in scale. In some embodiments, internal infrastructure is about 10 n to 10 mm in scale. In some embodiments, internal infrastructure is about 50 |im to 10 mm in scale. In some embodiments, internal infrastructure is about 100 gm to 10 mm in scale. In some embodiments, intemal infrastructure is about 200 pm to 10 mm in scale. In some embodiments, internal infrastructure is about 300 pm to 10 mm in scale. In some embodiments, internal infrastructure is about 400 gm to 10 mm in scale. In some embodiments, internal infrastructure is about 500 pm to 10 mm in scale. In some embodiments, the internal infrastructure is at most 10 nm in scale. In some embodiments, the internal infrastructure is at most 50 nm in scale. In some embodiments, the internal infrastructure is at most 100 nm in scale. In some embodiments, the internal infrastructure is at most 150 nm in scale. In some embodiments, the internal infrastructure is at most 200 nm in scale. In some embodiments, the internal infrastructure is at most 250 nm in scale. In some embodiments, the internal infrastructure is at most 300 nm in scale. In some embodiments, the internal infrastructure is at most 450 nm in scale. In some embodiments, the internal infrastructure is at most 500 nm in scale. In some embodiments, the internal infrastructure is at most 600 nm in scale. In some embodiments, the internal infrastructure is at most 700 nm in scale. In some embodiments, the internal infrastructure is at most 800 nm. In some embodiments, the internal infrastructure is at most 900 nm in scale. In some embodiments, the internal infrastructure is at most 1000 nm in scale. In some embodiments, the internal infrastructure is at most 15 mm in scale. In some embodiments, the internal infrastructure is at most 10 mm in scale. In some embodiments, the internal infrastructure is at most 5 mm in scale. In some embodiments, the internal infrastructure is at most 1 mm in scale. In some embodiments, the internal infrastructure is about 10 nm to 1 mm in scale. In some embodiments, the internal infrastructure is about 10 nm to 5 mm in scale. In some embodiments, the internal infrastructure is about 10 nm to 15 mm in scale.

[0097] In some embodiments, the internal infrastructure may be of varying size and shape. In some embodiments, the internal infrastructure has a particle size distribution. In some embodiments, the particle size distribution is from about 10 pm to about 35 pm. In some embodiments, the particle size distribution is from about 15 pm to about 30 pm. In some embodiments, the particle size distribution is from about 19 pm to about 27 pm. In some embodiments, the particle size distribution is from about 21 pm to about 25 pm.

[0098] In some embodiments, an internal infrastructure described herein comprises a solid body phase, a fluid body phase, or a semi-sold body phase. In some embodiments, the internal infrastructure comprises a solid body phase or a semi-sold body phase. In some embodiments, the internal infrastructure comprises fluid body phase or a semi-sold body phase. In some embodiments, the internal infrastructure comprises a solid body phase or a fluid body phase. In some embodiments, the internal infrastructure comprises a solid body phase. In some embodiments, the internal infrastructure comprises a fluid body phase. In some embodiments, the internal infrastructure comprises a semi-sold body phase. In some embodiments, the solid body phase comprises an insoluble solid. In some embodiments, the insoluble solid comprises a solid polymer, a salt, a metal, or a metal oxide. In some embodiments, the insoluble solid comprises a salt, a metal, or a metal oxide. In some embodiments, the insoluble solid comprises a solid polymer or a salt. In some embodiments, the insoluble solid comprises a solid polymer. In some embodiments, the insoluble solid is a salt. [0099] In some embodiments, a salt described herein is a metal salt. In some embodiments, the metal salt comprises a divalent cation. In some embodiments, the metal salt comprises a calcium divalent cation or a magnesium divalent cation. In some embodiments, the metal salt comprises a calcium divalent cation. In some embodiments, the metal salt comprises a magnesium divalent cation. In some embodiments, the metal salt comprises a phosphate anion, a carbonate anion, or a sulfate anion, or any combinations thereof. In some embodiments, the metal salt comprises a phosphate anion, a carbonate anion, or a sulfate anion. In some embodiments, the metal salt comprises a phosphate anion or a carbonate anion, or any combinations thereof. In some embodiments, the metal salt comprises a phosphate anion or a carbonate anion. In some embodiments, the metal salt comprises a phosphate anion or a sulfate anion, or any combinations thereof. In some embodiments, the metal salt comprises a phosphate anion or a sulfate anion. In some embodiments, the metal salt comprises a carbonate anion or a sulfate anion, or any combinations thereof. In some embodiments, the metal salt comprises a carbonate anion or a sulfate anion. In some embodiments, the metal salt comprises a phosphate anion. In some embodiments, the metal salt comprises a carbonate anion. In some embodiments, the metal salt comprises a sulfate anion. In some embodiments, the metal salt is calcium phosphate. In some embodiments, the calcium phosphate is a calcium phosphate flake. In some embodiments, the metal salt is a calcium carbonate. In some embodiments, the metal salt is magnesium phosphate.

[00100] In some embodiments, the metal salt is calcium alginate.

[00101] In some embodiments, the metal salt is lithium iron phosphate.

[00102] In some embodiments, an insoluble solid described herein is a metal. In some embodiments, the insoluble solid is a metal oxide. In some embodiments, the metal oxide is silica oxide or zinc oxide, or any combinations thereof. In some embodiments, the metal oxide is silica oxide or zinc oxide. In some embodiments, the metal oxide is silica oxide. In some embodiments, the metal oxide is silica dioxide. In some embodiments, the metal oxide is zinc oxide.

[00103] In some embodiments, a solid body phase described herein comprises an insoluble solid, a metal, a metal oxide, or a solid polymer, or any combinations thereof. In some embodiments, the solid body phase comprises an insoluble solid, a metal, a metal oxide, or a solid polymer. In some embodiments, the solid body phase comprises an insoluble solid, a metal, or a metal oxide, or any combinations thereof. In some embodiments, the solid body phase comprises an insoluble solid, a metal, or a metal oxide. In some embodiments, the solid body phase comprises an insoluble solid or a solid polymer, or any combinations thereof. In some embodiments, the solid body phase comprises an insoluble solid or a solid polymer. In some embodiments, the solid body phase comprises an insoluble solid. In some embodiments, the solid body phase comprises a metal. In some embodiments, the solid body phase comprises a metal oxide. In some embodiments, the solid body phase comprises a solid polymer.

[00104] In some embodiments, an infrastructure described herein comprises a fluid body phase. In some embodiments, the fluid body phase comprises a physiological buffer. In some embodiments, the physiological buffer comprises a salt, a nutrient, a sugar, an amino acid, or a vitamin, or any combinations thereof. In some embodiments, the physiological buffer comprises a salt, a nutrient, a sugar, an amino acid, or a vitamin. In some embodiments, the physiological buffer comprises a salt, a nutrient, a sugar, or an amino acid, or any combinations thereof. In some embodiments, the physiological buffer comprises a salt, a nutrient, or a sugar, or any combinations thereof. In some embodiments, the physiological buffer comprises a salt or a sugar, or any combinations thereof. In some embodiments, the physiological buffer comprises a salt, a sugar, an amino acid, or a vitamin, or any combinations thereof. In some embodiments, the physiological buffer comprises a salt, a sugar, or an amino acid, or any combinations thereof. In some embodiments, the physiological buffer comprises a salt. In some embodiments, the physiological buffer comprises a nutrient. In some embodiments, the physiological buffer comp rises a sugar. In some embodiments, the physiological buffer comprises an amino acid. In some embodiments, the physiological buffer comprises a vitamin. In some embodiments, the salt is LiCl, NaCl, KC1, or CsCl. In some embodiments, the salt is LiCl, NaCl, or KC1. In some embodiments, the salt is LiCl or NaCl. In some embodiments, the salt is NaCl. In some embodiments, the salt is LiCl.

[00105] In some embodiments, a physiological buffer described herein comprises a pH regulator. In some embodiments, the pH regulator comprises a phosphate salt, a carbonate salt, or a sulfate salt, or any combinations thereof. In some embodiments, the pH regulator comprises a phosphate salt or a carbonate salt, or any combinations thereof. In some embodiments, the pH regulator comprises a phosphate salt. In some embodiments, the pH regulator comprises a carbonate salt. In some embodiments, the pH regulator comprises a sulfate salt. In some embodiments, the physiological buffer comprises a minimal essential media or an equivalent cell supporting basal media. In some embodiments, the physiological buffer comprises a minimal essential media. In some embodiments, the physiological buffer comprises an equivalent cell supporting basal media.

[00106] In some embodiments, a fluid body phase described herein comprises a physiological buffer or a nontoxic buffer, or any combinations thereof. In some embodiments, the fluid body phase comprises a physiological buffer or a nontoxic buffer. In some embodiments, the fluid body phase comprises a nontoxic buffer. In some embodiments, the fluid body phase comprises a physiological buffer.

[00107] In some embodiments, an internal infrastructure described herein comprises a semi-solid body phase. In some embodiments, the semi-solid body phase comprises a cross-linkage. In some embodiments, the cross-linkage comprises a physical cross-linkage or a chemical cross-linkage, or any combinations thereof. In some embodiments, the cross-linkage comprises a physical cross-linkage or a chemical cross-linkage. In some embodiments, the cross-linkage comprises a physical cross-linkage. In some embodiments, the cross-linkage comprises a chemical cross-linkage.

[00108] In some embodiments, a semi-solid body phase described herein comprises a hydrogel. In some embodiments, the hydrogel comprises agarose, hyaluronans, chitosans, collagen, dextran, pectin, polylysine, gelatin, starch, polyvinylalcohol, poly(lactic-co-glycolic)acid (PLGA) polymers, (meth)acrylate-oligolactide-PEO-oligolactide-(meth)acrylate, polyethylene glycol) (PEO), polypropylene glycol) (PPO), PEO-PPO-PEO copolymers (Pluronic®), poly(phosphazene), poly(methacrylates), poly(N-vinylpyrrolidone), PL(G)A-PEO-PL(G)A copolymers, or poly(ethylene imine), or any combinations thereof. In some embodiments, the hydrogel comprises agarose, hyaluronans, chitosans, collagen, dextran, pectin, polylysine, gelatin, starch, polyvinylalcohol, poly(lactic-co-glycolic)acid (PLGA) polymers, (meth)acrylate-oligolactide-PEO-oligolactide- (meth)acrylate, polyethylene glycol) (PEO), polypropylene glycol) (PPO), PEO-PPO-PEO copolymers (Pluronic®), poly(phosphazene), poly(methacrylates), poly(N-vinylpyrrolidone), PL(G)A-PEO-PL(G)A copolymers, or polyethylene imine). In some embodiments, the hydrogel comprises agarose, polyvinylalcohol, poly(lactic-co-glycolic)acid (PLGA) polymers, (meth)acrylate-oligolactide-PEO- oligolactide-(meth)acrylate, poly(ethylene glycol) (PEO), polypropylene glycol) (PPO), PEO-PPO-PEO copolymers (Pluronic®), poly(phosphazene), poly(methacrylates), poly(N-vinylpyrrolidone), PL(G)A- PEO-PL(G)A copolymers, or poly(ethylene imine), or any combinations thereof. In some embodiments, the hydrogel comprises agarose, polyvinylalcohol, poly(lactic-co-glycolic)acid (PLGA) polymers, (meth)acrylate-oligolactide-PEO-oligolactide-(meth)acrylate, poly(ethylene glycol) (PEO), polypropylene glycol) (PPO), PEO-PPO-PEO copolymers (Pluronic®), poly(phosphazene), poly(methacrylates), poly(N-vinylpyrrolidone), PL(G)A-PEO-PL(G)A copolymers, or poly(ethylene imine). In some embodiments, the hydrogel comprises agarose, hyaluronans, chitosans, collagen, dextran, pectin, polylysine, polystyrene, gelatin, or starch, or any combinations thereof. In some embodiments, the hydrogel comprises agarose, hyaluronans, chitosans, collagen, dextran, pectin, polylysine, gelatin, or starch. In some embodiments, the hydrogel comprises agarose.

[00109] In some embodiments, an internal infrastructure described herein is porous. In some embodiments, the internal infrastructure comprises an outer surface charge. In some embodiments, the outer surface charge is charge potential. In some embodiments, the charge potential is positive. In some embodiments, the charge potential is negative. In some embodiments, the charge potential is neutral. In some embodiments, the surface charge comprises an ionic interaction or a polar covalent interaction, or any combinations thereof. In some embodiments, the surface charge comprises an ionic interaction or a polar covalent interaction. In some embodiments, the surface charge comprises a polar covalent interaction. In some embodiments, the surface charge comprises an ionic interaction. In some embodiments, the ionic interaction between the Zwitterionic lipids, the cationic lipids, or the anionic lipids, or any combinations thereof, and the surface of the internal infrastructure supports formation of the containment layer. In some embodiments, the ionic interaction between the Zwitterionic lipids or the anionic lipids, or any combinations thereof, and the surface of the internal infrastructure supports formation of the containment layer. In some embodiments, the ionic interaction between the Zwitterionic lipids or the cationic lipids, or any combinations thereof, and the surface of the internal infrastructure supports formation of the containment layer. In some embodiments, the ionic interaction between the cationic lipids or the anionic lipids, or any combinations thereof, and the surface of the internal infrastructure supports formation of the containment layer.

[00110] In some embodiments, an ionic interaction described herein is between the Zwitterionic lipids, the cationic lipids, or the anionic lipids and the surface of the internal infrastructure supports formation of the containment layer. In some embodiments, the ionic interaction between the Zwitterionic lipids or the anionic lipids and the surface of the internal infrastructure supports formation of the containment layer. In some embodiments, the ionic interaction between the Zwitterionic lipids or the cationic lipids and the surface of the internal infrastructure supports formation of the containment layer. In some embodiments, the ionic interaction between the cationic lipids or the anionic lipids and the surface of the internal infrastructure supports formation of the containment layer. In some embodiments, the containment layer is an extracellular containment layer.

[00111] In some embodiments, a polar covalent interaction described herein is between the Zwitterionic lipids, the cationic lipids, or the anionic lipids, or any combinations thereof, and the internal infrastructure of the containment layer further comprises an anchoring layer. In some embodiments, the polar covalent interaction is between the Zwitterionic lipids or the anionic lipids, or any combinations thereof, and the internal infrastructure of the containment layer further comprises an anchoring layer. In some embodiments, the polar covalent interaction is between the Zwitterionic lipids or the cationic lipids, or any combinations thereof, and the internal infrastructure of the containment layer further comprises an anchoring layer. In some embodiments, the polar covalent interaction is between the cationic lipids or the anionic lipids, or any combinations thereof, and the internal infrastructure of the containment layer further comprises an anchoring layer.

[00112] In some embodiments, a polar covalent interaction described herein is between the Zwitterionic lipids, the cationic lipids, or the anionic lipids, and the internal infrastructure of the containment layer further comprises an anchoring layer. In some embodiments, the polar covalent interaction is between the Zwitterionic lipids or the anionic lipids and the internal infrastructure of the containment layer further comprises an anchoring layer. In some embodiments, the polar covalent interaction is between the Zwitterionic lipids or the cationic lipids and the internal infrastructure of the containment layer further comprises an anchoring layer. In some embodiments, the polar covalent interaction is between the cationic lipids or the anionic lipids and the internal infrastructure of the containment layer further comprises an anchoring layer.

[00113] In some embodiments, an anchoring layer described herein is covalently bonded to the containment layer or the internal infrastructure, or any combinations thereof. In some embodiments, the anchoring layer is covalently bonded to the containment layer or the internal infrastructure. In some embodiments, the anchoring layer is covalently bonded to the containment layer. In some embodiments, the anchoring layer is covalently bonded to the internal infrastructure. In some embodiments, the anchoring layer comprises a hydrophobic anchoring element. In some embodiments, the hydrophobic anchoring element of the anchoring layer comprises a monomer, an oligomer, or a polymer, or any combinations thereof. In some embodiments, the hydrophobic anchoring element of the anchoring layer comprises an oligomer or a polymer, or any combinations thereof. In some embodiments, the hydrophobic anchoring element of the anchoring layer comprises a monomer. In some embodiments, the hydrophobic anchoring element of the anchoring layer comprises an oligomer. In some embodiments, the hydrophobic anchoring element of the anchoring layer comprises a polymer. In some embodiments, the anchoring layer comprises an alkyl chain or an alkenyl chain, or any combinations thereof. In some embodiments, the anchoring layer comprises an alkyl chain or an alkenyl chain. In some embodiments, the anchoring layer comprises an alkyl chain. In some embodiments, the anchoring layer comprises an alkenyl chain. In some embodiments, the anchoring layer comprises a Ci-is alkyl or a C2-18 alkenyl, or any combinations thereof, wherein each alkyl or alkenyl is optionally substituted with one or more substituents. In some embodiments, the anchoring layer comprises a Ci-is alkyl, wherein each alkyl is optionally substituted with one or more substituents. In some embodiments, the anchoring layer comprises a C2-18 alkenyl, wherein each alkenyl is optionally substituted with one or more substituents. In some embodiments, the anchoring layer comprises hexylamine, octyl amine, decyl chloride, dodecyl amine, or octadecyl isocyanate, or any combinations thereof. In some embodiments, the anchoring layer comprises decyl chloride, dodecyl amine, or octadecyl isocyanate, or any combinations thereof. In some embodiments, the anchoring layer comprises hexylamine, octyl amine, decyl chloride, or dodecyl amine, or any combinations thereof. In some embodiments, the anchoring layer comprises hexylamine. In some embodiments, the anchoring layer comprises octyl amine. In some embodiments, the anchoring layer comprises decyl chloride. In some embodiments, the anchoring layer comprises dodecyl amine. In some embodiments, the anchoring layer comprises octadecyl isocyanate.

[00114] In some embodiments, a plurality of intercellular router constructs described herein comprise different internal infrastructures. In some embodiments, the plurality of intercellular router constructs comprise at most two different internal infrastructures. In some embodiments, the plurality of intercellular router constructs comprise at most three different internal infrastructures. In some embodiments, the plurality of intercellular router constructs comprise at most four different internal infrastructures. In some embodiments, the plurality of intercellular router constructs comprise at least two different internal infrastructures. In some embodiments, the plurality of intercellular router constructs comprise at least three different internal infrastructures. In some embodiments, the plurality of intercellular router constructs comprise at least four different internal infrastructures. In some embodiments, the internal infrastructures comprise a hydrogel, a polystyrene, calcium carbonate, silicon oxide, calcium alginate, calcium phosphate, SiCfC 18. or lithium iron phosphate. In some embodiments, the internal infrastructures comprise a hydrogel or calcium phosphate. In some embodiments, the internal infrastructures comprise a hydrogel. In some embodiments, the internal infrastructures comprise calcium phosphate. In some embodiments, the internal infrastructures comprise a polystyrene. In some embodiments, the internal infrastructures comprise calcium carbonate. In some embodiments, the internal infrastructures comprise calcium alginate. In some embodiments, the internal infrastructures comprise silicon oxide. In some embodiments, the internal infrastructures comprise SiCfC 18. In some embodiments, the internal infrastructure comprises lithium iron phosphate.

[00115] In some embodiments, the intercellular router construct comprises a surface charge. In some embodiments, the intracellular router construct comprises a positive surface charge. In some embodiment, the intracellular router construct comprises a negative surface charge. In some embodiments, the intercellular router construct comprises a neutral surface charge. [00116] In some embodiments, a networking element described herein comprises an array of connections to transfer the cargo molecule between the intercellular router construct and the cell, between the cell and the intercellular router, between intercellular router constructs, or between cells, or any combinations thereof. In some embodiments, the networking element comprises an array of connections to transfer the cargo molecule between the intercellular router construct and the cell, between the cell and the intercellular router, or any combinations thereof. In some embodiments, the networking element comprises an array of connections to transfer the cargo molecule between the cell and the intercellular router, between intercellular router constructs, or between cells, or any combinations thereof. In some embodiments, the networking element comprises an array of connections to transfer the cargo molecule between the intercellular router construct and the cell. In some embodiments, the networking element comprises an array of connections to transfer the cargo molecule between the cell and the intercellular router. In some embodiments, the networking element comprises an array of connections to transfer the cargo molecule between intercellular router constructs. In some embodiments, the networking element comprises an array of connections to transfer the cargo molecule between the cell, between the intercellular router construct and the cell, between the cell and the intercellular router, or between intercellular router constructs, or any combinations thereof. In some embodiments, the networking element comprises an array of connections to transfer the cargo molecule between the cells.

[00117] In some embodiments, a networking element described herein facilitates an interaction between the intercellular router construct and the cell. In some embodiments, the networking element is more dense than water. In some embodiments, the networking element and the intercellular router construct taken together are more dense than water. In some embodiments, the networking element is more dense than water. In some embodiments, the intercellular router is more dense than water. In some embodiments, the networking element and the intercellular router construct further comprise a material that is more dense than water. In some embodiments, the material is miscible. In some embodiments, the material is miscible and the internal infrastructure of the intercellular router construct comprises a fluid body. In some embodiments, the material comprises a sugar, a protein, or a water soluble polymer, or any combinations thereof. In some embodiments, the material comprises a sugar, a protein, or a water soluble polymer. In some embodiments, the material comprises a protein or a water soluble polymer, or any combinations thereof. In some embodiments, the material comprises a protein or a water soluble polymer. In some embodiments, the material comprises a sugar or a protein, or any combinations thereof. In some embodiments, the material comprises a sugar or a protein. In some embodiments, the material comprises a sugar. In some embodiments, the material comprises a protein. In some embodiments, the material comprises a water soluble polymer.

[00118] In some embodiments, a sugar described herein is sucrose. In some embodiments, the sugar is about 100 mM to 1 M. In some embodiments, the sugar is about 150 mM to 1 M. In some embodiments, the sugar is about 200 mM to 1 M. In some embodiments, the sugar is about 250 mM to 1 M. In some embodiments, the sugar is about 300 mM to 1 M. In some embodiments, the sugar is about 350 mM to 1 M. In some embodiments, the sugar is about 400 mM to 1 M. In some embodiments, the sugar is about 450 mM to 1 M. In some embodiments, the sugar is about 500 mM to 1 M. In some embodiments, the sugar is about 600 mM to 1 M. In some embodiments, the sugar is about 700 mM to 1 M. In some embodiments, the sugar is about 800 mM to 1 M. In some embodiments, the sugar is about 900 mM to 1 M. In some embodiments, the sugar is about 100 pM to 1 M. In some embodiments, the sugar is about 150 pM to 1 M. In some embodiments, the sugar is about 200 pM to 1 M. In some embodiments, the sugar is about 250 pM to 1 M. In some embodiments, the sugar is about 300 pM to 1 M. In some embodiments, the sugar is about 350 pM to 1 M. In some embodiments, the sugar is about 400 pM to 1 M. In some embodiments, the sugar is about 450 pM to 1 M. In some embodiments, the sugar is about 500 pM to 1 M. In some embodiments, the sugar is about 550 pM to 1 M. In some embodiments, the sugar is about 600 pM to 1 M. In some embodiments, the sugar is at most 100 mM. In some embodiments, the sugar is at most 150 mM. In some embodiments, the sugar is at most 200 mM. In some embodiments, the sugar is at most 250 mM. In some embodiments, the sugar is at most 300 mM. In some embodiments, the sugar is at most 350 mM. In some embodiments, the sugar is at most 400 mM. In some embodiments, the sugar is ab at most 450 mM. In some embodiments, the sugar is at most 500 mM. In some embodiments, the sugar is at most 600 mM. In some embodiments, the sugar is at most 700 mM. In some embodiments, the sugar is at most 900 mM. In some embodiments, the sugar is at most 1 M. [00119] In some embodiments, an interaction between the intercellular router construct and the cell described herein comprises an electrostatic affinity or a hydrophobic affinity. In some embodiments, the interaction between the intercellular router construct and the cell comprises an electrostatic affinity. In some embodiments, the interaction between the intercellular router construct and the cell comprises a hydrophobic affinity. In some embodiments, the interaction between the intercellular router construct and the cell comprises an electrostatic affinity or a hydrophobic affinity between the containment layer and the cell. In some embodiments, the interaction between the intercellular router construct and the cell comprises an electrostatic affinity between the containment layer and the cell. In some embodiments, the interaction between the intercellular router construct and the cell comprises a hydrophobic affinity between the containment layer and the cell.

[00120] In some embodiments, a dopant lipid described herein comprises a cationic lipid. In some embodiments, a membrane dopant lipid comprises a cationic lipid. In some embodiments, the cationic lipid comprises dioleoyl phosphatidylethanolamine (DOPE), dioleoyl trimethylammonium-propane (DOTAP), or dioleyloxy-N-sperminecarboxamido ethyl -N,N-dimethyl-l-propanaminium (DOSPA), or any combination thereof. In some embodiments, the cationic lipid comprises dioleoyl phosphatidylethanolamine (DOPE), dioleoyl trimethylammonium-propane (DOTAP), or dioleyloxy-N- sperminecarboxamido ethyl-N,N-dimethyl-l-propanaminium (DOSPA). In some embodiments, the cationic lipid comprises dioleoyl trimethylammonium-propane (DOTAP) or dioleyloxy-N- sperminecarboxamido ethyl-N,N-dimethyl-l-propanaminium (DOSPA), or any combination thereof. In some embodiments, the cationic lipid comprises dioleoyl phosphatidylethanolamine (DOPE) or dioleoyl trimethylammonium-propane (DOTAP), or any combination thereof. In some embodiments, the cationic lipid comprises dioleoyl phosphatidylethanolamine (DOPE). In some embodiments, the cationic lipid comprises dioleoyl trimethylammonium -propane (DOTAP). In some embodiments, the cationic lipid comprises dioleyloxy-N-sperminecarboxamido ethyl-N,N-dimethyl-l-propanaminium (DOSPA). [00121] In some embodiments, the dopant lipid comprises an anionic lipid. In some embodiments, a membrane dopant lipid comprises an anionic lipid. In some embodiments, the anionic lipid comprises dioleoyl phosphatidylglycerol (DOPG) or dioleoyl phosphatidylserine (DOPS), or any combinations thereof. In some embodiments, the anionic lipid comprises dioleoyl phosphatidylglycerol (DOPG). In some embodiments, the anionic lipid comprises dioleoyl phosphatidylserine (DOPS).

[00122] In some embodiments, a dopant lipid described herein comprises a Zwitterionic lipid. In some embodiments, a membrane dopant lipid comprises a Zwitterionic lipid. In some embodiments, the Zwitterionic lipid comprises hydrogenated soy phosphatidylcholine (HSPC), soy phosphatidylcholine (Soy PC), or dioleoyl phosphatidylcholine (DOPC), or any combinations thereof. In some embodiments, the Zwitterionic lipid comprises hydrogenated soy phosphatidylcholine (HSPC) or dioleoyl phosphatidylcholine (DOPC), or any combinations thereof. In some embodiments, the Zwitterionic lipid comprises soy phosphatidylcholine (Soy PC) or dioleoyl phosphatidylcholine (DOPC), or any combinations thereof. In some embodiments, the Zwitterionic lipid comprises hydrogenated soy phosphatidylcholine (HSPC). In some embodiments, the Zwitterionic lipid comprises soy phosphatidylcholine (Soy PC). In some embodiments, the Zwitterionic lipid comprises hydrogenated dioleoyl phosphatidylcholine (DOPC).

[00123] In some embodiments, a dopant lipid described herein comprises a Zwitterionic lipids, a cationic lipids, or an anionic lipids, or any combinations thereof. In some embodiments, the dopant lipid comprises a Zwitterionic lipids, a cationic lipids, or an anionic lipids. In some embodiments, the dopant lipid comprises a Zwitterionic lipids or an anionic lipids, or any combinations thereof. In some embodiments, the dopant lipid comprises a Zwitterionic lipids or an anionic lipids. In some embodiments, the dopant lipid comprises a Zwitterionic lipids or a cationic lipids, or any combinations thereof. In some embodiments, the dopant lipid comprises a Zwitterionic lipids or a cationic lipids. In some embodiments, the dopant lipid comprises a cationic lipids or an anionic lipids, or any combinations thereof. In some embodiments, the dopant lipid comprises a cationic lipids or an anionic lipids.

[00124] In some embodiments, a membrane dopant lipid described herein comprises a Zwitterionic lipid, a cationic lipid, or an anionic lipid, or any combinations thereof. In some embodiments, the membrane dopant lipid comprises a Zwitterionic lipid, a cationic lipid, or an anionic lipid. In some embodiments, the membrane dopant lipid comprises a Zwitterionic lipid or an anionic lipid, or any combinations thereof. In some embodiments, the membrane dopant lipid comprises a Zwitterionic lipid or an anionic lipid. In some embodiments, the membrane dopant lipid comprises a Zwitterionic lipid or a cationic lipid, or any combinations thereof. In some embodiments, the membrane dopant lipid comprises a Zwitterionic lipid or a cationic lipid. In some embodiments, the membrane dopant lipid comprises a cationic lipid or an anionic lipid, or any combinations thereof. In some embodiments, the membrane dopant lipid comprises a cationic lipid or an anionic lipid. [00125] In some embodiments, an intercellular router construct described herein comprises a fusogenic element. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct comprises at most 5 mol%, 10 mol%, 15 mol%, 20 mol%, or 25 mol% of the dopant lipid. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct comprises at most 5 mol%, 10 mol%, 15 mol%, or 20 mol% of the dopant lipid. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct comprises at most 5 mol%, 10 mol%, or 15 mol% of the dopant lipid. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct comprises at most 5 mol% or 10 mol% of the dopant lipid. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct comprises at most 5 mol% of the dopant lipid. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct comprises at most 10 mol% of the dopant lipid. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct comprises at most 15 mol% of the dopant lipid. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct comprises at most 20 mol% of the dopant lipid. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct comprises at most 25 mol% of the dopant lipid.

[00126] In some embodiments, an intercellular router construct system described herein comprises a fusogenic element. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct system comprises at most 5 mol%, 10 mol%, 15 mol%, 20 mol%, or 25 mol% of the dopant lipid. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct system comprises at most 5 mol%, 10 mol%, 15 mol%, or 20 mol% of the dopant lipid. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct system comprises at most 5 mol%, 10 mol%, or 15 mol% of the dopant lipid. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct system comprises at most 5 mol% or 10 mol% of the dopant lipid. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct system comprises at most 5 mol% of the dopant lipid. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct system comprises at most 10 mol% of the dopant lipid. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct system comprises at most 15 mol% of the dopant lipid. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct system comprises at most 20 mol% of the dopant lipid. In some embodiments, the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router construct system comprises at most 25 mol% of the dopant lipid.

[00127] In some embodiments, a dopant lipid described herein comprises a cationic lipid. In some embodiments, the cationic lipid comprises dioleoyl phosphatidylethanolamine (DOPE), dioleoyl trimethylammonium-propane (DOTAP), or dioleyloxy-N-sperminecarboxamido ethyl-N,N-dimethyl-l- propanaminium (DOSPA), or any combination thereof. In some embodiments, the cationic lipid comprises dioleoyl phosphatidylethanolamine (DOPE) or dioleoyl trimethylammonium-propane (DOTAP), or any combination thereof. In some embodiments, the cationic lipid comprises dioleoyl trimethylammonium-propane (DOTAP) or dioleyloxy-N-sperminecarboxamido ethyl -N, N-dimethyl- 1- propanaminium (DOSPA), or any combination thereof. In some embodiments, the cationic lipid comprises dioleoyl phosphatidylethanolamine (DOPE). In some embodiments, the cationic lipid comprises dioleoyl trimethylammonium-propane (DOTAP). In some embodiments, the cationic lipid comprises dioleyloxy-N-sperminecarboxamido ethyl-N,N-dimethyl-l-propanaminium (DOSPA).

[00128] In some embodiments, a fusogenic element described herein comprises a peptide. In some embodiments, the peptide is present at a concentration of 10 nM to 5 mM. In some embodiments, the fusogenic element comprises a peptide. In some embodiments, the peptide is present at a concentration of 10 nM to 1 mM. In some embodiments, the peptide is present at a concentration of 50 nM to 1 mM. In some embodiments, the peptide is present at a concentration of 100 nM to 1 mM. In some embodiments, the peptide is present at a concentration of 150 nM to 1 mM. In some embodiments, the peptide is present at a concentration of 200 nM to 1 mM. In some embodiments, the peptide is present at a concentration of 250 nM to 1 mM. In some embodiments, the peptide is present at a concentration of 300 nM to 1 mM. In some embodiments, the peptide is present at a concentration of 350 nM to 1 mM. In some embodiments, the peptide is present at a concentration of 400 nM to 1 mM. In some embodiments, the peptide is present at a concentration of 450 nM to 1 mM. In some embodiments, the peptide is present at a concentration of 500 nM to 1 mM. In some embodiments, the peptide is present at a concentration of 10 pM to 1 mM. In some embodiments, the peptide is present at a concentration of 50 pM to 1 mM. In some embodiments, the peptide is present at a concentration of 100 pM to 1 mM. In some embodiments, the peptide is present at a concentration of 150 pM to 1 mM. In some embodiments, the peptide is present at a concentration of 200 pM to 1 mM. In some embodiments, the peptide is present at a concentration of 250 pM to 1 mM.In some embodiments, the peptide is present at a concentration of 300 pM to 1 mM. In some embodiments, the peptide is present at a concentration of 350 pM to 1 mM. In some embodiments, the peptide is present at a concentration of 400 pM to 1 mM. In some embodiments, the peptide is present at a concentration of 450 pM to 1 mM. In some embodiments, the peptide is present at a concentration of 500 pM to 1 mM. In some embodiments, the peptide is present at a concentration of at most 10 nM. In some embodiments, the peptide is present at a concentration of at most 50 nM. In some embodiments, the peptide is present at a concentration of at most 100 nM. In some embodiments, the peptide is present at a concentration of at most 150 nM. In some embodiments, the peptide is present at a concentration of at most 200 nM. In some embodiments, the peptide is present at a concentration of at most 250 nM. In some embodiments, the peptide is present at a concentration of at most 300 nM. In some embodiments, the peptide is present at a concentration of at most 350 nM. In some embodiments, the peptide is present at a concentration of at most 400 nM. In some embodiments, the peptide is present at a concentration of at most 450 nM. In some embodiments, the peptide is present at a concentration of at most 500 nM. In some embodiments, the peptide is present at a concentration of at most 600 nM. In some embodiments, the peptide is present at a concentration of at most 700 nM. In some embodiments, the peptide is present at a concentration of at most 800 nM. In some embodiments, the peptide is present at a concentration of at most 900 nM. In some embodiments, the peptide is present at a concentration of at most 1 mM. In some embodiments, the peptide is present at a concentration of at most 5 mM. In some embodiments, the peptide comprises a TAT cell-penetrating peptide or a KALA peptide, or any combination thereof. In some embodiments, the peptide comprises a TAT cell-penetrating peptide or a KALA peptide. In some embodiments, the peptide comprises a TAT cell-penetrating peptide. In some embodiments, the peptide comprises a KALA peptide.

[00129] In some embodiments, a fusogenic element described herein comprises a nanoparticle In some embodiments, the nanoparticle is present at a concentration of 10 nM to 5 mM. In some embodiments, the nanoparticle is present at a concentration of 10 nM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 50 nM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 100 nM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 150 nM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 200 nM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 250 nM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 300 nM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 350 nM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 400 nM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 450 nM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 500 nM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 10 pM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 50 pM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 100 pM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 150 pM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 200 pM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 250 pM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 300 pM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 350 pM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 400 pM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 450 pM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of 500 pM to 1 mM. In some embodiments, the nanoparticle is present at a concentration of at most 10 nM. In some embodiments, the nanoparticle is present at a concentration of at most 50 nM. In some embodiments, the nanoparticle is present at a concentration of at most 100 nM. In some embodiments, the nanoparticle is present at a concentration of at most 150 nM. In some embodiments, the nanoparticle is present at a concentration of at most 200 nM. In some embodiments, the nanoparticle is present at a concentration of at most 250 nM. In some embodiments, the nanoparticle is present at a concentration of at most 300 nM. In some embodiments, the nanoparticle is present at a concentration of at most 350 nM. In some embodiments, the nanoparticle is present at a concentration of at most 400 nM. In some embodiments, the nanoparticle is present at a concentration of at most 450 nM. In some embodiments, the nanoparticle is present at a concentration of at most 500 nM. In some embodiments, the nanoparticle is present at a concentration of at most 600 nM. In some embodiments, the nanoparticle is present at a concentration of at most 700 nM. In some embodiments, the nanoparticle is present at a concentration of at most 800 nM. In some embodiments, the nanoparticle is present at a concentration of at most 900 nM. In some embodiments, the nanoparticle is present at a concentration of at most 1 mM. In some embodiments, the nanoparticle is present at a concentration of at most 5 mM. In some embodiments, the nanoparticle is present at a concentration of at most 1 pM. In some embodiments, the nanoparticle is present at a concentration of at most 10 pM. In some embodiments, the nanoparticle is present at a concentration of at most 100 pM. In some embodiments, the nanoparticle is present at a concentration of at most 150 pM. In some embodiments, the nanoparticle is present at a concentration of at most 200 pM. In some embodiments, the nanoparticle is present at a concentration of at most 250 pM. In some embodiments, the nanoparticle is present at a concentration of at most 300 pM. In some embodiments, the nanoparticle is present at a concentration of at most 350 pM. In some embodiments, the nanoparticle is present at a concentration of at most 400 pM. In some embodiments, the nanoparticle is present at a concentration of at most 450 pM. In some embodiments, the nanoparticle is present at a concentration of at most 500 pM. In some embodiments, the nanoparticle is present at a concentration of at least 50 nM. In some embodiments, the nanoparticle is present at a concentration of at least 100 nM. In some embodiments, the nanoparticle is present at a concentration of at least 150 nM. In some embodiments, the nanoparticle is present at a concentration of at least 200 nM. In some embodiments, the nanoparticle is present at a concentration of at least 250 nM. In some embodiments, the nanoparticle is present at a concentration of at least 300 nM. In some embodiments, the nanoparticle is present at a concentration of at least 350 nM. In some embodiments, the nanoparticle is present at a concentration of at least 400 nM. In some embodiments, the nanoparticle is present at a concentration of at least 500 nM. In some embodiments, the nanoparticle is present at a concentration of at least 1 pM. In some embodiments, the nanoparticle is present at a concentration of at least 10 pM.

[00130] In some embodiments, a nanoparticle described herein comprises zinc oxide, silica oxide, or a lipoprotein particle, or any combination thereof. In some embodiments, the nanoparticle comprises zinc oxide, silica oxide, or a lipoprotein, or any combination thereof. In some embodiments, the nanoparticle comprises zinc oxide, silica oxide, or a lipoprotein particle. In some embodiments, the nanoparticle comprises zinc oxide, silica oxide, or a lipoprotein. In some embodiments, the nanoparticle comprises zinc oxide, or silica oxide, or any combination thereof. In some embodiments, the nanoparticle comprises silica oxide or a lipoprotein particle, or any combination thereof. In some embodiments, the nanoparticle comprises zinc oxide or a lipoprotein particle, or any combination thereof. In some embodiments, the nanoparticle comprises zinc oxide. In some embodiments, the nanoparticle comprises silica oxide. In some embodiments, the nanoparticle comprises a lipoprotein particle. In some embodiments, the nanoparticle comprises a lipoprotein.

[00131] In some embodiments, a fusogenic element described herein comprises a protein. In some embodiments, the protein is present at a concentration of 10 nM to 5 mM. In some embodiments, the protein is present at a concentration of 10 nM to 1 mM. In some embodiments, the protein is present at a concentration of 50 nM to 1 mM. In some embodiments, the protein is present at a concentration of 100 nM to 1 mM. In some embodiments, the protein is present at a concentration of 150 nM to 1 mM. In some embodiments, the protein is present at a concentration of 200 nM to 1 mM. In some embodiments, the protein is present at a concentration of 250 nM to 1 mM. In some embodiments, the protein is present at a concentration of 300 nM to 1 mM. In some embodiments, the protein is present at a concentration of 350 nM to 1 mM. In some embodiments, the protein is present at a concentration of 400 nM to 1 mM. In some embodiments, the protein is present at a concentration of 450 nM to 1 mM. In some embodiments, the protein is present at a concentration of 500 nM to 1 mM. In some embodiments, the protein is present at a concentration of at most 10 nM. In some embodiments, the protein is present at a concentration of at most 50 nM. In some embodiments, the protein is present at a concentration of at most 100 nM. In some embodiments, the protein is present at a concentration of at most 150 nM. In some embodiments, the protein is present at a concentration of at most 200 nM. In some embodiments, the protein is present at a concentration of at most 250 nM. In some embodiments, the protein is present at a concentration of at most 300 nM. In some embodiments, the protein is present at a concentration of at most 350 nM. In some embodiments, the protein is present at a concentration of at most 400 nM. In some embodiments, the protein is present at a concentration of at most 450 nM. In some embodiments, the protein is present at a concentration of at most 500 nM. In some embodiments, the protein is present at a concentration of at most 600 nM. In some embodiments, the protein is present at a concentration of at most 700 nM. In some embodiments, the protein is present at a concentration of at most 800 nM. In some embodiments, the protein is present at a concentration of at most 900 nM. In some embodiments, the protein is present at a concentration of at most 1 mM. In some embodiments, the protein is present at a concentration of at most 5 mM. In some embodiments, the protein is present at a concentration of 10 pM to 1 mM. In some embodiments, the protein is present at a concentration of 50 pM to 1 mM. In some embodiments, the protein is present at a concentration of 100 pM to 1 mM. In some embodiments, the protein is present at a concentration of 150 pM to 1 mM. In some embodiments, the protein is present at a concentration of 200 pM to 1 mM. In some embodiments, the protein is present at a concentration of 250 pM to 1 mM. In some embodiments, the protein is present at a concentration of 300 pM to 1 mM. In some embodiments, the protein is present at a concentration of 350 pM to 1 mM. In some embodiments, the protein is present at a concentration of 400 pM to 1 mM. In some embodiments, the protein is present at a concentration of 450 pM to 1 mM. In some embodiments, the protein is present at a concentration of 500 pM to 1 mM.

[00132] In some embodiments, a protein described herein comprises a SNARE coiled-coiled protein or a catechol protein, or any combinations thereof. In some embodiments, the protein comprises a SNARE coiled-coiled protein or a catechol protein. In some embodiments, the protein comprises a SNARE coiled-coiled protein. In some embodiments, the protein comprises a catechol protein.

[00133] In some embodiments, a fusogenic element described herein comprises a lipid, a peptide, a nanoparticle, or a protein, or any combinations thereof. In some embodiments, the fusogenic element comprises a lipid, a peptide, a nanoparticle, or a protein. In some embodiments, the fusogenic element comprises a peptide, a nanoparticle, or a protein, or any combinations thereof. In some embodiments, the fusogenic element comprises a lipid, a nanoparticle, or a protein, or any combinations thereof. In some embodiments, the fusogenic element comprises a lipid, a peptide, or a protein, or any combinations thereof. In some embodiments, the fusogenic element comprises a lipid, a peptide, or a nanoparticle, or any combinations thereof. In some embodiments, the fusogenic element comprises a lipid or a peptide, or any combinations thereof. In some embodiments, the fusogenic element comprises a lipid or a nanoparticle, or any combinations thereof. In some embodiments, the fusogenic element comprises a lipid or a protein, or any combinations thereof. In some embodiments, the fusogenic element comprises a peptide or a nanoparticle, or any combinations thereof. In some embodiments, the fusogenic element comprises a peptide or a protein, or any combinations thereof. In some embodiments, the fusogenic element comprises a nanoparticle or a protein, or any combinations thereof.

[00134] In some embodiments, an intercellular router construct described herein comprises a pore inducing element. In some embodiments, the pore inducing element comprises an exotoxin. In some embodiments, the exotoxin comprises a hemolysin. In some embodiments, the hemolysin is present at a concentration of 10 nM to 5 mM. In some embodiments, the hemolysin is present at a concentration of 10 nM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 50 nM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 100 nM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 150 nM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 200 nM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 250 nM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 300 nM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 350 nM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 400 nM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 450 nM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 500 nM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 10 pM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 50 pM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 100 pM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 150 pM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 200 pM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 250 pM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 300 pM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 350 pM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 400 pM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 450 pM to 1 mM. In some embodiments, the hemolysin is present at a concentration of 500 pM to 1 mM. In some embodiments, the hemolysin is present at a concentration of at most 10 nM. In some embodiments, the hemolysin is present at a concentration of at most 50 nM. In some embodiments, the hemolysin is present at a concentration of at most 100 nM. In some embodiments, the hemolysin is present at a concentration of at most 150 nM. In some embodiments, the hemolysin is present at a concentration of at most 200 nM. In some embodiments, the hemolysin is present at a concentration of at most 250 nM. In some embodiments, the hemolysin is present at a concentration of at most 300 nM. In some embodiments, the hemolysin is present at a concentration of at most 350 nM. In some embodiments, the hemolysin is present at a concentration of at most 400 nM. In some embodiments, the hemolysin is present at a concentration of at most 450 nM. In some embodiments, the hemolysin is present at a concentration of at most 500 nM. In some embodiments, the hemolysin is present at a concentration of at most 600 nM. In some embodiments, the hemolysin is present at a concentration of at most 700 nM. In some embodiments, the hemolysin is present at a concentration of at most 800 nM. In some embodiments, the hemolysin is present at a concentration of at most 900 nM. In some embodiments, the hemolysin is present at a concentration of at most 1 mM. In some embodiments, the hemolysin is present at a concentration of at most 5 mM.

[00135] In some embodiments, a hemolysin described herein comprises an alpha hemolysin (SLO), or a cholesterol-dependent cytolysin, or any combination thereof. In some embodiments, the hemolysin comprises an alpha hemolysin (SLO), or a cholesterol-dependent cytolysin. In some embodiments, the hemolysin comprises an alpha hemolysin (SLO). In some embodiments, the hemolysin comprises a cholesterol-dependent cytolysin.

[00136] In some embodiments, an intercellular router construct described herein comprises a local environment modulator. In some embodiments, the local environment modulator comprises a local dehydration reagent. In some embodiments, the local dehydration reagent comprises PEG-3000 or a Ca²⁺ salt, or any combination thereof. In some embodiments, the local dehydration reagent comprises PEG- 3000 or a Ca²⁺ salt. In some embodiments, the local dehydration reagent comprises PEG-3000. In some embodiments, the local dehydration reagent comprises a Ca²⁺ salt.

[00137] In some embodiments, intercellular router construct is from about 1 pM to about 10 mM in diameter. In some embodiments, intercellular router construct is from about 1 pM to about 1 mM in diameter. In some embodiments, intercellular router construct is from about 1 pM to about 50 pM in diameter. In some embodiments, intercellular router construct is from about 10 pM to about 30 pM in diameter. In some embodiments, intercellular router construct is from about 20 pM to about 25 pM in diameter.

[00138] In some embodiments, a networking element described herein comprises an array of membrane bound tubules connecting an intercellular router construct with a cell. In some embodiments, the array of membrane bound tubules are on the order of 100 nm to 1 micron in width. In some embodiments, the array of membrane bound tubules are on the order of 150 nm to 1 micron in width. In some embodiments, the array of membrane bound tubules are on the order of 200 nm to 1 micron in width. In some embodiments, the array of membrane bound tubules are on the order of 250 nm to 1 micron in width. In some embodiments, the array of membrane bound tubules are on the order of 300 nm to 1 micron in width. In some embodiments, the array of membrane bound tubules are on the order of 400 nm to 1 micron in width. In some embodiments, the array of membrane bound tubules are on the order of 500 nm to 1 micron in width. In some embodiments, the array of membrane bound tubules are on the order of 600 nm to 1 micron in width. In some embodiments, the array of membrane bound tubules are on the order of 700 nm to 1 micron in width. In some embodiments, the array of membrane bound tubules are on the order of 800 nm to 1 micron in width. In some embodiments, the array of membrane bound tubules are on the order of 50 nm to 1 micron in width. In some embodiments, the array of membrane bound tubules are on the order of at most 50 nm in width. In some embodiments, the array of membrane bound tubules are on the order of at most 100 nm in width. In some embodiments, the array of membrane bound tubules are on the order of at most 150 nm in width. In some embodiments, the array of membrane bound tubules are on the order of at most 200 nm in width. In some embodiments, the array of membrane bound tubules are on the order of at most 300 nm in width. In some embodiments, the array of membrane bound tubules are on the order of at most 400 nm in width. In some embodiments, the array of membrane bound tubules are on the order of at most 500 nm in width. In some embodiments, the array of membrane bound tubules are on the order of at most 600 nm in width. In some embodiments, the array of membrane bound tubules are on the order of at most 700 nm in width. In some embodiments, the array of membrane bound tubules are on the order of at most 800 nm in width. In some embodiments, the array of membrane bound tubules are on the order of at most 50 nm in width. In some embodiments, the array of membrane bound tubules are on the order of at most 1 micron in width.

[00139] In some embodiments, an array of membrane bound tubules described herein are on the order of 100 nm to 200 microns in length. In some embodiments, the array of membrane bound tubules are on the order of 50 nm to 200 microns in length. In some embodiments, the array of membrane bound tubules are on the order of 100 nm to 250 microns in length. In some embodiments, the array of membrane bound tubules are on the order of 50 nm to 250 microns in length. In some embodiments, the array of membrane bound tubules are on the order of 100 nm to 300 microns in length. In some embodiments, the array of membrane bound tubules are on the order of 150 nm to 200 microns in length. In some embodiments, the array of membrane bound tubules are on the order of 200 nm to 200 microns in length. In some embodiments, the array of membrane bound tubules are on the order of 250 nm to 200 microns in length. In some embodiments, the array of membrane bound tubules are on the order of 300 nm to 200 microns in length. In some embodiments, the array of membrane bound tubules are on the order of 350 nm to 200 microns in length. In some embodiments, the array of membrane bound tubules are on the order of 400 nm to 200 microns in length. In some embodiments, the array of membrane bound tubules are on the order of 450 nm to 200 microns in length. In some embodiments, the array of membrane bound tubules are on the order of 500 nm to 200 microns in length. In some embodiments, the array of membrane bound tubules are on the order of 600 nm to 200 microns in length. In some embodiments, the array of membrane bound tubules are on the order of 700 nm to 200 microns in length. In some embodiments, the array of membrane bound tubules are on the order of 800 nm to 200 microns in length. In some embodiments, the array of membrane bound tubules are on the order of at most 50 nm in length. In some embodiments, the array of membrane bound tubules are on the order of at most 100 nm in length. In some embodiments, the array of membrane bound tubules are on the order of at most 150 nm in length. In some embodiments, the array of membrane bound tubules are on the order of at most 200 nm in length. In some embodiments, the array of membrane bound tubules are on the order of at most 250 nm in length. In some embodiments, the array of membrane bound tubules are on the order of at most 300 nm in length. In some embodiments, the array of membrane bound tubules are on the order of at most 350 nm in length. In some embodiments, the array of membrane bound tubules are on the order of at most 400 nm in length. In some embodiments, the array of membrane bound tubules are on the order of at most 450 nm in length. In some embodiments, the array of membrane bound tubules are on the order of at most 500 nm in length. In some embodiments, the array of membrane bound tubules are on the order of at most 600 nm in length. In some embodiments, the array of membrane bound tubules are on the order of at most 700 nm in length. In some embodiments, the array of membrane bound tubules are on the order of at most 800 nm in length. In some embodiments, the array of membrane bound tubules are on the order of at most 200 microns in length. In some embodiments, the array of membrane bound tubules are on the order of at most 250 microns in length. In some embodiments, the array of membrane bound tubules are on the order of at most 300 microns in length.

[00140] In some embodiments, a cargo molecule described herein comprises a small molecule, a large molecule, a protein, a lipid, a nucleic acid, a metabolite, an ion, a nutrient, or an organelle, or any combinations thereof. In some embodiments, the cargo molecule comprises a small molecule, a large molecule, a protein, a lipid, a nucleic acid, a metabolite, an ion, a nutrient, or an organelle. In some embodiments, the cargo molecule comprises a small molecule, a large molecule, a protein, a lipid, a nucleic acid, a metabolite, or an organelle, or any combinations thereof. In some embodiments, the cargo molecule comprises a small molecule, a large molecule, a protein, a lipid, a nucleic acid, a metabolite, or an organelle. In some embodiments, the cargo molecule comprises a small molecule, a large molecule, a protein, a lipid, a nucleic acid, or a metabolite, or any combinations thereof. In some embodiments, the cargo molecule comprises a small molecule, a large molecule, a protein, a lipid, a nucleic acid, or a metabolite. In some embodiments, the cargo molecule comprises a protein, a lipid, a nucleic acid, or a metabolite, or any combinations thereof. In some embodiments, the cargo molecule comprises a protein, a lipid, a nucleic acid, or a metabolite. In some embodiments, the cargo molecule comprises a small molecule, a large molecule, a protein, or a nucleic acid, or any combinations. In some embodiments, the cargo molecule comprises a small molecule, a large molecule, a protein, or a nucleic acid. In some embodiments, the cargo molecule comprises a small molecule. In some embodiments, the cargo molecule comprises a large molecule. In some embodiments, the cargo molecule comprises a protein. In some embodiments, the cargo molecule comprises a lipid. In some embodiments, the cargo molecule comprises a nucleic acid. In some embodiments, the cargo molecule comprises a metabolite.

[00141] In some embodiments, a cargo molecule described herein comprises an exogenous material or a cell made material, or any combination thereof. In some embodiments, the cargo molecule comprises an exogenous material or a cell made material. In some embodiments, the cargo molecule comprises an exogenous material. In some embodiments, the cargo molecule comprises a cell made material. In some embodiments, the cargo molecule comprises cytosolic material, nuclear material, other organelle localizing material, or membrane bound material, or any combination thereof. In some embodiments, the cargo molecule comprises cytosolic material, nuclear material, other organelle localizing material, or membrane bound material. In some embodiments, the cargo molecule comprises cytosolic material or nuclear material, or any combination thereof. In some embodiments, the cargo molecule comprises cytosolic material. In some embodiments, the cargo molecule comprises nuclear material. In some embodiments, the cargo molecule comprises other organelle localizing material. In some embodiments, the cargo molecule comprises membrane bound material. In some embodiments, the nucleic acid comprises DNA or RNA. In some embodiments, the nucleic acid comprises DNA. In some embodiments, the nucleic acid comprises RNA. In some embodiments, the RNA comprises mRNA, miRNA, anti- miRNA, or siRNA, or any combinations thereof. In some embodiments, the RNA comprises mRNA, miRNA, anti-miRNA, or siRNA. In some embodiments, the RNA comprises miRNA, anti-miRNA, or siRNA, or any combinations thereof. In some embodiments, the RNA comprises miRNA, anti-miRNA, or siRNA. In some embodiments, the RNA comprises anti-miRNA or siRNA, or any combinations thereof. In some embodiments, the RNA comprises miRNA or siRNA, or any combinations thereof. In some embodiments, the RNA comprises mRNA or siRNA, or any combinations thereof. In some embodiments, the RNA comprises miRNA or anti-miRNA, or any combinations thereof. In some embodiments, the RNA comprises mRNA. In some embodiments, the RNA comprises miRNA. In some embodiments, the RNA comprises anti-miRNA. In some embodiments, the RNA comprises siRNA. In some embodiments, the RNA is fitc-mRNA.

[00142] In some embodiments, a protein described herein is a membrane receptor. In some embodiments, the membrane receptor is an immune antigen receptor. In some embodiments, the immune antigen receptor is a chimeric antigen receptor.

[00143] In one aspect, provided herein is a method of establishing an intercellular router construct system. In some embodiments, the method comprises contacting a cell with an intercellular router construct thereby creating a network that connects the cell and the containment layer and establishing an intercellular router construct system. In some embodiments, the intercellular network router construct comprises a containment layer comprising a membrane and an internal infrastructure. In some embodiments, the method comprises extending a networking from the cell to the intercellular router construct, which connects the cell and the containment layer. In some embodiments, the method further comprises extending a networking from the cell to a second cell, which connects the cell and the second cell. In some embodiments, the method comprises extending a plurality of networking from the cell. In some embodiments, the extending of the networking is stimulated by the presence of the intercellular router construct.

[00144] In another aspect, provided herein is a method of analyzing an intercellular router construct system (e.g., a tubule network). In some cases, the intercellular router construct system comprises a plurality of cells and/or a plurality of routers (e.g., beads). In some cases, the intercellular router construct system comprises at least one bead and at least one router (e.g., bead). The method of analyzing the intercellular router construct system may comprise graph theoretic analysis. For example, the method may comprise providing an image (e.g., a microscope image) of an intercellular router construct system (e.g., a tubule network), as depicted in FIG. 20A. This can be a fluorescence and/or brightfield microscope image. The method may comprise converting the image to a connection diagram, such as shown in FIG. 20B. In some cases, the connection diagram depicts one or more cells. In some cases, the connection diagram depicts one or more routers (e.g., beads). The connection diagram may further depict a connection (e.g., a tube or tubule) between a first cell and a second cell, between a first router and a second router, or between a cell and a router. In some cases, the connection diagram depicts a plurality of connections from a single cell or router (e.g., bead). In some cases, the connection diagram depicts the length of a connection (e.g., a tube or tubule). In some cases, the connection diagram depicts the angle of projection of a connection (e.g., a tube or tubule) from a cell or a router (e.g., bead).

[00145] In some cases, the connection diagram may be parsable by one or more graph theory algorithms. The one or more graph theory algorithms may calculate relevant statistics of the intercellular router construct system (e.g., a tubule network). In some cases, the analysis method may provide information about the topological properties of the intercellular router construct system. For example, the analysis method may provide information about the frequency of connections per bead in a tubule network, as shown in FIG. 21A, or the frequency of connections per cell in a tubule network, as shown in FIG. 21B. In another example, the analysis method may provide information about the frequency of lengths of connections in a tubule network, as shown in FIG. 22. In another example, the analysis method may provide information about the frequency of angle values of connections in a tubule network, as shown in FIG. 25.

[00146] In some cases, analyzing the intercellular router construct system provides bivariate statistics. For example, the analysis may provide information about the length of a connection (e.g., a bead to cell connection) vs. its angle of projection, as shown in FIG. 23. As another example, the analysis may provide information about the average length of connections for a specific bead or cell vs. the number of connections to that bead or cell, as shown in FIG. 24. As another example, the analysis may provide information about the average length per bead vs. span of angle per bead. These methods of analysis may be applied to compare different router constructs or applications. For example, these methods of analysis may be used to compare network evolution overtime. This can be applied for evaluation and optimization, for example, of different materials, compositions, or methods of generating an intercellular router construct provided herein.

[00147] In some embodiments, the cell is derived from the bone, breast, colon, esophagus, eye, heart, intestine, liver, lung, muscle, nervous system, oral cavity, nasal cavity, ovary, pancreas, skin, or stomach. In some embodiments, the cell is derived from the breast, colon, eye, liver, pancreas, or skin. In some embodiments, the cell is from the breast. In some embodiments, the cell is from the colon. In some embodiments, the cell is from the eye conjunctiva. In some embodiments, the cell is from the liver. In some embodiments, the cell is from the pancreas. In some embodiments, the cell is from the skin. In some embodiments, the cell is a fat cell. In some embodiments, the cell is a muscle cell. In some embodiments, the cell is from embryonic tissues. In some embodiments, the cell is from the nervous system.

[00148] In some embodiments, the cell is an ex-vivo modified cell (e.g., iPSC or CAR-T).

[00149] In some embodiments, the cell is a cancer cell.

[00150] In some embodiments, the cell comprises a tissue. In some embodiments, the tissue type is selected from adipose tissue, bone tissue, breast tissue, colon tissue, connective tissue, embryonic tissue, esophageal tissue, eye tissue, heart tissue, intestinal tissue, liver tissue, lung tissue, muscle tissue, nasal tissue, nervous tissue, oral tissue, ovarian tissue, pancreatic tissue, skin tissue, or stomach tissue. In some embodiment, the tissue is breast tissue. In some embodiments, the tissue is colon tissue. In some embodiments, the tissue is eye conjunctiva. In some embodiments, the tissue is liver tissue. In some embodiments, the tissue is lung tissue. In some embodiments, the tissue is ovary tissue. In some embodiments, the tissue is pancreatic tissue. In some embodiments, the tissue is skin tissue. In some embodiments, the tissue is adipose tissue. In some embodiments, the tissue is bone tissue. In some embodiments, the tissue is from the nervous system. In some embodiments, the tissue is nasal tissue. In some embodiments, the tissue is connective tissue. In some embodiments, the tissue is embryonic tissue. [00151] Ins some embodiments, the tissue is a scaffolded tissue, e.g., tissue ECM scaffold.

Lipids

[00152] In some embodiments, a containment layer described herein comprises a lipid or a dopant lipid. In some embodiments, the containment layer comprises a lipid. In some embodiments, the containment layer comprises a dopant lipid. In some embodiments, the containment layer comprises a cationic lipid. In some embodiments, the lipid is a cationic lipid. In some embodiments, the dopant lipid is a cationic lipid. In some embodiments, the containment layer comprises two or more cationic lipids. In some embodiments, the containment layer comprises two or more cationic dopant lipids. In some embodiments, the cationic lipid comprises a trimethyl sphingosine, a trimethyl phytosphingosine, or a pyridinium ceramide, or any combinations thereof. In some embodiments, the cationic lipid comprises l,2-di-O-octadecenyl-3-trimethylammonium propane chloride (DOTMA), l,2-dioleoyl-3- trimethylammonium-propane chloride (DOTAP), 1,2- dimyristoleoyl-sn-glycero-3-ethylphosplioclioline (14: 1) Nl-[2-((lS)-l-[(3-aminopropyl)amino]-4- [di(3-arrdno-propyl)amino]butylcarboxarrddo)ethyl]- 3,4-di[oleyloxy]-benzam (MVL5), Dioctadecylamido-glycylspermine (DOGS), 3b- [N-(N^AN'- dimcthylarrrinocthy I (carbamoyl | cholesterol (DC-Choi), Dioctadecyldimethylammonium Bromide (DDAB), a Saint lipid (e.g., SAINT-2, N- methyl-4-(dioleyl)methylpyridinium), 1,2-dimyristyloxypropyl- 3 -dimethylhydroxy ethylammonium bromide (DMRIE), l,2-dioleoyl-3 -dimethyl -hydroxyethyl ammonium bromide (DORIE), 1,2- dioleoyloxypropyl-3-dimethylhydroxyethyl ammonium chloride (DORI), Dioleyldimethylammonium chloride (DODAC), Dioctadecyldimethylammonium bromide (DDAB), 1,2- dilinoleyloxy-3- dimethylaminopropane (DLinDMA), 2,2-dilinoleyl-4-(2dimethylaminoethyl)-[l,3]- dioxolane (DLin- KC2-DMA), heptatriaconta-6,9,28,31-tetraen- 19-yl 4-(dimethylamino)butanoate (DLin-MC3 -DMA), l,2-Dioleoyloxy-3 -dimethylaminopropane (DODAP), l,2-Dioleyloxy-3- dimethylaminopropane (DODMA), Morpholinocholesterol (Mo-CHOL), (R)-5-(dimethylamino)pentane- 1,2-diyl dioleate hydrochloride (DODAPen-Cl), or (R)-N,N,N-trimethyl-4,5-bis(oleoyloxy)pentan-l- ammonium- chloride (DOTAPen), l,2-dioleoyl-3 -trimethylammonium -propane chloride (DOTAP), or any combinations thereof. Exemplary cationic lipids further comprise 3-(didodecylamino)- N1,N1,4- tridodecyl-l-piperazineethanamine (KL10), Nl-[2-(didodecylamino)ethyl]-Nl,N4,N4- tridodecyl- 1,4- piperazinediethanamine (KL22), 14,25-ditridecyl-15,18,21,24-tetraaza-octatriacontane (KL25), 1,2- dilinoleyloxy-N,N-dimethylaminopropane (DLin-DMA), 2,2-dilinoleyl-4- dimethylaminomethyl-[l,3]- dioxolane (DLin-K-DMA), heptatriaconta-6,9,28,31-tetraen- 19-yl 4- (dimethylamino)butanoate (DLin- MC3-DMA), 2,2-dilinoleyl-4-(2-dimethylaminoethyl)-[l,3]- dioxolane (DLin-KC2-DMA), 1,2- dioleyloxy-N,N-dimethylaminopropane (DODMA), 2-({8-[(3[3)- cholest-5-en-3-yloxy]octyl}oxy)-N,N- dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-l-yloxy]propan-l- amine (Octyl-CLinDMA), (2R)-2-({8- [(3P)-cholest-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3- [(9Z,12Z)-octadeca-9,12-dien-l-yloxy]propan- 1-amine (Octyl-CLinDMA (2R)), and (2S)-2-({8- [(3[3)-cholest-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-

3-[(9Z,12Z)-octadeca-9,12-dien-l- yloxy] propan- 1 -amine (Octyl-CLinDMA (2S)).

[00153] In some embodiments, a containment layer described herein comprises an anionic lipid. In some embodiments, the lipid is an anionic lipid. In some embodiments, the dopant lipid is an anionic lipid. In some embodiments, the containment layer comprises two or more anionic lipids. In some embodiments, the containment layer comprises two or more anionic dopant lipids. In some embodiments, the anionic lipid comprises an anionic sphingosine, an anionic phospholipid, an anionic phosphatidylinositol, an anionic inositol phosphate, an anionic cardiolipin, an anionic bis(monoacylglycero)phosphate, an anionic detergent that is not a sphingolipid or an anionic phospholipid, an anionic liponucleotide, or an anionic diacylglycerol pyrophosphate, or any combinations thereof. In some embodiments, the anionic lipid a phospholipid. In some embodiments, the anionic lipid comprises Dioleoylglycerolhemiglutarate (DOGG), Dimyristoylglycerolhemiglutarate (DMGG), Dioleoylglycerolhemiadipate (DOGA), Dimyristoylglycerolhemiadipate (DMGA), 4-{(l,2-Dioleoyl- ethyl)amino}-4-oxobutanoic acid (DOAS), 4-{( l,2-Dioleoyl-ethyl)amino}-4-oxopropanoic acid (DOAM), 4-{(l,2-Dioleoyl-ethyl)amino}-4-oxopentanoic acid (DOAG), 4-{(l,2-Dioleoyl-ethyl)amino}-

4-oxohexanoic acid (DOAA), 4-{(l,2-Dimyristoyl-ethyl)amino}-4-oxobutanoic acid (DMAS), 4-{(l,2- Dimyristoyl-ethyl)amino}-4-oxopropanoic acid (DMAM), 4-{(l,2-Dimyristoyl-ethyl)amino}-4- oxopentanoic acid (DMAG), 4-{(l,2-Dimyristoyl-ethyl) amino }-4-oxohexanoic acid (DMAA), 5,6- Dioleoyl-hexanoic acid (DOS), 4,5-Dioleoyl-pentanoic acid (DOM), 6,7-Dioleoyl-heptanoic acid (DOG), 7,8-Dioleoyl-octanoic acid (DOA), 5,6-Dimyristoyl-hexanoic acid (DMS), 4,5 -Dimyristoyl -pentanoic acid (DMM), 6,7-Dimyristoyl-heptanoic acid (DMG), 7,8-Dioleoyl-octanoic acid (DMA), cholesteryl hemisuccinate (CHEMS), Cholesterolhemimalonate (Chol-C3), Cholesterolhemiglutarate (Chol-C5), Cholesterolhemiadipate (Chol-C6), phosphatidylserine, palmitoylhomoserine, -tocopherol hemisuccinate (THS), 1,2-dierucoyl phosphatidylglycerol (DEPG), 1,2-dilauroyl phosphatidylglycerol (DLPG), 1,2- dimyristoyl phosphatidylglycerol (DMPG), 1,2-dioleoyl phosphatidylglycerol (DOPG), 1,2-dipalmitoyl phosphatidylglycerol (DPPS), 1,2-distearoyl phosphatidylglycerol (DSPG), 1 -palmitoyl -2 -oleoyl phosphatidylglycerol (POPG), or egg phosphatidylglycerol (EPG), or any combinations thereof. [00154] In some embodiments, a containment layer described herein comprises a Zwitterionic lipid. In some embodiments, the lipid is a Zwitterionic lipid. In some embodiments, the dopant lipid is a Zwitterionic lipid. In some embodiments, the containment layer comprises two or more Zwitterionic lipids. In some embodiments, the containment layer comprises two or more dopant Zwitterionic lipids. In some embodiments, the Zwitterionic lipid comprises Zwitterionic phosphatidylcholines, Zwitterionic phosphatidylethanolamines, Zwitterionic phosphatidylserines, Zwitterionic platelet activating factor phospholipids, Zwitterionic ether phospholipids, Zwitterionic plasmalogens, Zwitterionic oxidized phospholipids, Zwitterionic phospholipids for supported monolayers, Zwitterionic sterol modified phospholipids, or a Zwitterionic detergent that is not a sphingolipid or a phospholipid, or any combinations thereof. In some embodiments, the Zwitterionic lipid comprises 1,2-didecanoyl-sn-glycero- 3 -phosphocholine (DDPC), l,2-dierucoyl-sn-glycero-3-phosphocholine (DEPC), 1,2-dilinoleoyl-sn- glycero-3 -phosphocholine (DLOPC), l,2-dilauroyl-sn-glycero-3 -phosphocholine (DLPC), 1,2- dimyristoyl-sn-glycero-3-phosphocholine (DMPC), l,2-dioleoyl-sn-glycero-3 -phosphocholine (DOPC), l,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), l-myristoyl-2-palmitoyl-sn-glycero 3- phosphocholine (MPPC), 1 -myristoyl -2-stearoyl-sn-glycero-3 -phosphocholine (MSPC), 1 -palmitoyl -2- myristoyl-sn-glycero-3 -phosphocholine (PMPC), 1 -palmitoyl -2 -oleoyl-sn-glycero-3 -phosphocholine (POPC), 1 -palmitoyl -2-stearoyl-sn-glycero-3 -phosphocholine (PSPC), 1 -stearoyl -2 -myristoyl-sn- glycero-3 -phosphocholine (SMPC), 1 -stearoyl -2 -oleoyl-sn-glycero-3 -phosphocholine (SOPC), 1- stearoyl-2-palmitoyl-sn-glycero-3 -phosphocholine (SPPC), or egg phosphatidylcholine (EPC), or any combinations thereof.

[00155] In some embodiments, a containment layer described herein comprises cationic lipids, anionic lipids, or Zwitterionic lipids, or any combinations thereof. In some embodiments, the containment layer comprises cationic lipids, anionic lipids, Zwitterionic lipids, or dopant lipids, or any combinations thereof. In some embodiments, the containment layer comprises cationic lipids, anionic lipids, Zwitterionic lipids, or dopant lipids. In some embodiments, the containment layer comprises cationic lipids or Zwitterionic lipids, or any combinations thereof. In some embodiments, the containment layer comprises cationic lipids, Zwitterionic lipids, or dopant lipids, or any combinations thereof. In some embodiments, the containment layer comprises anionic lipids or Zwitterionic lipids, or any combinations thereof. In some embodiments, the containment layer comprises anionic lipids, Zwitterionic lipids, or dopant lipids, or any combinations thereof. In some embodiments, the containment layer comprises cationic lipids or anionic lipids, or any combinations thereof. In some embodiments, the containment layer comprises cationic lipids, anionic lipids, or dopant lipids, or any combinations thereof.

FURTHER EMBODIMENTS

[00156] Clause 1 : A therapeutic router construct system which has elements for (i) containment layer of desired molecules, (ii) internal infrastructure and support and (iii) networking to and/or between the cells and/or other routers for (iv) the purpose of exchanging of desired molecules across the established network while itself remaining extracellular.

[00157] Clause 2: The therapeutic router construct system of clause 1, wherein the containment layer of the therapeutic router insulates it from the environment and prevents the leakage, loss and/or exposure of desired molecules into the extracellular space.

[00158] Clause 3 : The therapeutic router construct system of clause 2, wherein the containment layer of the therapeutic router utilizes lipids.

[00159] Clause 4: The therapeutic router construct system of clause 3, wherein the lipids can vary in the head group charge as Zwitterionic, cationic or anionic.

[00160] Clause 5: The therapeutic router construct system of clause 3, wherein the lipids are majority (>50 mol%) Zwitterionic.

[00161] Clause 6: The therapeutic router construct system of clause 5, wherein the lipids are predominately zwitterionic (> 75 mol%) with some potential fraction (<25 mol%) of dopant lipids.

[00162] Clause 7: The therapeutic router construct system of clause 4, wherein the lipids are majority (>50 mol%) cationic.

[00163] Clause 8: The therapeutic router construct system of clause 7, wherein the lipids are predominately cationic (>75 mol%) with some potential fraction of dopant lipids (<25 mol%).

[00164] Clause 9: The therapeutic router construct system of clause 7, wherein the lipids are majority anionic.

[00165] Clause 10: The therapeutic router construct system of clause 7, wherein the lipids are predominately anionic (>75 mol%) with some potential fraction of dopant lipids (<25 mol%).

[00166] Clause 11 : The therapeutic router construct system of clause 4, wherein the lipids are a comparable (between 25 mol% and 75 mol%) mixture of Zwitterionic, cationic and/or anionic lipids.

[00167] Clause 12: The therapeutic router construct system of clause 7, 8 or 11, wherein the lipids are singly charged, doubly charged, triply charged or otherwise polycationic.

[00168] Clause 13: The therapeutic router construct system of clause 9, 10 and 11, wherein the lipids are singly charged, doubly charged, triply charged or otherwise polyanionic.

[00169] Clause 14: The therapeutic router construct system of clause 3, wherein the lipids can vary in the area of their head group specification.

[00170] Clause 15: The therapeutic router construct system of clause 14, wherein the head group can range from 40 to 80 square Angstroms.

[00171] Clause 16: The therapeutic router construct system of clause 3, wherein the lipids can vary in shape.

[00172] Clause 17: The therapeutic router construct system of clause 16, wherein the lipid includes but is not limited to cylindrical, cone-shaped, and inverted cone-shaped.

[00173] Clause 18: The therapeutic router construct system of clause 3, wherein the lipids can vary in the type of head group. [00174] Clause 19: The therapeutic router construct system of clause 18, wherein the head group includes but is not limited to phospholipid, glycolipid and sterile head groups.

[00175] Clause 20: The therapeutic router construct system of clause 3, wherein the lipids can vary in additional properties like length of chain, number of tails or number of double bonds.

[00176] Clause 21: The therapeutic router construct system of clause 3, wherein the lipids can be a polymerized version of multiple lipids with variation in the aforementioned properties in Clauses 4, 14, 16, 18 and 20.

[00177] Clause 22: The therapeutic router construct system of clause 3, wherein the lipids can be organized as a bilayer or monolayer membrane.

[00178] Clause 23 : The therapeutic router construct system of clause 22, wherein the membrane can be unilamellar or multilamellar.

[00179] Clause 24: The therapeutic router construct system of any one of clauses 4, 14, 16, 18 and 20 or 21, wherein the lipids include but are not limited to soy phosphatidylcholine (Soy PC), dioleoyl phosphatidylcholine (DOPC), dioleoyl phosphatidylglycerol (DOPG), dioleoyl phosphatidylethanolamine (DOPE), dioleoyl trimethylammonium-propane (DOTAP), dioleyloxy-N- sperminecarboxamido ethyl-N,N-dimethyl-l-propanaminium (DOSPA), dipalmitoylphosphatidyl choline (DPPC), hydrogenated soy phosphatidylcholine (HSPC), heptatriacont tetraene dimethylamino butanoate (DLin-DMA) or any combination thereof.

[00180] Clause 25: The therapeutic router construct system of clause 3, wherein the containment layer of the therapeutic router utilizes polymers, metallic substrates, plastics, ceramics, or fatty acids.

[00181] Clause 26: The therapeutic router construct system of clause 2, wherein the containment layer utilizes any combination of materials in Clause 3 and 25.

[00182] Clause 27: The therapeutic router construct system of clause 1, wherein multiple types of therapeutic routers are used together, each type with different materials in Clauses 3 and 25 for the containment in Clause 2.

[00183] Clause 28: The therapeutic router construct system of clause 27, wherein the combination are cationic lipid contained therapeutic router with anionic lipid contained therapeutic routers.

[00184] Clause 29: The therapeutic router construct system of clause 1, wherein the infrastructure offers its scale, support and geometry which further influence the possible network configurations. [00185] Clause 30: The therapeutic router construct system of clause 29, wherein the geometry of the therapeutic router achieved by the infrastructure can be spheroidal, planar, tubular or irregular.

[00186] Clause 31 : The therapeutic router construct system of clause 29, wherein the scale of the therapeutic router achieved by infrastructure can range from 10 nm to 10 mm.

[00187] Clause 32: The therapeutic router construct system of clause 31, wherein the size is specifically > 500 nm.

[00188] Clause 33: The therapeutic router construct system of clause 29, wherein the phase of the infrastructure component can be altered for different properties. [00189] Clause 34: The therapeutic router construct system of clause 33, wherein the infrastructure has a solid body.

[00190] Clause 35: The therapeutic router construct system of clause 34, wherein the solid body is an insoluble solid.

[00191] Clause 36: The therapeutic router construct system of clause 35, wherein the insoluble solid is a salt.

[00192] Clause 37: The therapeutic router construct system of clause 36, wherein the salt is a metal salt. [00193] Clause 38: The therapeutic router construct system of clause 37, wherein the metal is a divalent cation.

[00194] Clause 39: The therapeutic router construct system of clause 38, wherein the metal is calcium or magnesium.

[00195] Clause 40: The therapeutic router construct system of clause 37, wherein the anion of the salt is a phosphate, carbonate or sulfate.

[00196] Clause 41: The therapeutic router construct system of clause 39 or 40, wherein the metal is calcium phosphate.

[00197] Clause 42: The therapeutic router construct system of clause 35, wherein the insoluble solid is a metal.

[00198] Clause 43: The therapeutic router construct system of clause 35, wherein the insoluble solid is a metal oxide.

[00199] Clause 44: The therapeutic router construct system of clause 43, wherein the metal oxide is silica oxide or zinc oxide.

[00200] Clause 45: The therapeutic router construct system of clause 35, wherein the insoluble solid is a solid polymer.

[00201] Clause 46: The therapeutic router construct system of any one of clauses 36, 42, 43 or 45, wherein a combination of materials Clauses 36, 42, 43 and 45 are used as the insoluble solid in Clause 35 for the solid body infrastructure in Clause 34.

[00202] Clause 47: The therapeutic router construct system of clause 33, wherein the infrastructure has a fluid body.

[00203] Clause 48: The therapeutic router construct system of clause 47, wherein the fluid is a physiological buffer.

[00204] Clause 49: The therapeutic router construct system of clause 48, wherein the physiological buffer contains added salt(s).

[00205] Clause 50: The therapeutic router construct system of clause 49, wherein the salt is NaCl.

[00206] Clause 51 : The therapeutic router construct system of clause 48, wherein the physiological buffer contains pH regulator.

[00207] Clause 52: The therapeutic router construct system of clause 51, wherein the pH regulator is phosphate. [00208] Clause 53: The therapeutic router construct system of clause 48, wherein the physiological buffer contains additional nutritional, salts, sugars, amino acids and vitamins.

[00209] Clause 54: The therapeutic router construct system of clause 48, wherein the physiological buffer with the components mentioned in Clauses 49, 51 and 53 is minimal essential media or equivalent cell supporting basal media.

[00210] Clause 55: The therapeutic router construct system of clause 47, wherein the fluid is an otherwise nontoxic buffer like water.

[00211] Clause 56: The therapeutic router construct system of clause 47, wherein the fluid is a combination of a physiological buffer in Clause 48 and an otherwise nontoxic buffer in Clause 55. [00212] Clause 57: The therapeutic router construct system of clause 33, wherein the infrastructure has a semisolid body.

[00213] Clause 58: The therapeutic router construct system of clause 57, wherein the semi-solid material can be cross-linked or not cross-linked.

[00214] Clause 59: The therapeutic router construct system of clause 58, wherein if there is crosslinking in Clause 58 of it can be done physically or chemically.

[00215] Clause 60: The therapeutic router construct system of clause 59, wherein the cross-linked material is a hydrogel.

[00216] Clause 61 : The therapeutic router construct system of clause 60, wherein the hydrogel species includes but is not limited to agarose, hyaluronans, chitosans, collagen, dextran, pectin, polylysine, gelatin, starch, polyvinylalcohol, poly(lactic-co-glycolic)acid (PLGA) polymers, (meth)acrylate- oligolactide-PEO-oligolactide-(meth)acrylate, poly(ethylene glycol) (PEO), polypropylene glycol) (PPO), PEO-PPO-PEO copolymers (Pluronic®), poly(phosphazene), poly(methacrylates), poly(N- vinylpyrrolidone), PL(G)A-PEO-PL(G)A copolymers, or poly(ethylene imine).

[00217] Clause 62: The therapeutic router construct system of clause 29, wherein the infrastructure is porous.

[00218] Clause 63: The therapeutic router construct system of clause 29, wherein the infrastructures have an outer surface charge that enables the formation of various lipid containment layer.

[00219] Clause 64: The therapeutic router construct system of clause 63, wherein the infrastructure surface is ionic.

[00220] Clause 65 : The therapeutic router construct system of clause 64, wherein if the ionic surface naturally restricts the mobility of lipids used to form the containment layer.

[00221] Clause 66: The therapeutic router construct system of clause 63, wherein the infrastructure surface is polar covalent.

[00222] Clause 67 : The therapeutic router construct system of clause 66, wherein the polar covalent surface has an added anchoring layer that is utilized to bond to the lipid containment layer.

[00223] Clause 68: The therapeutic router construct system of clause 67, wherein the anchoring is through covalent boding. [00224] Clause 69: The therapeutic router construct system of clause 67, wherein the covalent bonding elements are hydrophobic.

[00225] Clause 70: The therapeutic router construct system of clause 67, wherein the covalent bonding elements are to monomers, oligomers, polymers or any combination thereof.

[00226] Clause 71: The therapeutic router construct system of clause 68, wherein the covalent bonding element with properties in Clauses 69 and 70 are alkyl chains.

[00227] Clause 72: The therapeutic router construct system of clause 71, wherein the length of the alkyl chains ranges from 1 to 18 carbons.

[00228] Clause 73: The therapeutic router construct system of clause 71, wherein the species of alkyl chains includes but are not limited to hexylamine, octyl amine, decyl chloride, dodecyl amine and octadecyl isocyante or any combination thereof.

[00229] Clause 74: The therapeutic router construct system of any one of clauses 30, 31, 33, 62 or 63, wherein two or more materials within different properties for the therapeutic router can be combined. [00230] Clause 75: The therapeutic router construct system of clause 74, wherein the infrastructure combination in Clause 74 is a combination of a hydrogel in Clause 60 and calcium phosphate in Clause 41.

[00231] Clause 76: The therapeutic router construct system of clause 1, wherein the networking element of the therapeutic router help establish an array of connections to and/or between cells and/or other routers to facilitate transfer of desire molecules.

[00232] Clause 77: The therapeutic router construct system of clause 76, wherein the networking element promote the therapeutic router's contact with the cell.

[00233] Clause 78: The therapeutic router construct system of clause 77, wherein the networking element promote contact with the cell by making the therapeutic router heavier than water.

[00234] Clause 79: The therapeutic router construct system of clause 29, wherein the infrastructure material also achieves this role for networking in Clause 78, through already being heavier than water. [00235] Clause 80: The therapeutic router construct system of clause 78, wherein added materials achieve this role for networking by increasing the therapeutic router's density greater than water.

[00236] Clause 81 : The therapeutic router construct system of clause 47, wherein the therapeutic router has a fluid body infrastructure as in Clause 47 and a miscible material is added to increase density greater than water as in Clause 80.

[00237] Clause 82: The therapeutic router construct system of clause 81, wherein the added miscible material includes but is not limited to a sugar, a protein, or a water soluble polymer, or any combinations thereof.

[00238] Clause 83: The therapeutic router construct system of clause 82, wherein the sugar is sucrose. [00239] Clause 84: The therapeutic router construct system of clause 82, wherein the amount of sugar ranges from 100 mM to 1 M. [00240] Clause 85: The therapeutic router construct system of clause 77, wherein the network elements promote contact by giving it the tendency to home and/or attach to cells through properties like electrostatic affinity, hydrophobicity, and/or increasing surface area for interaction.

[00241] Clause 86: The therapeutic router construct system of clause 29, wherein the infrastructure material in Clause 29 also achieves this role for networking in Clause 85, through already having this tendency to home and/or attach to cells.

[00242] Clause 87: The therapeutic router construct system of clause 41, wherein calcium phosphate infrastructure in Clause 41 achieves this role for networking in Clause 86.

[00243] Clause 88: The therapeutic router construct system of clause 2, wherein the containment material also achieve this role for networking in Clause 85, through already having this tendency to home and/or attach to cells.

[00244] Clause 89: The therapeutic router construct system of clause 7, wherein a cationic lipid containment achieves this role for networking.

[00245] Clause 90: The therapeutic router construct system of clause 85, wherein added materials achieve this role for networking by increasing or imbuing this tendency to home and/or attach to cells. [00246] Clause 91 : The therapeutic router construct system of clause 90, wherein the added materials are additional lipids doped (<25 mol %) into the containment layer.

[00247] Clause 92: The therapeutic router construct system of clause 91, wherein the dopant lipids are cationic with the properties listed in Clauses 12 and 14 through 23.

[00248] Clause 93 : The therapeutic router construct system of clause 92, wherein the species of cationic dopant lipid include but are not limited to dioleoyl phosphatidylethanolamine (DOPE), dioleoyl trimethylammonium -propane (DOTAP), and dioleyloxy-N-sperminecarboxamido ethyl -N, N-dimethyl- 1- propanaminium (DOSPA) or any combination thereof.

[00249] Clause 94: The therapeutic router construct system of clause 91, wherein the dopant lipids are anionic with the properties listed in Clauses 13 through 23.

[00250] Clause 95 : The therapeutic router construct system of clause 94, wherein the species of anionic dopant lipids include but are not limited to dioleoyl phosphatidylglycerol (DOPG) and dioleoyl phosphatidylserine (DOPS) or any combination thereof.

[00251] Clause 96: The therapeutic router construct system of clause 91, wherein the dopant lipids are zwitterionic with the properties listed in Clauses 14 through 23.

[00252] Clause 97 : The therapeutic router construct system of clause 96, wherein the species of zwitterionic dopant lipid include but are not limited to hydrogenated soy phosphatidylcholine (HSPC), soy phosphatidylcholine (Soy PC) and dioleoyl phosphatidylcholine (DOPC) or any combination thereof. [00253] Clause 98: The therapeutic router construct system of clause 91, wherein the dopant lipids are a combination of cationic, anionic and/or zwitterionic lipids in Clause 92, 94 and 96.

[00254] Clause 99: A therapeutic router construct system, wherein a combination of infrastructure materials in Clause 86, containment materials in Clause 88 and/or added materials in Clause 90 give the therapeutic router a greater tendency to home and/or attach to the cells as in Clause 85. [00255] Clause 100: The therapeutic router construct system of clause 77, wherein contact with the cell is promoted by networking for both making the therapeutic router heavier than water as in Clause 78 and giving it the tendency to home and/or attach to cells as in Clause 85.

[00256] Clause 101: The therapeutic router construct system of clause 76, wherein the networking element promote the therapeutic router's interaction with the cell membrane.

[00257] Clause 102: The therapeutic router construct system of clause 101, wherein the cell membrane interaction is mediated by a fusogenic element.

[00258] Clause 103: The therapeutic router construct system of clause 102, wherein the element is a dopant (<25 mol %) lipid in the containment layer.

[00259] Clause 104: The therapeutic router construct system of clause 103, wherein the dopant lipids are cationic with the properties listed in Clauses 12 and 14 through 23.

[00260] Clause 105: The therapeutic router construct system of clause 104, wherein the lipids include but are not limited to dioleoyl phosphatidylethanolamine (DOPE), dioleoyl trimethylammonium-propane (DOTAP), and dioleyloxy-N-sperminecarboxamido ethyl-N,N-dimethyl-l-propanaminium (DOSPA) or any combination thereof.

[00261] Clause 106: The therapeutic router construct system of clause 102, wherein the element is a peptide.

[00262] Clause 107: The therapeutic router construct system of clause 106, wherein the concentration of the peptide ranges from 10 nM to 1 mM.

[00263] Clause 108: The therapeutic router construct system of clause 106, wherein the peptides include but are not limited to the TAT cell-penetrating peptide and the KALA peptide or any combination thereof.

[00264] Clause 109: The therapeutic router construct system of clause 102, wherein the element is a nanoparticle.

[00265] Clause 110: The therapeutic router construct system of clause 109, wherein the concentration of the nanoparticle ranges from 10 nM to 1 mM.

[00266] Clause 111: The therapeutic router construct system of clause 109, wherein the nanoparticles include but is not limited to zinc oxide, silica oxide, and lipoprotein particles or any combination thereof. [00267] Clause 112: The therapeutic router construct system of clause 102, wherein the element is a protein.

[00268] Clause 113: The therapeutic router construct system of clause 112, wherein in the concentration ranges from 10 nM to 1 mM.

[00269] Clause 114: The therapeutic router construct system of clause 112, wherein the protein includes but is not limited to the SNARE coiled-coiled protein and the catechol protein.

[00270] Clause 115: The therapeutic router construct system of clause 102, wherein the element is a combination of materials in Clause 103, 106, 109 and 112.

[00271] Clause 116: The therapeutic router construct system of clause 102, wherein the cell membrane interaction is mediated by a pore inducing element. [00272] Clause 117: The therapeutic router construct system of clause 116, wherein the element is an exotoxin.

[00273] Clause 118: The therapeutic router construct system of clause 117, wherein the exotoxin is a hemolysin.

[00274] Clause 119: The therapeutic router construct system of clause 118, wherein in the concentration of the hemolysin ranges from 10 nM to 1 mM.

[00275] Clause 120: The therapeutic router construct system of clause 118, wherein the hemolysins include but are not limited to alpha hemolysin (SLO), cholesterol -dependent cytolysin or any combination thereof.

[00276] Clause 121: The therapeutic router construct system of clause 101, wherein the cell membrane interaction is mediated by a local environment modulator.

[00277] Clause 122: The therapeutic router construct system of clause 121, wherein the modulation is local dehydration.

[00278] Clause 123: The therapeutic router construct system of clause 122, wherein the local dehydrators include but are not limited to PEG-3000, Ca²⁺ or any combination thereof.

[00279] Clause 124: A therapeutic router construct system, wherein the therapeutic router's interaction with the cell membrane is promoted by a combination of fusogenic element in Clause 102, pore inducing element in Clause 116, and/or local environment modulator in Clause 121.

[00280] Clause 125: A therapeutic router construct system, wherein the networking element of the therapeutic router include a combination of elements to promote contact with the cell in Clause 77 and interaction with the cell membrane Clause 101.

[00281] Clause 126: A therapeutic router construct system of any one of the preceding clause, wherein a network is established between an extracellular construct and cells.

[00282] Clause 127: The therapeutic router construct system of clause 126, wherein construct is a therapeutic router in Clause 1.

[00283] Clause 128: The therapeutic router construct system of clause 126, wherein the network involves the induction and/or utilization of membrane bound tubule arrays to and/or between the extracellular construct and cells.

[00284] Clause 129: The therapeutic router construct system of clause 128, wherein the tubules are on the order of 100 nm to 1 micron in width.

[00285] Clause 130: The therapeutic router construct system of clause 128, wherein the tubules are on the order of 100 nm to 200 microns in length.

[00286] Clause 131: The therapeutic router construct system of clause 126, wherein the network is used to exchange materials.

[00287] Clause 132: The therapeutic router construct system of clause 131, wherein the types of materials exchanged across the network include but are not limited to small molecules, large molecules, peptides, proteins, lipids, nucleic acids, metabolites, ions, nutrients, organelles, other therapeutic molecules and any combination thereof. [00288] Clause 133: The therapeutic router construct system of clause 132, wherein the exchanged materials can be exogenous, cell made or any combination thereof.

[00289] Clause 134: The therapeutic router construct system of clause 132, wherein the exchanged materials can be cytosolic, nuclear or other organelle localizing, membrane bound or any combination thereof.

[00290] Clause 135: The therapeutic router construct system of clause 132, wherein the nucleic acids include DNA or RNA.

[00291] Clause 136: The therapeutic router construct system of clause 135, wherein the RNAs include but are not limited to mRNAs, miRNAs, anti-miRNAs, siRNAs or any combination thereof.

[00292] Clause 137: The therapeutic router construct system of clause 132, wherein the protein is a membrane receptor.

[00293] Clause 138: The therapeutic router construct system of clause 137, wherein the membrane receptor is an immune antigen receptor.

[00294] Clause 139: The therapeutic router construct system of clause 138, wherein the immune antigen receptor is a chimeric antigen receptor.

EXAMPLES

Example 1. Preparation of lipid-coated calcium phosphate beads

[00295] Lipid-coated calcium phosphate (lipoCaP) beads (FIG. 5) were prepared by weighing approximately 1 mg of CaP, Calcium Hydroxyapatite, CHT-I, 20pm product # 158-2000 (Bio-Rad Laboratories, Hercules, CA) in a 2 mL glass vial. Then 50 pL of lipids dissolved in chloroform at a concentration and composition of 1 mg/mL DOPC:DOTAP (90: 10 mol %) were added. Residual chloroform was removed with a SpeedVac™ SPD120 Vacuum Concentrator, (Thermo Fisher, Waltham, MA). After evaporation, the final lipoCaP bead solution was prepared by hydrating the beads in 500 pL of ultrapure water.

Example 2. Preparation of giant unilamellar vesicles

[00296] Giant unilamellar vesicles (GUVs) were prepared by depositing 10 pL of lipids dissolved in chloroform at a concentration and composition of 1 mg/mL DOPCDOPG (75:25 mol %) (FIG. 6) onto an approximately 15 mm diameter cutout disk of tracing paper (Bachmore 9”xl2” Artist’s Tracing Paper Pad). Residual chloroform was removed by placing the lipid-coated tracing paper into a speed vacuum for 20 minutes. Then the paper was hydrated in a 48 well plate in 150 pL of the desired cargo molecule along with 100 mM sucrose. The lipids were allowed to incubate for 2 hours and then the GUVs from the surface of the paper were harvested by washing the paper with a pipette with the tip cut off to minimize excessive shear forces. The GUVs were then stored in an Eppendorf tube until use.

Example 3. Preparation of agarose gel flakes [00297] Agarose gel flakes were prepared by mixing 8.5 g of mannitol (MilliporeSigma, Burlington, MA), 0.2 g of agarose (Thermo Fisher) and 20mLof water in a 100 mL round flask (14/20 neck). The mixture was microwaved until all of the ingredients were completely dissolved. Then the agarose solution was frozen in a dry ice /acetone mixture at 4-10 pascal for 24 hours. Next the agarose was rehydrated to form a gel with the addition of 40 mL of water. The gel was centrifuged, the supernatant was removed, and then the gel was rehydrating in water. The sequence was repeated twice with acetone and then the acetone was rinsed out using a 100 micron filter. The resulting slurry is kept in a sealed tube.

Example 4. Preparation of lipid-coated agarose gel flakes

[00298] Lipid-coated agarose gel (lipogel) flakes (FIG. 7) were prepared by dehydrating agarose gel flakes by mixing approximately 1 mg (around 1 mL volume of agarose in a 2 mL tube) in 1 mL acetone for 1 minute. The flakes were rinsed 2 times in acetone using a 40 pm nylon filter membrane. Then approximately 1.5 mL of a 0.1 mg/mL octadecyl isocyanate solution (diluted in acetone) was added and the reaction mixture was allowed to proceed for 3 hours. Next the octadecyl solution was rinsed 3 times in acetone using a 40 pm nylon filter. The resulting Cl 8 functionalized gel from the nylon filter was scraped and store in a 2 mL Eppendorf tube. Around 0.1 mg of the gel was incubate in 20 pL of 1 mg/mL DOPC in chloroform and 20 pL chloroform. Residual chloroform was removed with a speed vacuum . After evaporation, the final lipogel solution was prepared by rehydrating the lipogels in any aqueous solution with the desired cargo molecule.

Example 5. Surface charge of routers

[00299] The surface charge was measured in the lipid-coated calcium phosphate routers and the uncoated calcium phosphate starting material. The measurements were implemented with ZEN2600, MALVERN Industries limited. The results are shown in Table 1.

Table 1.

Example 6. pH determination

[00300] The effects of pH on two different suspensions containing lipid-coated calcium phosphate routers and uncoated calcium phosphate. The results are shown in Table 2.

Table 2.

Example 7. Preparation of cells

[00301] A breast cancer cell line, MDA-MB-231, was thawed from cryostorage at -80 °C. A volume of 1 mL of the cells was mixed with 10 mL of complete media (DMEM with glutaMAX, 10% FBS and 50 U/mL Penicillin-Streptomycin). The cells were centrifuged for 5 minutes at 2000 RPM, and the supernatant was removed leaving behind around 500 pL of solution. Then 6 mL of complete media was added. The cells were mixed and then add a T25 flask. The cells were allowed to grow for 3 days with the media (6 mL) being exchanged every 2 to 3 days. After the first passage when the cells were more than 50% confluent, the cells were split. About 3 mL of the cells were removed for cry opreservation (6 tubes with 500 pL cells and 500 pL cryomedia), 1 mL of the cells were moved to wells in a 48 well plate, and 2 mL of the cells were placed in another T25 flask. Once the cells in the T25 flask are confluent, in the second passage, the cells were split. Around 1 mL of the cells were removed for cry opreservation, 2 mL were removed for experiments, and 3 mL remained in the T25 flask.

Example 8 Cargo transfer from beads to cells

[00302] Following the methods to prepare the lipoCaP beads, the beads were hydrated with 500 pL of a desired cargo molecule diluted in ultrapure water. Various cargos have loaded into the beads including i) FITC-Dextran 3000 (Dextran 3000 g/mol with Fluorescein conjugated, (Thermo Fisher Scientific) at a concentration of 1 mM (FIG. 18 a,b), ii) siRNA at a concentration of 10 pM (FIG. 18 c,d), iii) anti -mi RNA at a concentration of 10 pM (FIG. 18 e,f), iv) eGFP mRNA at a concentration of 10 pM (FIG. 18 g,h).

[00303] A 48 well plate with around 60% confluent MDA-MB-231 cells was prepared following the methods to prepare cells. The cells were mixed with 200 pL of complete media and 50 pL of the cargo- loaded lipoCaP beads. The samples were placed into an incubator and were occasionally removed to take images over the course of 24 hours. The establishment of the networking system was observed after 30 minutes and cargo molecules were inside the cells at 3 hours.

Example 9. Cargo transfer from cells to beads

[00304] Cells were allowed to become around 60% confluent in a single well of a 6-well plate containing a volume of 2 mL of DMEM buffer. Then the cells were transfected with a solution containing 6 pL of the desired cargo molecule mixed with 3 pL of lipofectamine (Lipofectamine 3000, Thermo Fischer Scientific). After transfection, the cells were incubated for at least 12 hours, or until the cargo molecules was observed to be inside the cells. Then the cells were mixed with the lipoCaP beads and incubated for 3 hours. Transfer of the cargo from cells into the beads was observed after 3 hours

(FIG. 19)

Example 10. Animal models and tissue analysis

[00305] Skin tissue'. C57/B16 mice were injected subcutaneously with 50 pl of router solution. The animals were sacrificed 24 hrs after injection, the skin tissues were isolated and processed using standard protocols for embedding and cryosectioning. Slides were then imaged for fluorescence on a Zeiss Axiovert 200 Fluorescence microscope for the router membrane fluorophore and labeled mRNA cargo, (FIG. 26). DAPI staining was performed to identify nuclei.

[00306] Eye tissue'. C57/B16 mice were injected subconjunctivaly with 5 pl of router solution. The animals were sacrificed 24 hrs after injection and the eye tissues were isolated and processed using standard protocols for embedding and cryosectioning. Slides were then imaged for fluorescence on a Zeiss Axiovert 200 Fluorescence microscope for the router membrane fluorophore and labeled mRNA cargo (FIG. 27). DAPI staining was performed to identify nuclei.

[00307] Liver tissue: C57/B16 mice were injected locoregionally with 50 pl of router solution. The animals were sacrificed 24 hrs after injection and the liver tissues were isolated and processed using standard protocols for embedding and cryosectioning. Slides were then imaged for fluorescence on a Zeiss Axiovert 200 Fluorescence microscope for the router membrane fluorophore and labeled mRNA cargo (FIG. 28) and DAPI staining was performed to identify nuclei.

[00308] Pancreatic tissue: C57/B16 mice were injected locoregionally with 50 pl of router solution.

The animals were sacrificed 24 hrs after injection, the pancreas tissues were isolated and processed using standard protocols for embedding and cryosectioning. Slides were then imaged for fluorescence on a Zeiss Axiovert 200 Fluorescence microscope for the router membrane fluorophore and labeled mRNA cargo (FIG. 29). DAPI staining was performed to identify nuclei.

[00309] It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims.

Claims

CLAIMS WHAT IS CLAIMED IS:

1. An intercellular router construct system comprising:

(i) a cell;

(ii) an intercellular router construct which itself remains extracellular, comprising a containment layer, a networking element, and an internal infrastructure, and wherein the containment layer comprises lipids; and

(iii) a network that connects the cell and the intercellular router construct.

2. The intercellular router construct system of claim 1, wherein and the intercellular router construct optionally contains one or more cargo molecules.

3. An intercellular router construct system comprising:

(i) an intercellular router construct, comprising a containment layer, an internal infrastructure, and a networking element, wherein the internal infrastructure is located within the containment layer

(ii) a network connecting (a) a cell and the intercellular router construct, or (b) a plurality of intercellular router constructs; and

(iii) optionally, a cargo molecule.

4. An intercellular router construct system comprising:

(i) a cargo molecule;

(ii) an intercellular router construct comprising an internal infrastructure, a networking element, and a containment layer;

(iii) a network to and/or between a cell and/or other intercellular router constructs; and

(iv) an exchange of the cargo molecule.

5. The intercellular router construct system of any one of claims 1 to 4, wherein the containment layer is an extracellular containment layer.

6. The intercellular router construct system of any one of claims 1 to 5, wherein the containment layer insulates the internal infrastructure from a environment and prevents the leakage, loss, or exposure of the cargo into the extracellular environment.

7. The intercellular router construct system of any one of claims 2 to 6, wherein the containment layer comprises lipids.

8. The intercellular router construct system of claim 7, wherein the lipids are present in a lipid membrane.

9. The intercellular router construct system of claim 8, wherein the external surface of the intercellular router construct is covered by the membrane.

10. The intercellular router construct system of any one of claims 7 to 9, wherein the lipids comprise a Zwitterionic head group, a cationic head group, or an anionic head group, or any combinations thereof.

11. The intercellular router construct system of any one of claims 7 to 10, wherein the lipids comprise Zwitterionic lipids.

12. The intercellular router construct system of any one of claims 7 to 11, wherein the lipids comprise Zwitterionic lipids and membrane dopant lipids, wherein the total lipid composition of the containment comprises at most 5 mol%, 10 mol%, 15 mol%, 20 mol%, or 25 mol% of the membrane dopant lipids.

13. The intercellular router construct system of claim 7 to 10, wherein the lipids comprise cationic lipids.

14. The intercellular router construct system of any one of claims 7 to 10 or 13, wherein the lipids comprise cationic lipids and membrane dopant lipids, wherein the total lipid composition of the containment comprises at most 5 mol %, 10 mol%, 15 mol%, 20 mol %, or 25 mol% of the membrane dopant lipids.

15. The intercellular router construct system of claim 7 to 10, wherein the lipids comprise anionic lipids.

16. The intercellular router construct system of any one of claims 7 to 10 or 15, wherein the lipids comprise anionic lipids and dopant lipids, wherein the total composition of the containment comprises at most 5 mol%, 10 mol%, 15 mol%, 20 mol%, or 25 mol% of the dopant lipids.

17. The intercellular router construct system of any one of claims 7 to 16, wherein the lipids comprise Zwitterionic lipids, cationic lipids, or anionic lipids, or any combinations thereof, provided that the Zwitterionic lipids, cationic lipids, or anionic lipids are greater than or equal to 10 mol% of the total lipid content of the containment.

18. The intercellular router construct system of any one of claims 7 to 10, 13, 14, or 17, wherein the lipids comprise a single positive charge, a double positive charge, a triple positive charge, or a poly cationic charge.

19. The intercellular router construct system of any one of claims 7 to 10, 15, 16, or 17, wherein the lipids comprise a single negative charge, a double negative charge, a triple negative charge, or a polyanionic charge.

20. The intercellular router construct system of any one of claims 7 to 19, wherein the lipids comprise various head group sizes.

21. The intercellular router construct system of claim 20, wherein the head group sizes are about 40 to about 80 square Angstroms (A²).

22. The intercellular router construct system of any one of claims 7 to 21, wherein the lipids vary in shape.

23. The intercellular router construct system of claim 22, wherein the shape of the lipids comprise a cylindrical shape, a cone-shape, or an inverted cone-shape, or any combinations thereof.

24. The intercellular router construct system of claim 7 to 23, wherein the lipids comprise a phospholipid head group, a glycolipid head group, or a sterile head group, or any combinations thereof.

25. The intercellular router construct system of any one of claims 7 to 24, wherein the lipids comprise various chain lengths, number of tails, or number of double bonds, or any combinations thereof.

26. The intercellular router construct system of any one of claims 7 to 25, wherein the lipids comprise a polymerized lipid .

27. The intercellular router construct system of any one of claims7 to 26, wherein the lipids comprise a bilayer or a monolayer membrane, or any combination thereof.

28. The intercellular router construct system of claim 27, wherein the lipids are unilamellar or multilamellar.

29. The intercellular router construct system of any one of claims 7 to 28, wherein the lipids comprise soy phosphatidylcholine (Soy PC), dioleoyl phosphatidylcholine (DOPC), dioleoyl phosphatidylglycerol (DOPG), dioleoyl phosphatidylethanolamine (DOPE), dioleoyl trimethylammonium-propane (DOTAP), dioleyloxy-N-sperminecarboxamido ethyl-N,N-dimethyl- 1-propanaminium (DO SPA), dipalmitoylphosphatidylcholine (DPPC), hydrogenated soy phosphatidylcholine (HSPC), or heptatriacont tetraene dimethylamino butanoate (DLin-DMA), or any combination thereof.

30. The intercellular router construct system of any one of claims 1 to 29, wherein the containment comprises a polymer, a metallic substrate, a plastic, a ceramics, or a fatty acid, or any combinations thereof.

31. The intercellular router construct system of any one of claims 1 to 30, comprising a plurality of independent intercellular router constructs.

32. The intercellular router construct system of claim 31, wherein the containments of the plurality of intercellular router constructs are independently selected from a lipid, a polymer, a metallic substrate, a plastic, a ceramic, and a fatty acid.

33. The intercellular router construct system of claim 32, wherein the plurality of intercellular router constructs comprise a cationic lipid containment and an anionic lipid containment.

34. The intercellular router construct system of any one of claims 1 to 33, wherein the internal infrastructure comprises a geometry, a scale, a phase, a porosity, or a surface charge, or any combinations thereof.

35. The intercellular router construct system of claim 34, wherein the internal infrastructure comprises a spheroidal geometry, a planar geometry, a tubular geometry, or an irregular geometry.

36. The intercellular router construct system of claim 35, wherein the internal infrastructure is about 10 nm to 10 mm in largest dimension.

37. The intercellular router construct system of claim 36, wherein internal infrastructure is about 500 nm to 10 mm in largest dimension.

38. The intercellular router construct system of any one of claims 1 to 37, wherein the internal infrastructure comprises a solid body phase, a fluid body phase, or a semi-sold body phase.

39. The intercellular router construct system of claim 38, wherein the internal infrastructure comprises a solid body phase.

40. The intercellular router construct system of claim 39, wherein the solid body phase comprises an insoluble solid.

41. The intercellular router construct system of claim 40, wherein the insoluble solid is a salt.

42. The intercellular router construct system of claim 41, wherein the salt is a metal salt.

43. The intercellular router construct system of claim 42, wherein the metal salt comprises a divalent cation.

44. The intercellular router construct system of claim 43, wherein the metal salt comprises a calcium divalent cation or a magnesium divalent cation.

45. The intercellular router construct system of any one of claims 42 to 44, wherein the metal salt comprises a phosphate anion, a carbonate anion, or a sulfate anion, or any combinations thereof.

46. The intercellular router construct system of any one of claims 42 to 45, wherein the metal salt is calcium phosphate.

47. The intercellular router construct system of claim 40, wherein the insoluble solid is a metal.

48. The intercellular router construct system of claim 40, wherein the insoluble solid is a metal oxide.

49. The intercellular router construct system of claim 48, wherein the metal oxide is silica oxide.

50. The intercellular router construct system of claim 48, wherein the metal oxide is zinc oxide.

51. The intercellular router construct system of claim 40, wherein the insoluble solid comprises a solid polymer.

52. The intercellular router construct system of claim 39 wherein the solid body phase comprises an insoluble solid, a metal, a metal oxide, or a solid polymer, or any combinations thereof.

53. The intercellular router construct system of claim 38, wherein the infrastructure comprises a fluid body phase.

54. The intercellular router construct system of claim 53, wherein the fluid body phase comprises a physiological buffer.

55. The intercellular router construct system of claim 54, wherein the physiological buffer comprises a salt, a nutrient, a sugar, an amino acid, or a vitamin, or any combinations thereof.

56. The intercellular router construct system of claim 55, wherein the salt is NaCl.

57. The intercellular router construct system of claim 54 to 56, wherein the physiological buffer comprises a pH regulator.

58. The intercellular router construct system of claim 57, wherein the pH regulator comprises a phosphate salt.

59. The intercellular router construct system of claim 54 or 55, wherein the physiological buffer comprises a minimal essential media or an equivalent cell supporting basal media.

60. The intercellular router construct system of claim 53, wherein the fluid body phase comprises a nontoxic buffer.

61. The intercellular router construct system of claim 53 or 60, wherein the fluid body phase comprises a physiological buffer or a nontoxic buffer, or any combinations thereof.

62. The intercellular router construct system of claim 38, wherein the internal infrastructure comprises a semi-solid body phase.

63. The intercellular router construct system of claim 62, wherein the semi-solid body phase comprises a cross-linkage.

64. The intercellular router construct system of claim 63, wherein the cross-linkage comprises a physical cross-linkage or a chemical cross-linkage, or any combinations thereof.

65. The intercellular router construct system of any one of claims 32, 62, or 63, wherein the semi-solid body phase comprises a hydrogel.

66. The intercellular router construct system of claim 65, wherein the hydrogel comprises agarose, hyaluronans, chitosans, collagen, dextran, pectin, polylysine, polystyrene, gelatin, starch, polyvinylalcohol, poly(lactic-co-glycolic)acid (PLGA) polymers, (meth)acrylate-oligolactide-PEO- oligolactide-(meth)acrylate, polyethylene glycol) (PEO), polypropylene glycol) (PPO), PEO-PPO- PEO copolymers (Pluronic®), poly(phosphazene), poly(methacrylates), poly(N-vinylpyrrolidone), PL(G)A-PEO-PL(G)A copolymers, or poly(ethylene imine), or any combinations thereof.

67. The intercellular router construct system of any one of claims 38 to 66 wherein the internal infrastructure is porous.

68. The intercellular router construct system of any one of claims 38 to 67, wherein the internal infrastructure comprises an outer surface charge.

69. The intercellular router construct system of claim 68, wherein the surface charge comprises an ionic interaction or a polar covalent interaction, or any combinations thereof.

70. The intercellular router construct system of claim 69, wherein the surface charge comprises an ionic interaction.

71. The intercellular router construct system of claim 70, wherein the ionic interaction between the Zwitterionic lipids, the cationic lipids, or the anionic lipids, or any combinations thereof, and the surface of the internal infrastructure supports formation of the containment layer.

72. The intercellular router construct system of claim 69, wherein the surface charge comprises a polar covalent interaction.

73. The intercellular router construct system of claim 72, wherein the polar covalent interaction is between the Zwitterionic lipids, the cationic lipids, or the anionic lipids, or any combinations thereof, and the internal infrastructure of the containment further comprises an anchoring layer.

74. The intercellular router construct system of claim 73, wherein the anchoring layer is covalently bonded to the containment or the internal infrastructure, or any combinations thereof.

75. The intercellular router construct system of claim 74, wherein the anchoring layer comprises a hydrophobic anchoring element.

76. The intercellular router construct system of claim 74, wherein the hydrophobic anchoring element of the anchoring layer comprises a monomer, an oligomer, or a polymer, or any combinations thereof.

77. The intercellular router construct system of any one of claims74 to 76, wherein the anchoring layer comprises an alkyl chain or an alkenyl chain, or any combinations thereof.

78. The intercellular router construct system of any one of claims 74 to 77, wherein the anchoring layer comprises a Ci -is alkyl or a C2-18 alkenyl, or any combinations thereof, wherein each alkyl or alkenyl is optionally substituted with one or more substituents.

79. The intercellular router construct system of any one of claims 74 to 78, wherein the anchoring layer comprises hexylamine, octyl amine, decyl chloride, dodecyl amine, or octadecyl isocyanate, or any combinations thereof.

80. The intercellular router construct system of any one of claims 31 to 79, wherein the plurality of intercellular router constructs comprise different internal infrastructures. ,

81. The intercellular router construct system of claim 80, wherein the internal infrastructures comprise a hydrogel, calcium phosphate, calcium alginate, lithium iron phosphate, or SiCTC I S.

82. The intercellular router construct system of claim 80, wherein the internal infrastructures comprise a hydrogel or calcium phosphate.

83. The intercellular router construct system of any one of claims 2 to 82, wherein the networking comprises an array of connections to transfer the cargo molecule between the intercellular router construct and the cell, between the cell and the intercellular router, between intercellular router constructs, or between cells or any combinations thereof.

84. The intercellular router construct system of claim 2 to 83, wherein the networking element facilitates an interaction between the intercellular router construct and the cell.

85. The intercellular router construct system of any one of claims 2 to 84, wherein the networking element is more dense than water.

86. The intercellular router construct system of any one of claims 2 to 85, wherein the networking element and the intercellular router construct taken together are more dense than water.

87. The intercellular router construct system of any one of claims 2 to 86, wherein the networking element and the intercellular router construct further comprise a material that is more dense than water.

88. The intercellular router construct system of claim 87, wherein the material is miscible and the internal infrastructure of the intercellular router construct comprises a fluid body.

89. The intercellular router construct system of claim 87 or 88, wherein the material comprises a sugar, a protein, or a water soluble polymer, or any combinations thereof.

90. The intercellular router construct system of claim 89, wherein the sugar is sucrose.

91. The intercellular router construct system of claim 89 or 90, wherein the sugar is about 100 mM to 1 M.

92. The intercellular router construct system of claim 84, wherein the interaction between the intercellular router construct and the cell comprises an electrostatic affinity or a hydrophobic affinity.

93. The intercellular router construct system of claim 92, wherein the interaction between the intercellular router construct and the cell comprises an electrostatic affinity or a hydrophobic affinity between the containment layer and the cell.

94. The intercellular router construct system of any one of claims 12 to 93, wherein the dopant lipid comprises a cationic lipid.

95. The intercellular router construct system of claim 94, wherein the cationic lipid comprises dioleoyl phosphatidylethanolamine (DOPE), dioleoyl trimethylammonium-propane (DOTAP), or dioleyloxy-N-sperminecarboxamido ethyl-N,N-dimethyl-l-propanaminium (DOSPA), or any combination thereof.

96. The intercellular router construct system of any one of claims 12 to 93, wherein the dopant lipid comprises an anionic lipid.

97. The intercellular router construct system of claim 96, wherein the anionic lipid comprises dioleoyl phosphatidylglycerol (DOPG) or dioleoyl phosphatidylserine (DOPS), or any combinations thereof.

98. The intercellular router construct system of any one of claims 12 to 93, wherein the dopant lipid comprises a Zwitterionic lipid.

99. The intercellular router construct system of claim 98, wherein the Zwitterionic lipid comprises hydrogenated soy phosphatidylcholine (HSPC), soy phosphatidylcholine (Soy PC), or dioleoyl phosphatidylcholine (DOPC), or any combinations thereof.

100. The intercellular router construct system of any one of claims 12 to 93, wherein the dopant lipid comprises a Zwitterionic lipids, a cationic lipids, or an anionic lipids, or any combinations thereof.

101. The intercellular router construct system of any one of claims 1 to 100, wherein the intercellular router construct comprises a fusogenic element.

102. The intercellular router construct system of claim 101, wherein the fusogenic element comprises a dopant lipid, wherein the total lipid composition of the intercellular router comprises at most 5 mol%, 10 mol%, 15 mol%, 20 mol%, or 25 mol% of the dopant lipid.

103. The intercellular router construct system of claim 102, wherein the dopant lipid comprises a cationic lipid.

104. The intercellular router construct system of claim 103, wherein the cationic lipid comprisesdioleoyl phosphatidylethanolamine (DOPE), dioleoyl trimethylammonium-propane (DOTAP), or dioleyloxy-N-sperminecarboxamido ethyl-N,N-dimethyl-l-propanaminium (DOSPA), or any combination thereof.

105. The intercellular router construct system of claim 101, wherein the fusogenic element comprises a peptide.

106. The intercellular router construct system of claim 105, wherein the peptide is present at a concentration of 10 nM to 1 mM.

107. The intercellular router construct system of claim 105 or 106, wherein the peptide comprises a TAT cell-penetrating peptide or a KALA peptide, or any combination thereof.

108. The intercellular router construct system of claim 101, wherein the fusogenic element comprises a nanoparticle.

109. The intercellular router construct system of claim 108, wherein the nanoparticle is present at a concentration of 10 nM to 1 mM.

110. The intercellular router construct system of claim 108 or 109, wherein the nanoparticle comprises zinc oxide, silica oxide, or a lipoprotein particle, or any combination thereof.

111. The intercellular router construct system of claim 101, wherein the fusogenic element comprises a protein.

112. The intercellular router construct system of claim 111, wherein the protein is present at a concentration of 10 nM to 1 mM.

113. The intercellular router construct system of claim 111 or 112, wherein the protein comprises a SNARE coiled-coiled protein or a catechol protein, or any combinations thereof.

114. The intercellular router construct system of claim 101, wherein the fusogenic element comprises a lipid, a peptide, a nanoparticle, or a protein, or any combinations thereof.

115. The intercellular router construct system of any one of claims 1 to 114, wherein the intercellular router construct comprises a pore inducing element.

116. The intercellular router construct system of claim 115, wherein the pore inducing element comprises an exotoxin.

117. The intercellular router construct system of claim 116, wherein the exotoxin comprises a hemolysin.

118. The intercellular router construct system of claim 117, wherein the hemolysin is present at a concentration of 10 nM to 1 mM.

119. The intercellular router construct system of claim 117 or 118, wherein the hemolysin comprises an alpha hemolysin (SLO), or a cholesterol-dependent cytolysin, or any combination thereof.

120. The intercellular router construct system of any one of claims 1 to 119, wherein the intercellular router construct comprises a local environment modulator.

121. The intercellular router construct system of claim 120, wherein the local environment modulator comprises a local dehydration reagent.

122. The intercellular router construct system of claim 121, wherein the local dehydration reagent comprises PEG-3000 or a Ca²⁺ salt, or any combination thereof.

123. The intercellular router construct system of any one of claims 2 to 122, wherein the networking comprises an array of membrane bound tubules connecting an intercellular router construct with a cell.

124. The intercellular router construct system of claim 123, wherein the array of membrane bound tubules are on the order of 100 nm to 1 micron in width.

125. The intercellular router construct system of claim 123 or 124, wherein the array of membrane bound tubules are on the order of 100 nm to 200 microns in length.

126. The intercellular router construct system of any one of claims 2 to 124, wherein the cargo molecule comprises a small molecule, a large molecule, a protein, a lipid, a nucleic acid, a metabolite, an ion, a nutrient, or an organelle, or any combinations thereof.

127. The intercellular router of claim 126, wherein the cargo molecule comprises an exogenous material or a cell made material, or any combination thereof.

128. The intercellular router construct system of claim 126 or 127, wherein the cargo molecule comprises cytosolic material, nuclear material, other organelle localizing material, or membrane bound material, or any combination thereof.

129. The intercellular router construct system of claim 126, wherein the nucleic acid comprises DNA or RNA.

130. The intercellular router construct system of claim 129, wherein the RNA comprises mRNA, miRNA, anti-miRNA, or siRNA, or any combinations thereof.

131. The intercellular router construct system of claim 126, wherein the protein is a membrane receptor.

132. The intercellular router construct system of claim 131, wherein the membrane receptor is an immune antigen receptor.

133. The intercellular router of claim 132, wherein the immune antigen receptor is a chimeric antigen receptor.

134. The intercellular router construct system of any one of claims 1 to 133, wherein the intercellular router construct system comprises two or more cargo molecules.

135. The intercellular router construct system of any one of claims 1 to 134, wherein the intercellular router construct system further comprises a dispersing element.

136. The intercellular router construct system of claim 135, wherein the internal infrastructure fully enclosed by the containment layer is at least partially embedded within the dispersing element.

137. A method of establishing an intercellular router construct system comprising:

(i) contacting a cell with an intercellular router construct, wherein the intercellular network router construct comprises a containment layer comprising an internal infrastructure; thereby creating a network that connects the cell and the intercellular router construct and establishing an intercellular router construct system.

138. The method of claim 137, comprising extending a networking from the cell to the intercellular router construct, which connects the cell and the intercellular router construct.

139. The method of claim 138, further comprising extending a networking from the cell to a second cell, which connects the cell and the second cell.

140. The method of claim 137, comprising extending a plurality of networking from the cell.

141. The method of any one of claims 138 to 140, wherein the extending of the networking is stimulated by the presence of the intercellular router construct.

142. The method of any one of claims 137 to 141, wherein the cell is from the adipose, bone, breast, colon, esophagus, eye, heart, intestine, muscle, nasal cavity, kidney, liver, lung, oral cavity, ovary, pancreas, skin, or stomach.

143. The method of claim 142, wherein the cell is from the breast, colon, kidney, liver, pancreas, or skin.

144. The method of claim 142, wherein the cell is derived from the eye conjunctiva.

145. An intercellular router construct comprising: a) a containment layer; b) a localizing or dispersal element; and c) a composite infrastructure, wherein the composite infrastructure comprises at least a portion of the localizing or dispersal element and an internal infrastructure; wherein the containment layer encloses the composite infrastructure.

146. The intercellular router construct of claim 145, wherein the localizing or dispersal element and the internal infrastructure are in fluid communication.

147. The intercellular router construct of claim 145, wherein the localizing or dispersal element and the internal infrastructure are not in fluid communication.

148. The intercellular router construct of claim 145, wherein when a networking element is formed between the intercellular router and a cell, the networking element comprises two ends and one end connects to the internal infrastructure and one end connects to the cell.

149. The intercellular router construct of claim 148, wherein the networking element is formed through the localizing or dispersal element.

150. The intracellular router construct of claim 145, wherein the localizing element is a hydrogel.

151. The intracellular router construct of claim 145, wherein the dispersal element is a hydrocolloid.