+

WO2009052810A2 - Matrices de fibrilles de collagène - Google Patents

Matrices de fibrilles de collagène Download PDF

Info

Publication number
WO2009052810A2
WO2009052810A2 PCT/DE2008/001745 DE2008001745W WO2009052810A2 WO 2009052810 A2 WO2009052810 A2 WO 2009052810A2 DE 2008001745 W DE2008001745 W DE 2008001745W WO 2009052810 A2 WO2009052810 A2 WO 2009052810A2
Authority
WO
WIPO (PCT)
Prior art keywords
kollagenfibrillenmatrices
collagen
solution
varied
matrices
Prior art date
Application number
PCT/DE2008/001745
Other languages
German (de)
English (en)
Other versions
WO2009052810A3 (fr
Inventor
Babette Lanfer
Uwe Freudenberg
Ralf Zimmermann
Carsten Werner
Original Assignee
Leibniz-Institut Für Polymerforschung Dresden E.V.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Leibniz-Institut Für Polymerforschung Dresden E.V. filed Critical Leibniz-Institut Für Polymerforschung Dresden E.V.
Publication of WO2009052810A2 publication Critical patent/WO2009052810A2/fr
Publication of WO2009052810A3 publication Critical patent/WO2009052810A3/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/0068—General culture methods using substrates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14—Macromolecular materials
    • A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
    • A61L27/24—Collagen
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2533/00—Supports or coatings for cell culture, characterised by material
    • C12N2533/50—Proteins
    • C12N2533/54—Collagen; Gelatin

Definitions

  • the invention relates to collagen fibril matrices which are obtainable by a method for coating carriers with collagen fibril matrices of defined structure from flowing solutions, in which the size, shape and orientation of the resulting collagen fibrils and the coverage density of the carrier with collagen and the structure of the layers targeted by varying Parameters are controlled.
  • the invention is based in the field of biomaterials research, specifically the production of biological support materials as a scaffold for cell culture applications.
  • Collagen represents the quantitatively most abundant extracellular matrix fiber protein.
  • fibrillogenesis individual collagen molecules assemble into collagen fibrils. These collagen fibrils are usually heavily targeted in human and animal tissues. Examples include fiber bundles in muscle or tendon tissue.
  • the alignment of the fibrils fulfills special structural functions. For example, the load capacity of these tissues is significantly increased by the orientation.
  • the disadvantage of this method is, in particular, that the alignment of the collagen fibrils by use of the magnetic spheres results in significant changes in the properties of the collagen, whereby the resulting collagen gels are no longer in the native state. Thus, they are of limited use for cell experiments.
  • the aim of other methods is to directly generate collagen-oriented collagen structures, which systems can usually be defined as "collagen bands" in the form of a mononuclear layer with a layer thickness of about 3 nm over the substrate surface and a width of up to 20 nm et al., Journal of Structural Biology 148 (2004) 268-278 and Jiang et al., Microscopy Research and Technique 64 (2004) 435-440, alignment was accomplished in this approach by directionally pipetting collagen solution on a glow surface While the diameter of native collagen fibrils is 20 to 500 nm, the diameter range of the collagen bands thus obtained is only between 5 to 20 nm. On the other hand, for this method only surfaces of embers were used as surfaces.
  • the object underlying the invention is to provide collagen matrices for cell culture experiments in which cell-matrix interactions such as cell migration and adhesion can be investigated and a method for their production.
  • the method is intended to achieve the variation of the fibril shape, the density of the collagen fibrils on the substrate and the alignment of the fibrils, and in this way to adapt the collagen model surfaces to native conditions. Therefore, an essential part of the task is the targeted control of the morphology and orientation of the fibrils at the interface.
  • such model surfaces should be preparable on different carrier materials.
  • the object of the invention is to provide collagen fibril matrices obtainable from a process for coating carriers of collagen fibril matrices of defined structure from flowing solutions.
  • the group of parameters to be varied for control include:
  • the concentration of the solution the concentration of the solution
  • the collagen solution is conveyed by a pump through a microchannel, wherein the carrier to be coated forms the underside of the channel.
  • the fibrils of the collagen solution are aligned by the shearing action of the flowing liquid on the sample carrier.
  • the method according to the invention makes it possible to produce collagen fibril matrices that are controllable by setting defined parameters in their morphology.
  • the method is not limited only to the effect of aligning the collagen structures, but rather offers the possibility of producing different matrices in a collagen structure and by various parameters such as the shear conditions, the concentration / preparation of the solution and the substrate selection to control specifically.
  • the method according to the invention allows the targeted variation of the fibril form / morphology and thus offers the possibility of recreating native states by the generation of collagen model surfaces.
  • the methods mentioned in the prior art produce, inter alia, strongly artificial collagen matrices.
  • the "collagen band matrices" differ according to Jiang et al., Microscopy Research and Technique 64 (2004) 435-440, and those with magnetic spheres Guo and Kaufman's collagen matrices, Biomaterials 28 (2007) 1105-1114, are substantially different in structure and composition from native collagen fibril matrices.
  • Another advantage of the method according to the invention over the previously known methods is the significantly higher quality of the alignment. While the methods mentioned in the prior art achieve a coarse alignment of collagen gels, with the method according to the invention, individual fibrils can be specifically aligned on a preferably planar carrier.
  • the fibrillogenesis takes place in the flowing solution.
  • a cooled collagen solution is pumped from a reservoir through tempered tubes and heated at the same time. This leads to the beginning of the assembly process.
  • the collagen fibrils formed are then deposited by the defined overflow of the substrate surface in the microchannel and aligned at the same time.
  • the fibril size correlates with the fibrillation time and thus with the tube length and the flow rate in the system.
  • a prefibrated collagen solution is used.
  • the experimental setup is operated at constant temperature throughout the system. All other process parameters such as the shear rate and the dimensions of the channel are also constant. With this variant, matrices with very high orientation in the flow direction can be achieved.
  • the orientation of the collagen fibrils is controlled by the choice of flow rate.
  • the influencing of the occupation density is effected by the choice of the surface-active coating on the substrate.
  • the material for the surface-active coating is selected with regard to the hydrophobic or hydrophilic properties.
  • the substrate material used is glass.
  • a hydrophobic polymer thin film is used.
  • Poly (octadecene- ⁇ -maleic acid) (POMSA) is particularly suitable here.
  • the substrate material is provided with a surface-active coating in the form of a hydrophilic polymer thin film, preferably with poly (ethylene-aft-maleic acid) (PEMSA).
  • the resulting coverage density is influenced by the adjustment of the solution concentration.
  • solution concentrations for example, 0.2 mg / ml, 0.4 mg / ml or 0.8 mg / ml are used.
  • the collagen fibril matrices are applied by adsorption on preferably planar supports.
  • the occupation density is varied by the shear rate.
  • the orientation of the fibrils on the support can be varied while varying the fibril size by choosing different fibrillation times.
  • directed collagen fibrils are readjusted in vitro. These structures, as common extracellular matrix structures, are relevant to the study of cell-matrix interactions such as cell adhesion, proliferation, and differentiation.
  • FIG. 1 collagen matrices obtained from 0.8 mg / l of concentrated collagen solution at prefibrillation times of 5 min, 10 min and 30 min, FIG.
  • FIG. 2 collagen matrices obtained from 0.2 mg / ml, 0.4 mg / ml and 0.8 mg / ml concentrated collagen solution at a pre-fibrillation time of 5 minutes, FIG.
  • FIG. 3 collagen matrices obtained from prefibrillated collagen solution
  • FIG. 4 shows the influence of the substrate or of the surface-active coating on the occupation density on the basis of a diagram
  • Fig. 5 the alignment of collagen fibrils on various surface-active coatings.
  • the collagen fibril matrices according to the invention were obtained by two different process variants.
  • a cooled collagen solution is pumped out of the reservoir through tempered tubes and heated at the same time. This leads to the beginning of the assembly process.
  • the collagen fibrils formed are then deposited by the defined overflow of the substrate surface in the microchannel and aligned at the same time.
  • the fibril size correlates with the fibrillation time and thus with the tube length and the flow rate in the system. By increasing the collagen concentration, the density of collagen increases.
  • variant 2 an already fibrillated collagen solution is used.
  • the fibril solution is homogenized and centrifuged before use. This procedure ensures a uniform composition of the solution.
  • the experimental setup is operated at constant temperature throughout the system. All other process parameters such as the shear rate and the dimensions of the channel are also constant (see Table 1). With this variant, matrices with very high orientation in the flow direction can be achieved.
  • FIG. 1 shows images of collagen matrices produced with 0.8 mg / ml concentrated collagen solution according to Variant 1 initially cooled at 4 ° C. on a cleaned glass sample carrier with a hydrophobic polymer thin film poly (octadecene-aft-maleic acid) (POMSA) , The fibrils are formed during the flow in the tube, which was heated to 37 0 C.
  • the average residence time in the tube was two minutes in the experiments for the images in the first column (A) and 10 minutes in the experiments for the images in the second column (B); in the experiments to the figures of the third column (C) 30 min. This was followed by a one-hour flow through the canal.
  • FIG. 2 shows images of collagen matrices incubated with initially cooled at 4 ° C, (A) 0.2 mg / ml, (B) 0.4 mg / ml, (C) 0.8 mg / ml concentrated collagen solution
  • A 0.2 mg / ml
  • B 0.4 mg / ml
  • C 0.8 mg / ml concentrated collagen solution
  • POMSA hydrophobic polymer thin film poly
  • FIG. 3 contains images which show collagen matrices which were each produced with a collagen solution prefibrillated according to variant 2 on a cleaned glass sample carrier with a hydrophobic polymer thin film of poly (octadecene- ⁇ / f-maleic acid) (POMSA).
  • FIG. 4 shows a diagram showing the influence of the substrate or of the surface-active coating on the occupation density of collagen.
  • the following substrates were used in each case: cleaned glass sample carriers having a hydrophobic polymer thin film
  • FIG. 4 illustrates that as the hydrophobicity of the substrate surface increases, so does the resulting density of collagen.
  • Fig. 5 shows the respective orientation of collagen fibrils at the entrance, the center and the exit of the channel on the different surface active

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biochemistry (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Biophysics (AREA)
  • Dermatology (AREA)
  • Medicinal Chemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Transplantation (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Materials For Medical Uses (AREA)

Abstract

L'invention concerne des matrices de fibrilles de collagène qui peuvent être obtenues par un procédé de revêtement de supports avec des matrices de fibrilles de collagène de structure et de morphologie définies, à partir de solutions qui s'écoulent. La commande de la structure et de la morphologie s'effectue ici en faisant varier un ou plusieurs paramètres différents, sachant que le groupe des paramètres à faire varier comprend : - les matériaux de substrats; - les revêtements tensioactifs sur les matériaux de substrats; - la concentration de la solution; - le temps de fibrillation; et - les conditions marginales de dynamique des fluides du processus de séparation.
PCT/DE2008/001745 2007-10-24 2008-10-22 Matrices de fibrilles de collagène WO2009052810A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102007000564.6 2007-10-24
DE102007000564.6A DE102007000564B4 (de) 2007-10-24 2007-10-24 Verfahren zur Beschichtung von Trägern mit Kollagenfibrillenmatrices

Publications (2)

Publication Number Publication Date
WO2009052810A2 true WO2009052810A2 (fr) 2009-04-30
WO2009052810A3 WO2009052810A3 (fr) 2009-10-15

Family

ID=40467077

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DE2008/001745 WO2009052810A2 (fr) 2007-10-24 2008-10-22 Matrices de fibrilles de collagène

Country Status (2)

Country Link
DE (1) DE102007000564B4 (fr)
WO (1) WO2009052810A2 (fr)

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE327038T1 (de) * 2001-05-30 2006-06-15 Biolex Inc Testplatte für hochdurchsatzscreening
US20050019488A1 (en) * 2001-11-30 2005-01-27 Cambridge Polymer Group, Inc., Boston, Ma Layered aligned polymer structures and methods of making same
US7700333B2 (en) * 2004-07-26 2010-04-20 Agency For Science Technology & Research Immobilization of cells in a matrix formed by biocompatible charged polymers under laminar flow conditions

Also Published As

Publication number Publication date
DE102007000564A1 (de) 2009-04-30
WO2009052810A3 (fr) 2009-10-15
DE102007000564B4 (de) 2016-11-17

Similar Documents

Publication Publication Date Title
EP1457552B1 (fr) Support pour culture de cellules et tissus pour la préparation d'un implant et implant contenant cet support
EP2126166B1 (fr) Tissu non tissé à fibres disposées de manière aléatoire
DE69015284T2 (de) Poröse Membran mit einer Zellwachstum unterstützenden hydrophilen Oberfläche und Verfahren.
DE102006038703B4 (de) Verfahren und Vorrichtung zur Erzeugung von oxidischen Nanopartikeln aus einem Oxidpartikel bildenden Material
DE112016003589B4 (de) Künstliches blutgefäss, verfahren zur herstellung von künstlichem blutgefäss und verfahren zur herstellung eines poröses-gewebe-regenerationssubstrats
DE102010012845A1 (de) Durch Rotationsspinnverfahren hergestellte Mehrkomponentenfasern
DE102009013012A1 (de) Mit Therapeutika und Diagnostika beladene Kompositmaterialien umfassend Polymernanopartikel und Polymerfasern
EP1402055B1 (fr) Procede pour produire une couche de molecules fonctionnelles
DE102018129658B4 (de) Medizinisches Implantat umfassend Magnesium und Fibroin
DE102004047568A1 (de) Antimikrobielles Implantat mit einer flexiblen porösen Struktur
DE102007000564B4 (de) Verfahren zur Beschichtung von Trägern mit Kollagenfibrillenmatrices
EP3655575A1 (fr) Non-tissés avec imprégnation de silice asymétrique, procédé de fabrication desdits non-tissés et leur utilisation
Zhang et al. Effects of emulsion electrospinning parameters on the morphology and structure of core-shell structured PLLA fibers
EP4274928B1 (fr) Non-tissés de nanofibres à porosité élevée en tant que structure de support pour tissu stromale
DE102012111710B4 (de) Verfa zur Plasmabehandlung einer kolloidalen Lösung und Anwendung des Verfahrens
Boudriot et al. Elektrogesponnene Poly-L-Laktid-Nanofasern als resorbierbare Matrix für Tissue-Engineering/Electrospun Poly-l-lactide Nanofibres as Scaffolds for Tissue Engineering
EP2665810B1 (fr) Procédé pour l'immobilisation et la préparation de structures de multicomposants fonctionnelles de matrice extracellulaire
DE102006039588B4 (de) Substrat mit Lipidmembranen, Verfahren zur Erzeugung sowie Anordnung hierzu
DE102019123799B4 (de) Verfahren zur Herstellung von Biomaterialien mit porösen und glatten Topographien und deren Verwendung
DE102022134439B4 (de) Verfahren zur Herstellung eines faserverstärkten Hydrogelformkörpers
EP1177829B1 (fr) Membranes compatibles avec le sang et le tissu à base de copolymères d'acrylonitrile/N-vinylpyrrolidone et leur utilisation dans le domaine de la médecine
EP2628490A1 (fr) Matrice de colonisation cellulaire
WO2025051432A1 (fr) Biocomposite et son procédé de production
DE102006054789B4 (de) Verfahren und Vorrichtung zur Behandlung biologischer Zellen auf einem Substrat
DE202021106754U1 (de) Ein System zur Regeneration von Hautgewebe unter Verwendung von Nanomedizin und Nanofasern

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 08843164

Country of ref document: EP

Kind code of ref document: A2

122 Ep: pct application non-entry in european phase

Ref document number: 08843164

Country of ref document: EP

Kind code of ref document: A2

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载