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WO2007007452A1 - Vaisseau sanguin synthétique et procédé servant à produire celui-ci - Google Patents

Vaisseau sanguin synthétique et procédé servant à produire celui-ci Download PDF

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Publication number
WO2007007452A1
WO2007007452A1 PCT/JP2006/308293 JP2006308293W WO2007007452A1 WO 2007007452 A1 WO2007007452 A1 WO 2007007452A1 JP 2006308293 W JP2006308293 W JP 2006308293W WO 2007007452 A1 WO2007007452 A1 WO 2007007452A1
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WO
WIPO (PCT)
Prior art keywords
blood vessel
artificial blood
group
substrate
reactive functional
Prior art date
Application number
PCT/JP2006/308293
Other languages
English (en)
Japanese (ja)
Inventor
Tsutomu Furuzono
Kunio Miyatake
Ryoichi Tanaka
Masahiro Okada
Shoji Yasuda
Hiroyuki Kadono
Miwa Masuda
Original Assignee
Japan Health Sciences Foundation
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 Japan Health Sciences Foundation filed Critical Japan Health Sciences Foundation
Priority to JP2007524523A priority Critical patent/JP5028625B2/ja
Publication of WO2007007452A1 publication Critical patent/WO2007007452A1/fr

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Classifications

    • 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
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/507—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials for artificial blood vessels
    • 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
    • A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
    • A61L27/44—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
    • A61F2/02—Prostheses implantable into the body
    • A61F2/04—Hollow or tubular parts of organs, e.g. bladders, tracheae, bronchi or bile ducts
    • A61F2/06—Blood vessels

Definitions

  • the present invention relates to an artificial blood vessel capable of proliferating vascular endothelial cells to an artificial blood vessel at an early stage and having an ability closer to a real blood vessel, and a method for producing the same.
  • Patent Document 1 JP-A-5-344988 (published on December 27, 1993)
  • Patent Document 2 JP-A-8-332218 (published on December 17, 1996)
  • Non-Patent Document 1 Miho Tsuji et al., Artificial organ, 25, 208 (1996)
  • Non-Patent Document 2 Takahiro Tanikawa et al., Artificial Organ, 25, 224 (1996)
  • Non-Patent Document 3 N. Zempo, A.W.Clowes, Restenosis: pathogenesis and management,
  • Non-Patent Document 4 Permanent Oshima et al., Artificial organ, 23, 825 (1994)
  • Non-Patent Document 5 Kazue Katami et al., Artificial Organ, 25, 455 (1996)
  • Non-Patent Document 6 T.R.Kohler, et al., Surgery, 112, 901 (1992)
  • Non-Patent Document 7 Kazuyuki Ishibashi et al., Artificial Organ, 25, 733 (1966)
  • Non-Patent Document 8 Toshiya Shindo et al., Artificial organ, 25, 204 (1996)
  • Non-Patent Document 9 Y. Noishiki, et al., Nat. Med. 2, 90 (1996)
  • Non-Patent Document 4 Furthermore, in the artificial blood vessel whose surface is coated with gelatin, collagen, and albumin disclosed in Non-Patent Document 4, generation of unknown postoperative heat that seems to be a biological reaction to different types of gelatin and collagen has been observed. In other words, when biological materials are used, biological safety is a problem, which may cause infectious diseases.
  • Non-Patent Document 4 a human blood vessel whose surface is coated with gelatin, collagen, and albumin disclosed in Non-Patent Document 4 is also a large-diameter artificial blood vessel.
  • the bilayer-stretchable polytetrafluoroethylene human blood vessels disclosed in Non-Patent Documents 5 and 6 are not sufficient in antithrombogenicity and biological tissue compatibility, and in particular have an inner diameter of 6 mm.
  • the following vascular prosthesis does not provide a sufficient patency rate, and even in cases where transplantation is performed in humans, sufficient effects are not recognized, and future industrial use cannot be expected.
  • Non-Patent Documents 7 to 9 have problems such as difficulty in setting a cell collection source, complexity of a manufacturing method, difficulty in sterilization, and cost increase.
  • some artificial blood vessels such as polyester artificial blood vessels, require pre-clotting treatment to prevent leakage of artificial blood.
  • the pre-clotting process refers to the work of immersing the blood collected by the patient before implanting the artificial blood vessel. This puts a heavy burden on the patient and requires the development of an artificial blood vessel that does not require pre-crotting.
  • the present invention has been made in view of the above-mentioned problems, and the object of the present invention is to quickly proliferate vascular endothelial cells in an artificial blood vessel without using a material derived from an organism including animals. It is to provide an artificial blood vessel having high biocompatibility that can be obtained.
  • the method for producing an artificial blood vessel according to the present invention comprises at least one material selected from the group consisting of polyester-based resin, polyurethane-based resin and fluorine-based resin.
  • An artificial blood vessel manufacturing method in which an artificial blood vessel base material and biocompatible ceramic particles having biocompatibility are chemically bonded to each other, wherein the artificial blood vessel base material is hydrolyzed and treated.
  • the artificial blood vessel base material has a hydroxyl group generating step for generating a hydroxyl group on the surface of the material, a binder having a reactive functional group capable of being chemically bonded to the hydroxyl group and a reactive functional group capable of being chemically bonded to the biocompatible ceramic particles.
  • a reactive functional group introduction step of introducing a reactive functional group into the artificial blood vessel substrate by reacting with a hydroxyl group; and the reactive functional group and the biocompatible ceramic It is characterized by including a direct reaction step of reacting the particles, Ru.
  • the biocompatible ceramic is a (excellent) ceramic having high biocompatibility (histocompatibility, non-toxicity).
  • Specific examples of the biocompatible ceramics include at least one of calcium phosphate and titanium oxide.
  • the artificial blood vessel base material includes at least one material selected from the group consisting of a polyester-based resin, a polyurethane-based resin, and a fluorine-based resin.
  • This artificial blood vessel base material has a cylindrical shape, and includes, for example, a conventional artificial blood vessel made of polyester or polyurethane.
  • the artificial blood vessel substrate containing the material is subjected to hydrolysis treatment, Hydroxyl groups are generated on the surface of the artificial blood vessel substrate.
  • Hydroxyl groups are generated on the surface of the artificial blood vessel substrate.
  • the polymer chain of the material constituting the artificial blood vessel substrate is cut halfway, and a hydroxyl group is generated at the end of the cut polymer chain.
  • a reactive functional group capable of chemically bonding to the biocompatible ceramic particles is introduced into the artificial blood vessel substrate, and this reactive functional group and the biocompatible ceramic status are introduced. The particles are reacting.
  • an artificial blood vessel in which the biocompatible ceramic particles are chemically bonded to the surface of the artificial blood vessel substrate can be manufactured.
  • a hydroxyl group can be generated on the surface of the artificial blood vessel substrate. Therefore, the biocompatible ceramics can be produced using the hydroxyl group as a scaffold. Can be combined.
  • vascular endothelial cells When an artificial blood vessel is implanted in a living body (in a blood vessel), vascular endothelial cells can proliferate using the biocompatible ceramic particles as a scaffold. Due to the proliferation of the vascular endothelial cells, a thin, structurally stable intima is formed in the artificial blood vessel from the initial stage of implantation. Therefore, it is possible to obtain an artificial blood vessel that does not require a pre-crotting process.
  • biocompatible ceramic particles are chemically bonded to the surface of the artificial blood vessel, the biocompatible ceramic particles are rarely detached from the artificial blood vessel substrate force. For this reason, the proliferated vascular endothelial cells are rarely detached together with the biocompatible ceramics.
  • the vascular endothelial cells and the intercellular matrix exert an effect of entering the interface of the artificial blood vessel substrate (anchoring effect). For this reason, when the exfoliation of the thrombus in the artificial blood vessel is suppressed, the effect is exerted.
  • the human blood vessel according to the present invention can be suitably used not only for the above-mentioned large diameter (see Non-Patent Document 4) but also for an artificial blood vessel having a smaller inner diameter and a smaller diameter.
  • artificial blood vessels with an inner diameter exceeding S6mm are called “large-diameter artificial blood vessels”, those with a diameter exceeding 4mm and 6mm or less are called “medium-sized artificial blood vessels”, and those with a diameter of 4mm or less are called " It is called a small-diameter artificial blood vessel.
  • artificial blood vessels with a diameter of less than mm are sometimes referred to as “small-diameter artificial blood vessels”.
  • an artificial blood vessel by the above production method, it is possible to proliferate vascular endothelial cells early in the artificial blood vessel without using a living body-derived material.
  • An artificial blood vessel can be manufactured.
  • the method for producing an artificial blood vessel according to the present invention comprises at least one material selected from the group consisting of polyester-based resin, polyurethane-based resin, and fluorine-based resin.
  • a method for producing an artificial blood vessel in which an artificial blood vessel substrate and biocompatible ceramic particles having biocompatibility are chemically bonded to each other, wherein the artificial blood vessel substrate is hydrolyzed, and the artificial blood vessel substrate It comprises a hydroxyl group generating step for generating a hydroxyl group on the surface of the material, and a reaction step for reacting the hydroxyl group with a reactive group of a biocompatible ceramic particle having a reactive group capable of chemically bonding to the hydroxyl group.
  • the biocompatible ceramic particles having the reactive group on the surface of the artificial blood vessel can be obtained by hydrolyzing the artificial blood vessel substrate including the material.
  • a hydroxyl group for bonding can be generated.
  • an artificial blood vessel base material including at least one material selected from the group power consisting of polyester-based resin, polyurethane-based resin, and fluorine-based resin.
  • the polymer chain of the above material is cleaved in the middle to generate a hydroxyl group at the end of the cleaved polymer chain.
  • the biocompatible ceramic particle can be chemically bonded to the surface of the artificial blood vessel.
  • the biocompatible ceramic particles can be chemically bonded to the surface of the artificial blood vessel. Therefore, when the artificial blood vessel is implanted in the living body, the blood vessel endothelial cells are The biocompatible ceramic particles can be propagated as a scaffold.
  • the method for producing an artificial blood vessel according to the present invention includes at least one material selected from the group consisting of a polyester-based resin, a polyurethane-based resin, and a fluorine-based resin.
  • a binder functional group generating step for generating a reactive group capable of chemically bonding to the binder reactive functional group and the biocompatible ceramic particles A reactive functional group that introduces a reactive functional group into the artificial blood vessel substrate by reacting a binder having a reactive functional group capable of chemically bonding with the binder with the binder-reactive functional group of the artificial blood vessel substrate. It includes a group introduction step and a direct reaction step in which the reactive functional group reacts with the biocompatible ceramic particles.
  • a group force consisting of a butyl acetate and a (meth) acrylate ester group having a hydroxyl group is selected. At least one selected from By graft polymerization of the compound, a binder-reactive functional group containing a hydroxyl group that forms a chemical bond with the binder on the surface of the artificial blood vessel substrate is generated.
  • a binder-reactive functional group can be generated on the surface of the artificial blood vessel substrate by subjecting the artificial blood vessel substrate to surface treatment and then graft polymerization of the compound.
  • the biocompatible ceramic can be chemically bonded using the binder-reactive functional group as a scaffold.
  • vascular endothelial cells can proliferate using the biocompatible ceramic particles as a scaffold. Since biocompatible ceramic particles are chemically bonded to the surface of the artificial blood vessel, the biocompatible ceramic particles are unlikely to be detached from the artificial blood vessel substrate force. For this reason, the proliferated vascular endothelial cells are rarely detached together with the biocompatible ceramics.
  • an artificial blood vessel by producing an artificial blood vessel by the above production method, it is possible to proliferate vascular endothelial cells in the artificial blood vessel quickly without using a material derived from a living body.
  • An artificial blood vessel can be manufactured.
  • vascular endothelial cells When an artificial blood vessel is implanted in a living body (in a blood vessel), vascular endothelial cells can proliferate using the biocompatible ceramic particles as a scaffold. Due to the proliferation of the vascular endothelial cells, a thin, structurally stable intima is formed in the artificial blood vessel from the initial stage of implantation. Therefore, there is no need for pre-crotting processing.
  • biocompatible ceramic particles are chemically bonded to the surface of the artificial blood vessel, the biocompatible ceramic particles are rarely detached from the artificial blood vessel substrate force. For this reason, the proliferated vascular endothelial cells are rarely detached together with the biocompatible ceramics.
  • the human blood vessel according to the present invention can be suitably used not only for the above-mentioned large diameter (see Non-Patent Document 4) but also for an artificial blood vessel having a smaller inner diameter and a smaller diameter.
  • biological materials such as collagen are not used, biological safety is ensured, and further, sterilization, transportation, and storage are easy.
  • an artificial blood vessel by the above production method, it is possible to proliferate vascular endothelial cells early in the artificial blood vessel without using a biological material, and has high biocompatibility.
  • An artificial blood vessel can be manufactured.
  • the method for producing an artificial blood vessel according to the present invention includes at least one material selected from the group consisting of polyester-based resin, polyurethane-based resin, and fluorine-based resin.
  • a method for producing an artificial blood vessel in which an artificial blood vessel base material and biocompatible ceramic particles having biocompatibility are chemically bonded wherein after the surface treatment is performed on the artificial blood vessel base material, vinyl acetate, And a group force of hydroxyl group-containing (meth) acrylic ester, a binder-reactive functional group that generates a binder-reactive functional group on the artificial blood vessel substrate surface by graft polymerization of at least one selected compound.
  • Reaction of biocompatible ceramic particles having a production step, the binder-reactive functional group, and a reactive group capable of chemically bonding to the binder-reactive functional group It is characterized by including a reaction step of reacting the group, Ru.
  • the surface treatment is performed on the artificial blood vessel substrate containing the material, and then the compound is graft-polymerized to form the reactive group on the surface of the artificial blood vessel.
  • a binder-reactive functional group for chemically bonding the biocompatible ceramic particles can be generated.
  • the compound is graft-polymerized.
  • a binder-reactive functional group is generated on the surface of the human blood vessel substrate.
  • the biocompatible ceramic particles can be chemically bonded to the surface of the artificial blood vessel by reacting the binder monoreactive functional group with the reactive group of the biocompatible ceramic particles.
  • the biocompatible ceramic particles can be chemically bonded to the surface of the artificial blood vessel. Therefore, when the artificial blood vessel is implanted in the living body, the blood vessel endothelial cells are The biocompatible ceramic particles can be propagated as a scaffold.
  • vascular endothelial cells are placed in an artificial blood vessel.
  • the surface treatment is at least one surface selected from the group consisting of corona discharge treatment, ozone treatment, ultraviolet ray ozone treatment, and plasma treatment. Even processing.
  • an artificial blood vessel can be obtained that has an improved coverage rate of the biocompatible ceramic particles on the surface of the artificial blood vessel substrate.
  • the compound when corona discharge treatment is used, the compound can be graft polymerized by radical modification of the artificial blood vessel substrate surface. Then, the said compound is introduce
  • the plasma treatment is a surface treatment method using plasma, which is a gas containing charged particles separated from positive ions and electrons that travel around the nucleus due to discharge or the like and are separated from the atoms. .
  • the above plasma treatment is applied to nuclear fusion research and semiconductor manufacturing processes. Because of its low temperature controllability, the thin film by sputtering, etching or chemical vapor deposition (CVD) is used. Used for manufacturing. Therefore, the compound is introduced by performing a plasma treatment on the surface of the artificial blood vessel substrate to generate a binder-reactive functional group on the surface of the artificial blood vessel substrate.
  • Ozone has long been known as an oxidizing agent with a very high reaction activity, and ozone treatment using its acidity is "cleaning", “water purification”, “sterilization 'deodorization” Have already been widely applied to industries such as “bleaching”.
  • the compound is introduced onto the surface of the artificial blood vessel substrate, and a binder-reactive functional group is generated on the surface of the artificial blood vessel substrate.
  • Ultraviolet ozone treatment can exhibit a higher acidity than the ozone treatment by irradiating ozone gas with ultraviolet rays. Therefore, it has a binder-reactive functional group by performing ultraviolet ray ozone treatment on the surface of the artificial blood vessel substrate using ultraviolet ray-ozone treatment.
  • (Meth) acrylic acid ester or the like is introduced to generate a binder-reactive functional group on the surface of the artificial blood vessel substrate.
  • the (meth) acrylic acid ester having a hydroxyl group described above is butyl methacrylate 2-hydroxyethyl (hereinafter referred to as HEMA), acrylic acid 2- It may be at least one (meth) acrylic acid ester selected from the group consisting of hydroxymethyl and 2-hydroxypropyl methacrylate! /.
  • HEMA butyl methacrylate 2-hydroxyethyl
  • these (meth) acrylic acid esters are more preferably graft-polymerized with the artificial blood vessel base material. Therefore, the number of binder reactive functional groups generated on the surface of the artificial blood vessel substrate increases.
  • an artificial blood vessel can be obtained that has an improved coverage rate than the biocompatible ceramic particles on the surface of the artificial blood vessel substrate.
  • the reaction group introduction step of introducing the reaction group into the biofriendly ceramic particles may be performed before the reaction step.
  • the reactive group is introduced into the biocompatible ceramic particle, even the biocompatible ceramic particle having no reactive group may be the artificial blood vessel group. Can be chemically bonded to the material.
  • the artificial blood vessel substrate in the hydroxyl group generation step by the hydrolysis treatment, may be immersed in a basic solution within a range of pH 9 to pH 14. Good.
  • a hydroxyl group can be generated on the surface of the artificial blood vessel substrate by immersing the artificial blood vessel substrate in a basic solution in the range of pH 9 to pH 14.
  • the artificial blood vessel substrate can be hydrolyzed by immersing the artificial blood vessel substrate in the basic solution.
  • the hydroxyl group can be generated without impairing the physical properties of the artificial blood vessel substrate. it can.
  • hydroxyl group production by the hydrolysis treatment is performed.
  • a configuration in which the temperature of the basic solution is in the range of 10 ° C to 90 ° C is more preferable.
  • the artificial blood vessel substrate is hydrolyzed using the basic solution in the temperature range of 10 ° C to 90 ° C.
  • the hydroxyl group can be generated without further impairing the physical properties of the artificial blood vessel substrate by using the basic solution within the above temperature range.
  • the biocompatible ceramic particle is a calcium phosphate sintered body.
  • the calcium phosphate sintered body is obtained by sintering amorphous calcium phosphate at a high temperature and having high crystallinity. Since the calcium phosphate sintered body has high crystallinity, it can be left in the living body for a long period of time without dissolving even when implanted in the living body.
  • a calcium phosphate sintered body is used as the biocompatible ceramic particles.
  • the biocompatible ceramic particles are at least one of calcium phosphate and acid titanium, and the reactive functional group is an isocyanate group, an alkoxysilyl group, or a carboxyl group. And a structure that is at least one functional group selected from the group consisting of 4-methacryloxyltrimer trimethylate anhydride group.
  • At least one functional group selected from the group consisting of the isocyanate group, the alkoxysilyl group, the carboxyl group, and the 4-methacryloxy shetiltyl mellite antidelide basic group is the calcium phosphate and Z or acid titanium.
  • the calcium phosphate and Z or titanium oxide are used as the biocompatible ceramic particles, and the calcium phosphate and Z or titanium oxide is pretreated by introducing the functional group into the artificial blood vessel base material. Join without be able to.
  • the biocompatible ceramic particles have a particle size of ⁇ ! ⁇ 1 Used in the range of OOOnm.
  • the biocompatible ceramic particles and the artificial blood vessel substrate can be chemically bonded more firmly.
  • the biocompatibility ceramic particles are chemically bonded, so that characteristics such as elasticity of the artificial blood vessel base material are not impaired.
  • the artificial blood vessel base material is more preferably configured to have fiber strength.
  • the artificial blood vessel base material has a fiber strength.
  • a cylindrical artificial blood vessel substrate is constituted by a plurality of fibers.
  • an artificial blood vessel substrate having characteristics necessary for an artificial blood vessel substrate such as elasticity and rigidity can be used.
  • the artificial blood vessel according to the present invention is an artificial blood vessel in which biocompatible ceramic particles having biocompatibility are chemically bonded to the surface of an artificial blood vessel base material.
  • the lower limit of the coverage of the biocompatible ceramic particles on the vascular substrate surface is more preferably 10% or more, more preferably 20% or more, and particularly preferably 25% or more. When the coverage is 10% or less, the vascular endothelial cells do not sufficiently adhere to the artificial blood vessel surface.
  • biocompatible ceramic particles for example, nodyl apatite (HAp)
  • HAp nodyl apatite
  • the coverage of the biocompatible ceramic particles on the surface of the artificial blood vessel substrate is 10 Within the range of ⁇ 100%.
  • an artificial blood vessel substrate containing at least one material of a polyester-based resin and a polyurethane-based resin and biocompatible ceramic particles having biocompatibility are provided.
  • a method for producing an artificial blood vessel formed by chemical bonding comprising a step of hydrolyzing an artificial blood vessel substrate to generate a hydroxyl group on the surface of the artificial blood vessel substrate, and a chemical bond with the hydroxyl group.
  • the structure includes a reactive functional group introduction step for introducing an functional group and a direct reaction step for reacting the reactive functional group with the biocompatible ceramic particles.
  • an artificial blood vessel substrate containing at least one material of polyester-based resin and polyurethane-based resin and biocompatible ceramic particles having biocompatibility are provided.
  • a method for producing an artificial blood vessel formed by chemical bonding wherein the artificial blood vessel substrate is hydrolyzed to generate a hydroxyl group on the surface of the artificial blood vessel substrate, the hydroxyl group, And a reaction step of reacting the reactive group of the biocompatible ceramic particle having a reactive group capable of chemically bonding with the hydroxyl group.
  • an artificial blood vessel in which the biocompatible ceramic particles are chemically bonded to the surface of the artificial blood vessel substrate can be manufactured.
  • FIG. 1 is a drawing showing a scanning electron microscope image of an artificial blood vessel substrate in Example 1.
  • FIG. 2 is a drawing showing a fluorescence microscopic image showing the result of a cell adhesion test performed using the artificial blood vessel substrate of Example 1.
  • FIG. 3 is a drawing showing a fluorescence microscope image showing the results of a cell adhesion test conducted in Example 1 using a comparative artificial blood vessel substrate as a comparative example.
  • FIG. 4 is a drawing showing a fluorescence microscope image showing the results of a cell adhesion test conducted in Example 1 using a collagen-coated artificial blood vessel as a comparative example.
  • FIG. 5 is a drawing showing the results of confirming the introduction of hydroxyl groups by FT-IR ATR in Example 2.
  • FIG. 6 is a drawing showing a scanning electron microscope image of the surface of the HEMA artificial blood vessel substrate in Example 2.
  • FIG. 7 is a drawing showing the results confirmed by FT-IR ATR for introducing alkoxyl groups in Example 2.
  • FIG. 8 is a drawing showing a scanning electron microscope image of the KBE artificial blood vessel substrate in Example 2 in Example 2.
  • FIG. 9 is a drawing showing a scanning electron microscope image of an artificial blood vessel according to the present invention in Example 2.
  • FIG. 10 is a drawing showing the results of a cell adhesion test of an artificial blood vessel substrate according to a comparative example and the present invention in Example 2.
  • FIG. 11 is a drawing showing an appearance of an apatite-coated artificial blood vessel according to the present invention and a scanning electron microscope image of the surface of the artificial blood vessel in Example 3 in Example 3.
  • FIG. 12 is a drawing showing a state in which an apatite-coated human blood vessel and an untreated artificial blood vessel are implanted in a blood vessel of a dog's left carotid artery in Example 3.
  • Example 3 In Example 3, untreated artificial blood vessels and apatite-coated artificial blood vessels after 2 weeks from implantation were observed for appearance, and hematoxylin-eosin staining (hereinafter, “
  • FIG. 6 is a drawing showing the results of evaluation by “HE staining” and von Willebrand staining (hereinafter referred to as “VW staining”).
  • Example 3 the appearance of untreated artificial blood vessels and apatite-coated artificial blood vessels after 4 weeks of implantation, hematoxylin-eosin staining, and von Willebrand staining It is drawing which shows the result evaluated by (1).
  • FIG. 15 is a drawing showing the results of comparing the extension distances of vascular endothelial cells in an artificial blood vessel after implantation in Example 3.
  • FIG. 16 is a drawing showing the results of comparing the thickness of thrombus formed in an artificial blood vessel after implantation in Example 3.
  • FIG. 17 is a view showing the results of evaluation of an artificial blood vessel after 2 weeks from implantation by ⁇ -smooth muscle actin staining (hereinafter referred to as “oc-one SMA staining”) in Example 3.
  • FIG. 18 is a drawing showing the results of evaluation of an artificial blood vessel after a lapse of 4 weeks in Example 3 by a-SMA staining.
  • FIG. 19 is a drawing showing the appearance of Example 3 in which an apatite-coated artificial blood vessel and a collagen-coated artificial blood vessel have been cut after two weeks have passed since implantation.
  • the artificial blood vessel that works in the present embodiment has a configuration in which biocompatible ceramic particles having biocompatibility are chemically bonded to the surface of the artificial blood vessel base material. This will be explained below.
  • the artificial blood vessel substrate is one of the materials constituting the artificial blood vessel.
  • This artificial blood vessel base material is configured to include at least one material of polyester-based resin, polyurethane-based resin, and fluorine-based resin. More specifically, the artificial blood vessel substrate has the material on its surface. And the said artificial blood vessel base material has a cylindrical structure.
  • polyester resin examples include at least one of polyethylene terephthalate (PET), polybutylene terephthalate (PBT), polyester polyether block copolymer, and polyester-polyester block polymer. It is done.
  • polyurethane-based resin examples include at least one of polyurethane, a polyurethane material composed of diol and isocyanate, pandetas, diisocyanate polyether block copolymer, diisocyanate polyethylene glycol block copolymer, and the like.
  • Examples of the fluorinated resin include polytetrafluoroethylene (PTFE), tetrafluoroethylene / perfluoroalkyl butyl ether copolymer (PFA), and tetrafluoroethylene / hexafluoropropylene copolymer.
  • PTFE polytetrafluoroethylene
  • PFA perfluoroalkyl butyl ether copolymer
  • tetrafluoroethylene / hexafluoropropylene copolymer examples thereof include at least one of a polymer (FEP), tetrafluoroethylene / ethylene copolymer (ETFE), polyvinylidene fluoride (PVDF), polychloroethylene (PCTFE), and the like.
  • FEP polymer
  • ETFE tetrafluoroethylene / ethylene copolymer
  • PVDF polyvinylidene fluoride
  • PCTFE polychloroethylene
  • the material constituting the artificial blood vessel base material may be configured using only one of the polyester-based resin, the polyurethane-based resin, and the fluorine-based resin. Of these materials, a combination of multiple materials may be used.
  • the artificial blood vessel base material is more preferably constituted by using a plurality of fibers made of the above materials.
  • the plurality of fibers are processed by, for example, knitting, weaving, braiding, and more specifically, at least one of warp knitting, weft knitting, triaxial knitting, bag weaving, braiding, and the like. It is preferable that a cylindrical structure is formed by the method.
  • the structure of the artificial blood vessel base material is not limited to the above, and the tubular structure may be formed of, for example, a nonwoven fabric or the like.
  • an artificial blood vessel base material for example, a conventional artificial blood vessel made of polyester, polyurethane or PTFE can be used.
  • the materials constituting the artificial blood vessel substrate include materials other than the polyester-based resin, the polyurethane-based resin, and the fluorine-based resin (highly biocompatible material). It may be.
  • the material constituting the artificial blood vessel base material contains 50% by weight or more of the polyester-based resin, polyurethane-based resin and fluorine-based resin, and other materials are included. Also good.
  • the artificial blood vessel base material is constituted by a fiber cartridge, a fiber made of the polyester resin, a polyurethane resin and a fluorine resin and a fiber made of another material is used.
  • the artificial blood vessel base material may be formed.
  • the other material may be any material that constitutes a conventional artificial blood vessel that is generally available.
  • the artificial blood vessel substrate has a hydroxyl group on the surface thereof by performing a hydrolysis treatment. This hydrolysis treatment will be described later.
  • biocompatible ceramic particles will be described.
  • the above-mentioned biocompatible ceramics is a high biocompatibility (histocompatibility and / or nontoxicity) and (excellent) ceramic status.
  • Specific examples of the biocompatible ceramics include at least one of calcium phosphate and titanium oxide.
  • the titanium oxide is, for example, a compound represented by a chemical formula; TiO etc. It has a hydroxyl group on the surface of the compound.
  • the titanium oxide that is useful in the present embodiment refers to titanium oxide having a hydroxyl group on the surface.
  • titanium oxide is a compound represented by the chemical formula TiO
  • the titanium oxide titanium is suitable as a medical material because of its excellent affinity with living tissue, stability in the living environment and non-toxicity.
  • the calcium phosphate has high bioactivity, that is, biocompatibility with high biocompatibility.
  • Specific examples of the calcium phosphate include, but are not limited to, iodoxiapatite (Ca (PO) (OH)), tricalcium phosphate ( ⁇ (a) -tricalcium phosphate,
  • Ca (PO) calcium metaphosphate
  • Ca (OCP) calcium octaphosphate
  • the calcium phosphate is
  • calcium ion and a part of Z ion or phosphate ion are replaced by strontium ion, norlium ion, sodium ion, bicarbonate ion, carbonate ion, salt chloride ion, fluoride ion, etc. May be included.
  • a hydroxyapatite is suitable at a point with high biocompatibility among the said illustration.
  • the calcium phosphate it is more preferable to use a calcium phosphate sintered body (also called calcium phosphate ceramics) obtained by sintering calcium phosphate.
  • the calcium phosphate sintered body is less soluble in a living body having higher crystallinity than amorphous calcium phosphate.
  • the calcium phosphate sintered body can be obtained by sintering amorphous (amorphous) calcium phosphate. Specifically, for example, amorphous
  • the calcium phosphate sintered body can be obtained by sintering the calcium phosphate within a temperature range of 600 ° C. to 1300 ° C. for a predetermined time.
  • the crystallinity By sintering the calcium phosphate, the crystallinity can be increased, and for example, the solubility when introduced (implanted) into a living body can be reduced.
  • the degree of crystallinity of this calcium phosphate sintered body can be measured by X-ray diffraction (XRD). Specifically, the narrower the full width at half maximum of the peak indicating each crystal plane, the lower the crystallinity.
  • This Ca (PO) (OH) is a table of calcium phosphate.
  • the calcium phosphate may include a compound in which a hydroxide ion and / or a part of the phosphate ion of the calcium phosphate is substituted with a carbonate ion, a salt ion, a fluoride ion, or the like. Good. Further, the calcium phosphate may contain tricalcium phosphate and the like that are generated when amorphous hydroxyapatite is sintered.
  • the calcium phosphate sintered body is suitable as a medical material because of its excellent affinity with living tissue and stability in the living environment. Moreover, the calcium phosphate sintered body is difficult to dissolve in vivo. Therefore, the biological activity can be maintained for a long time in the living body.
  • the calcium phosphate sintered body according to the present embodiment can be obtained by sintering amorphous calcium phosphate.
  • the calcium phosphate may be artificially produced by a known production method such as a wet method, a dry method, a hydrolysis method, or a hydrothermal method. It may be.
  • the lower limit of the sintering temperature for sintering the amorphous calcium phosphate is more preferably 600 ° C or higher, and 700 ° C or higher is further preferable. Preferred is 800 ° C or higher. If the sintering temperature is lower than 600 ° C, sintering may not be sufficient. On the other hand, the upper limit of sintering temperature is 1300 ° C or less. More preferred is 1250 ° C or less, and even more preferred is 1200 ° C or less. If the sintering temperature is higher than 1300 ° C, calcium phosphate may decompose. Therefore, by setting the sintering temperature within the above range, it is possible to produce a calcium phosphate sintered body that is difficult to dissolve in the living body (high crystallinity).
  • the sintering time is not particularly limited and may be set as appropriate.
  • the dissolution rate of the calcium phosphate sintered body can be controlled. That is, by controlling the sintering temperature and the particle size, the physical properties of the calcium phosphate composite can be designed according to the application.
  • a hydroxyapatite sintered body or j8-tricalcium phosphate is used as a material constituting the calcium phosphate sintered body. It is more preferable.
  • no, idroxypatite sintered bodies are difficult to dissolve in vivo. Therefore, for example, when an artificial blood vessel is manufactured using the hydroxyapatite sintered body, the biological activity can be maintained for a long time in vivo.
  • the shape of the biocompatible ceramic particles is particulate.
  • the particle size of the particles may be a particle size that can be fixed to the surface of the artificial blood vessel substrate by chemically bonding the biocompatible ceramic particles and the artificial blood vessel substrate.
  • the lower limit of the particle diameter is more preferably 0.001 ⁇ m or more, and even more preferably 0.01 ⁇ m or more.
  • the upper limit of the particle size is preferably 1000 m or less, more preferably 100 m or less, and particularly preferably L m or less.
  • the term "particle size” refers to the shape of the particle.
  • the diameter of the largest inscribed circle is intended, for example, if the particle shape is substantially circular, the diameter of the circle is intended, and if it is substantially elliptical, the minor axis of the ellipse is Intended if the shape is substantially square, the length of the side of the square is intended; if the shape is substantially rectangular, the length of the short side of the rectangle is intended.
  • the “particle diameter” may be measured with a scanning electron microscope (SEM) or the like.
  • the “particle diameter” may be determined by measuring the particle diameter of arbitrarily selected particles and calculating the average value.
  • the binder has a reactive group capable of reacting with a hydroxyl group and a reactive functional group capable of binding to the biocompatible ceramic particles.
  • the reactive group is not particularly limited as long as it can react with a hydroxyl group. Specific examples thereof include an alkoxysilyl group, an isocyanate group, and a carboxyl group.
  • the reactive functional group is not particularly limited as long as it is capable of chemically bonding to the biocompatible ceramic particles.
  • the biocompatible ceramic particles may be an acid group.
  • the reactive functional group includes at least one of an isocyanate group, an alkoxysilyl group, a carboxyl group, and a 4-methacryloxetyl trimellrate anhydride group. There are two functional groups.
  • binder having the reactive group and the reactive functional group include a silane coupling agent having the reactive group.
  • silane coupling agent having this reactive group include ⁇ -methacryloxypropyltrimethoxysilane, y-methacryloxypropyltriethoxysilane, butyltrimethoxysilane, buturetriethoxysilane, p-styrylmethoxysilane, ⁇ -methacryloxy.
  • propylmethyldimethoxysilane, ⁇ -methacryloxypropylmethyljetoxysilane, ⁇ -ataryloxypropyltrimethoxysilane and the like can be used.
  • the binder is It has a alkoxysilyl group.
  • alkoxysilyl group refers to a group containing Si-OR (R represents an alkyl group).
  • SiH— (OR), —SiH— (OR) and the like are also included in the alkoxysilyl group. And above Si
  • R in OR represents an alkyl group or hydrogen.
  • the hydroxyloxysilyl group is also a kind of alkoxysilyl group.
  • the alkyl group a methyl group, an ethyl group, and the like can be preferably used.
  • the reactive functional group can be introduced into the artificial blood vessel substrate.
  • a hydroxyl group generating step is performed in which the artificial blood vessel substrate is hydrolyzed to generate a hydroxyl group on the surface of the artificial blood vessel substrate.
  • the reactive functional group is introduced onto the surface of the artificial blood vessel substrate by performing a reactive functional group introduction step of reacting the hydroxyl group present on the surface of the artificial blood vessel substrate with the binder. After the introduction of a group, a direct reaction step is performed in which the reactive functional group reacts with the biocompatible ceramic particle.
  • a reactive functional group that reacts the biocompatible ceramic particle with the binder is performed. After introducing the reactive group on the surface of the biocompatible ceramic particles by performing a process, the reaction process of reacting the reactive group with the hydroxyl group present on the surface of the artificial blood vessel substrate is performed. It is. In the above method (2), when the biocompatible ceramic particles have the reactive group as the initial force, the reactive group introduction step may not be performed.
  • Each step is explained below [0117] [Hydroxyl generation step by hydrolysis]
  • hydroxyl groups are generated on the surface of the artificial blood vessel substrate by hydrolyzing the artificial blood vessel substrate. More specifically, hydrolysis is performed on at least one polymer material in which the group power composed of the polyester-based resin, the polyurethane-based resin, and the fluorine-based resin constituting the artificial blood vessel base material is also selected. By doing this, the main chain of the polymer material is partially cut, and the cut end is a hydroxyl group.
  • the artificial blood vessel base material is formed by performing a hydrolysis treatment, and the polyester-based resin, the polyurethane-based resin, and the polymer chain of Z or fluorine-based resin. (Main chain) is cleaved in the middle, and the cleaved end is used as a hydroxyl group.
  • the main chain of the resin existing on the surface of the artificial blood vessel substrate is cut in the middle, and the cut end is used as a hydroxyl group.
  • hydroxyl groups are present on the surface of the artificial blood vessel substrate.
  • the artificial blood vessel having a hydroxyl group on the surface is controlled by controlling the hydrolysis conditions.
  • the physical properties of the substrate can be controlled. That is, by performing the above hydrolysis according to the hydrolysis conditions, the number of hydroxyl groups to be generated and physical properties such as elasticity and rigidity of the artificial blood vessel substrate after the hydrolysis treatment can be controlled. wear.
  • the hydrolysis conditions are described in detail below.
  • the artificial blood vessel substrate may be immersed in a basic solution (alkaline solution). More specifically, the artificial blood vessel substrate may be immersed in a basic solution having a predetermined concentration while stirring the artificial blood vessel substrate at a predetermined temperature for a predetermined time!
  • a basic solution alkaline solution
  • the artificial blood vessel substrate may be immersed in a basic solution having a predetermined concentration while stirring the artificial blood vessel substrate at a predetermined temperature for a predetermined time!
  • Examples of the solute constituting the basic solution include potassium hydroxide, sodium hydroxide, lithium hydroxide, calcium hydroxide, calcium hydroxide magnesium aqueous ammonia solution, carboxylic acid and Strong alkali salts such as dicarboxylic acid (sodium, potassium, etc.), oleic acid Examples include sodium RCOONa (R represents an alkyl group), strong alkali salts of higher fatty acids (sodium, potassium, etc.), sulfonic acid-based strong alkali salts (sodium, potassium, etc.), and aminic bases.
  • the solvent constituting the basic solution is not particularly limited, and may be appropriately selected depending on the kind of the solute and the material constituting the artificial blood vessel base material. Specifically, water, an organic solvent, etc. are mentioned.
  • the basic solvent is more preferably a sodium hydroxide aqueous solution which is preferably a basic aqueous solution because the material constituting the artificial blood vessel substrate is an organic substance.
  • the normality of the basic solution is more preferably in the range of 1 X 10 to 5 to 1N, and more preferably in the range of 1 X 10 to 4 to 0.5N 1 X 10 _3
  • the range of ⁇ 0.25N is particularly preferred.
  • the normality of the basic solution is greater than 1N, the physical properties of the artificial blood vessel substrate that has generated hydroxyl groups cannot be maintained, and if the final artificial blood vessel function is not exhibited, it is not a force but an artificial material. Molecules may fall off the vascular substrate.
  • the hydrolysis treatment cannot sufficiently generate hydroxyl groups on the surface of the artificial blood vessel substrate, and the artificial blood vessel group In some cases, the material and the biocompatible ceramic particles cannot be sufficiently chemically bonded.
  • the pH of the basic solution used for the hydrolysis treatment is preferably in the range of pH 9 to pH 14, more preferably in the range of pH 10 to pH 13.75, and ⁇ 11 to ⁇ 13. Within the range of 5 is particularly preferred.
  • the pH of the basic solution is higher than 14, it becomes difficult to make the physical properties of the artificial blood vessel substrate that has generated hydroxyl groups suitable for use as an artificial blood vessel, and the artificial manufactured finally produced. It may not function as a blood vessel. In addition, molecules may fall off the surface of the artificial blood vessel substrate.
  • the pH of the basic solution is smaller than 9, hydroxyl groups cannot be sufficiently generated on the surface of the artificial blood vessel substrate by the hydrolysis treatment, and the artificial blood vessel substrate and the biocompatible ceramics are not produced. In some cases, the particles cannot be sufficiently chemically bonded.
  • the temperature at which the water-hydrolysis is performed is more preferably in the range of 10 ° C to 90 ° C. More preferably within the range of ° C to 80 ° C, particularly preferably within the range of 30 to 70 ° C. Perform the above hydrolysis When the temperature is lower than 10 ° C, the hydrolysis of the artificial blood vessel substrate may not sufficiently proceed, and the desired number of hydroxyl groups generated on the surface of the artificial blood vessel substrate is generated. May not. In addition, when the hydrolysis temperature is 10 ° C.
  • the stirring conditions at this time are more preferably conditions that allow the basic solvent to convection around the artificial blood vessel substrate so that the artificial blood vessel substrate can be uniformly hydrolyzed. ,.
  • the solution may be stirred using a stirrer or the like.
  • the stirring speed (starter rotation speed) at this time is preferably 400 rpm or less, more preferably 300 rpm or less, and even more preferably 200 rpm or less. If the stirring speed is 400 rpm or more, the physical properties of the artificial blood vessel substrate may be impaired.
  • a binder-reactive functional group containing a hydroxyl group that forms a chemical bond with a binder can be generated on the surface of the artificial blood vessel substrate, the method for generating a hydroxyl group by hydrolysis described above can be used. After the surface treatment of the artificial blood vessel substrate, which is not limited, a method of graft polymerization of at least one compound selected from the group strength of vinyl acetate and (meth) acrylate having hydroxyl group is selected. Even if it is used, a binder-reactive functional group can be suitably generated for an artificial blood vessel substrate.
  • corona discharge treatment is used as a method for surface treatment of the artificial blood vessel substrate, A method of using HEMA as the (meth) acrylic acid ester containing a hydroxyl group will be described.
  • the coronal discharge treatment is performed on the artificial blood vessel substrate, and then graft polymerization with HEMA is performed to generate a binder-reactive functional group on the surface of the artificial blood vessel substrate. More specifically, by performing corona discharge treatment on at least one polymer material selected from the polyester-based and polyurethane-based resins constituting the artificial blood vessel substrate. After the surface of the material is radically modified, HEMA is graft polymerized to generate hydroxyl groups on the artificial blood vessel substrate surface.
  • HEMA is graft-polymerized to radically modify the surface of the artificial blood vessel substrate, and the HEMA having a hydroxyl group is graft-polymerized, whereby A hydroxyl group is generated on the surface of the blood vessel substrate.
  • hydroxyl groups exist on the surface of the artificial blood vessel base material.
  • the human blood vessel base material constitutes a polyester-based resin, a polyurethane-based resin, and / or a fluorine-based resin. It is possible to produce hydroxyl groups without degrading physical properties such as the tensile strength of fat.
  • the method of corona discharge treatment is not particularly limited, and an existing corona discharge treatment apparatus may be appropriately employed.
  • the execution value of alternating current in a standard gas (20 ° C, 1 atm) is preferably 20 KVZcm to 10000 KVZcm, more preferably 22 KVZcm to 500 OKV / cm, and particularly preferably 25 KVZcm to 1000 VZcm.
  • the electrode may have heat and the physical properties of the substrate may be impaired.
  • corona discharge does not start and surface modification of the equipment may not be possible.
  • the specific method for graft polymerization of HEMA and the corona discharge-treated artificial blood vessel substrate is not particularly limited, and may be carried out after considering appropriate optimum conditions.
  • the polymerization temperature is preferably 0 ° C to 200 ° C, more preferably 5 ° C to 150 ° C. Most preferably between 10 ° C and 100 ° C. If the temperature is 200 ° C or higher, the physical properties of the base material itself may be impaired. If the temperature is lower than o ° c, corona discharge is performed in the atmosphere. The surface treatment may not be possible.
  • the atmosphere for carrying out the grafting reaction may be air, an inert gas atmosphere such as nitrogen gas, or argon gas, but it is more preferably standard air to induce corona discharge.
  • the reaction time of the graft reaction is preferably a force of 1 minute to 3000 minutes, which can be adjusted according to the target hydroxyl group production amount, more preferably 5 minutes to 600 minutes, and particularly preferably 10 minutes to 120 minutes. It is. If it is longer than 3000 minutes, the graft polymerization may progress too much and the physical properties of the substrate may deteriorate, and if it is shorter than 1 minute, sufficient hydroxyl groups may not be introduced.
  • corona discharge treatment is used as a method for surface treatment of the artificial blood vessel substrate.
  • the method is not limited thereto.
  • plasma treatment, ozone treatment, or ultraviolet ozone treatment may be used.
  • HEMA was used as the compound to be graft polymerized, it is not limited to this.
  • a (meth) acrylic acid ester having a hydroxyl group such as 2-hydroxymethyl acrylate and 2-hydroxypropyl methacrylate is used. It is also possible to use butyl acetate.
  • the human blood vessel base material after the binder reactive functional group generation step is reacted with the binder having the reactive functional group, more specifically, the surface of the artificial blood vessel base material is present.
  • the reactive functional group is introduced into the surface of the artificial blood vessel base material by reacting the binder reactive functional group being reacted with the reactive group possessed by the binder.
  • the silane coupling agent has a chemical structure as shown in chemical formula (1).
  • Z include reactive groups capable of reacting with the hydroxyl groups of the artificial blood vessel substrate, and specific examples include (meth) ataryloxy groups, vinyl groups, and gen groups.
  • OR a methoxy group, an ethoxy group, etc. are mentioned, for example.
  • Z and Si which are reactive groups in the chemical formula (1), may be bonded by a high molecular chain or a low molecular chain, or may be directly bonded.
  • silane coupling agent specifically, for example, ⁇ -methacryloxypropyltrimethoxysilane, ⁇ -methacryloxypropyltriethoxysilane, butyltrimethoxysilane, vinyltriethoxysilane, ⁇ -styrylmethoxy
  • examples thereof include silane, ⁇ -methacryloxypropylmethyldimethoxysilane, ⁇ -methacryloxypropylmethyl jetoxysilane, y-ataryloxypropyltrimethoxysilane, and the like.
  • an alkoxysilyl group may be introduced into the artificial blood vessel substrate by polymerizing the silane coupling agent and the artificial blood vessel substrate in the presence of a polymerization initiator and a solvent. it can.
  • solvents such as benzyl alcohol, methanol and toluene are preferably used.
  • polymerization initiator for example, hydrogen peroxide or the like may be used.
  • the lower limit of the amount of silane coupling agent used is preferably 10% by weight or more, more preferably 50% by weight or more, more preferably 100% by weight with respect to the artificial blood vessel substrate. An amount of at least% is particularly preferred. If the amount used is less than 10% by weight, a sufficient amount of the alkoxysilyl group may not be introduced.
  • the upper limit of the amount used is preferably 500% by weight or less, more preferably 400% by weight or less, more preferably 300%. U, especially preferred below. The amount used is more than 500% by weight, and it is economical!
  • the polymerization is more preferably performed in a nitrogen atmosphere.
  • the lower limit of the polymerization temperature is more preferably 20 ° C or higher, more preferably 0 ° C or higher, and particularly preferably 20 ° C or higher.
  • the upper limit of the polymerization temperature is more preferably 120 ° C. or less, more preferably 100 ° C. or less, and still more preferably 90 ° C. or less. If the polymerization temperature is higher than 120 ° C, the polymerization amount cannot be controlled.
  • the polymerization time may be appropriately set so as to achieve a desired introduction rate (ratio at which alkoxysilyl groups are introduced into the artificial blood vessel substrate)!
  • the lower limit of the introduction ratio (% by weight) of the reactive functional group to the artificial blood vessel substrate is preferably 0.1% by weight or more, more preferably 1% by weight or more.
  • the introduction rate is the ratio of the weight of the silane coupling agent introduced per unit weight of the artificial blood vessel substrate.
  • the upper limit of the introduction rate is not particularly limited, but when the introduction rate is higher than 100% by weight, the amount of biocompatible ceramic particles that bind to the artificial blood vessel substrate becomes too large. It may not be economical.
  • the introduction rate of the reactive functional group may be appropriately set according to the purpose of use of the finally obtained artificial blood vessel.
  • a reactive group (not an alkoxysilyl group) possessed by the silane coupling agent for introducing an alkoxysilyl group into an artificial blood vessel base material and a hydroxyl group of the artificial blood vessel base material are reacted in an aqueous solvent.
  • the alkoxysilyl group of the silane coupling agent by protecting the alkoxysilyl group of the silane coupling agent with a surfactant, the alkoxysilyl group can be introduced into the artificial blood vessel substrate even in an aqueous solvent.
  • the reactive functional group is a sulfonate group.
  • an isocyanate group is introduced into an artificial blood vessel base material by polymerizing it with the above-mentioned binder having an isocyanate group and a reactive group, which are reactive functional groups, and an artificial blood vessel base material, the isocyanate group is contained in the reaction solvent. Reacts in active solvents such as dehydrated dimethyl sulfoxide, dehydrated dimethylformamide, etc. It is preferable to make it.
  • the isocyanate group is added to the artificial blood vessel substrate by reacting the reactive group of this binder with the hydroxyl group of the artificial blood vessel substrate and then heating. Can be introduced.
  • the blocking agent for example, when phenol is used, the blocking agent protecting the isocyanate group can be eliminated by heating within a range of 110 to 120 ° C.
  • the blocking agent for example, when imidazole is used, the blocking agent is heated within the range of 110 to 130 ° C, and when oxime is used, the heating is performed within the range of 130 to 150 ° C.
  • Specific examples of the blocking agent include phenol-containing compounds such as methyl salicylate and methyl-p-hydroxybenzoate; imidazole; oxime-containing compounds such as methyl ethyl ketoxime and acetone oxime, and the like. Can be mentioned.
  • N-hydroxyimide-containing compounds such as N-hydroxyphthalimide and N-hydroxysuccinimide
  • alcohols such as methoxypropanol, ethylhexanol, pentol, and ethyllatate Containing compounds
  • Ratatam-containing compounds such as force prolatatam and pyrrolidinone
  • active methylene compounds such as ethyl acetate and the like
  • the artificial blood vessel substrate into which the reactive functional group is introduced and the biological parent Directly reacts with compatible ceramic particles.
  • an artificial blood vessel in which the biocompatible ceramic particles and the artificial blood vessel substrate are bonded by chemical bonding can be manufactured.
  • a hydroxyapatite sintered body (hereinafter referred to as an "introduced product") having an alkoxysilyl group introduced therein with a hydroxyapatite sintered body, for example, the hydroxyapatite is introduced into the introduced product. What is necessary is just to heat, after making a sintered compact adsorb
  • the hydroxyapatite sintered body is adsorbed on the surface of the introduced material by immersing the introduced material in a dispersion in which the hydroxyapatite sintered body is dispersed. Then, the nodoxyapatite sintered body adsorbed on the surface is reacted with the alkoxysilyl group by heating.
  • dispersion medium for dispersing the above-mentioned nodyl and adroxyapatite sintered body include, for example, hydrocarbon solvents such as toluene and hexane; alcohols; ethers such as tetrahydrofuran and diethyl ether. Solvents; Organic solvents such as ketone solvents such as acetone and methyl ethyl ketone; Of the above-exemplified solvents, alcohols are preferably used in that the hydroxyapatite sintered body is satisfactorily dispersed and the surfactant protecting the alkoxysilyl group is eliminated.
  • dispersion media only one kind may be used, or a plurality of dispersion media may be used in combination.
  • a hydrocarbon solvent such as hexane or toluene
  • a stirrer such as a stirrer
  • Dispersion using an ultrasonic device (3) Combined use of the stirring device and the ultrasonic device, etc. may be used.
  • the lower limit of the additive amount of the sintered nodyloxyapatite is preferably 0.01% by weight or more with respect to the dispersion medium. More than 02% by weight is more preferable 0.05% by weight is particularly preferable. If the added amount of the hydroxyapatite sintered body is less than 0.01% by weight, the hydrated xanthite sintered body may not be uniformly adsorbed on the surface of the introduced product, and a uniform coated surface may not be formed. is there. on the other hand, As the upper limit of the amount of the hydroxyapatite sintered body added,
  • the amount added is more than 5.0% by weight, the amount of the hydroxyapatite sintered body remaining in the dispersion is remarkably larger than the amount of the hydroxyapatite sintered body adsorbed on the surface of the introduced product. Increased and not economical.
  • the lower limit of the reaction temperature at which the hydroxyl group of the hydroxyapatite sintered body adsorbed on the surface of the introduced product reacts with the alkoxysilyl group introduced into the introduced product is more preferably 25 ° C or more. More preferably, 50 ° C or higher is more preferable. 80 ° C or higher is particularly preferable. If the reaction temperature is lower than 25 ° C, the hydroxyapatite sintered body and the alkoxysilyl group may not react. On the other hand, the upper limit of the reaction temperature is particularly preferably 150 ° C or less, more preferably 200 ° C or less, more preferably 175 ° C or less. When the reaction temperature is higher than 200 ° C, the introduced product may decompose.
  • the introduced substance it is more preferable to wash the introduced substance with the same solvent as the dispersion medium after immersing the introduced substance in the dispersion and before reacting. Since the surface of the introduced substance after being immersed in the dispersion liquid is laminated with a sinter and idroxyapatite sintered body, and reacted without washing, the sinter and idroxyapatite sintered body are laminated. The physical properties of the introduced product may be impaired.
  • the reaction may be carried out under vacuum conditions.
  • a hydroxyapatite sintered body By reacting a hydroxyapatite sintered body with an alkoxysilyl group under vacuum conditions, a hydroxyapatite composite can be produced more quickly.
  • the pressure for performing the reaction is preferably in the range of 0. OlmmHg d. 33 kPa) to 10 mmHg (13.3 kPa). By setting the pressure within the above range, methanol (ethanol) generated when the hydroxyl group of the hydroxyapatite sintered body reacts with the alkoxysilyl group can be removed.
  • reaction conditions of the above reaction step, the type of solvent, and the like may be appropriately changed depending on the type of inorganic base material used and the type of functional group.
  • the biocompatible ceramic particles can be used without pretreatment. Therefore, an artificial blood vessel can be produced without modifying the properties of the biocompatible ceramic particles.
  • the biocompatible ceramic particles and the binder are reacted to introduce a reactive group capable of reacting with a hydroxyl group on the surface of the biocompatible ceramic particles.
  • a silane coupling agent having the alkoxysilyl group (reactive functional group) and the reactive group is used as the binder, and titanium oxide is used as the biocompatible ceramic particle. I will explain.
  • reaction conditions for reacting the alkoxysilyl group of the silane coupling agent with titanium oxide differ depending on the type of reaction, the type of silane coupling agent used, and the like, and are not particularly limited. Absent. Moreover, as a kind of said reaction, a dry method, a wet method, etc. are suitable, for example.
  • titanium oxide particles are put into a high-speed stirrer, and a silane coupling agent having a reactive group at one end and an alkoxysilyl group at the other end is dropped or sprayed thereon. After adding and stirring uniformly, it may be dried. At this time, the addition amount of the silane coupling agent is more preferably in the range of 0.0001 to LO parts by weight with respect to 1 part by weight of the titanium oxide particles.
  • titanium oxide particles and a silane coupling agent are added to an organic solvent, and the mixture is stirred within a temperature range of room temperature to 150 ° C. After reacting for minutes to 10 days, the solvent and unreacted silane coupling agent may be removed and dried.
  • organic solvent used here include hydrocarbon solvents such as toluene and hexane; ether solvents such as tetrahydrofuran and jetyl ether; ketone solvents such as acetone and methyl ethyl ketone; It is done.
  • the lower limit of the amount of the organic solvent used is preferably 0.1 parts by weight or more, more preferably 0.5 parts by weight or more, per 1 part by weight of the acid-titanium particles (acid-titanium particles). Further preferred. When the amount of the organic solvent used is less than 0.1 parts by weight, the surface of the acid titanium particles, which are difficult to make the reaction system uniform, may not be uniformly modified.
  • the upper limit of the amount of the organic solvent used is more preferably 1000 parts by weight or less, more preferably 50 parts by weight or less, with respect to 1 part by weight of titanium oxide particles. When the amount of the organic solvent used is more than 1000 parts by weight, it is not economical.
  • the lower limit of the addition amount of the silane coupling agent is more preferably 0.0001 parts by weight or more and further preferably 0.001 parts by weight or more with respect to 1 part by weight of the titanium oxide particles.
  • the addition amount of the silane force pulling agent is less than 0.0001 parts by weight, the amount of reactive groups introduced into the surface of the titanium oxide particles may not be sufficient.
  • the upper limit of the addition amount of the silane coupling agent is more preferably 10 parts by weight or less, more preferably 5 parts by weight or less, with respect to 1 part by weight of oxytitanium particles. It is not economical when the amount of silane coupling agent added is more than 10 parts by weight. The reason for this is the same in the case of the dry method.
  • the lower limit of the reaction temperature is more preferably room temperature (25 ° C) or higher. When the reaction temperature is lower than the room temperature, the reaction may take a long time.
  • the upper limit of the reaction temperature is more preferably 100 ° C. or less, more preferably 150 ° C. or less. When the reaction temperature is higher than 150 ° C, the alkoxysilyl group and Z or the reactive group at the terminal of the silane coupling agent may cause an undesirable side reaction.
  • the reaction temperature is preferably about 80 ° C.
  • the reaction step the hydroxyl group generated on the surface of the artificial blood vessel substrate by the hydroxyl group generation step is reacted with the reactive group introduced into titanium oxide by the reactive group introduction step. to this Thus, an artificial blood vessel in which the biocompatible ceramic particles and the artificial blood vessel substrate are bonded by chemical bonding can be produced.
  • the specific reaction conditions are the same as those in the direct reaction step described above, and detailed description thereof is omitted.
  • the artificial blood vessel that is useful in the present embodiment has a configuration in which biocompatible ceramic particles are chemically bonded to the surface of the artificial blood vessel substrate.
  • the lower limit of the coverage of the biocompatible ceramic particles on the artificial blood vessel substrate is 10% or more, more preferably 20%, assuming that the surface area of the artificial blood vessel substrate is 100%. More preferably 25% or more. When the coverage is less than 10%, vascular endothelial cells may not adhere sufficiently to the artificial blood vessel surface.
  • the coverage is 100% or more, that is, a structure in which biocompatible ceramic particles are further laminated on the biocompatible ceramic particles bonded to the artificial blood vessel substrate. In this case, the biocompatible ceramic particles that are stacked are not chemically bonded to the artificial blood vessel base material and are only physically adsorbed, and may be detached. For this reason, the artificial blood vessel is particularly preferred that the biocompatible ceramic particles are chemically bonded to the artificial blood vessel substrate in a single layer!
  • the thickness of the biocompatible ceramic particle layer in the obtained artificial blood vessel is different depending on the force depending on the thickness of the human blood vessel substrate, for example, when the artificial blood vessel substrate thickness is 100%.
  • the range of 0001% to 100% is more preferable.
  • the range of 0.001% to 10% is more preferable.
  • polyester-based resin and Z or polyurethane-based resin are used as the artificial blood vessel base material, and after these artificial blood vessel base materials are hydrolyzed, biocompatible ceramic particles are bound. I am letting.
  • the material constituting the artificial blood vessel base material include only the polyester-based resin and Z or polyurethane-based resin.
  • a fluoride of polyethylene such as polytetrafluoroethylene may be used.
  • polyethylene fluoride is used as the human blood vessel substrate and is hydrolyzed, the end of the polyethylene fluoride is hydrolyzed, and a hydroxyl group is generated at the end. . And using the hydroxyl group produced
  • the artificial blood vessel may be produced by performing any one of (1) a reactive functional group introduction step and a direct reaction step, and (2) a reactive group introduction step and a reaction step.
  • the reaction was conducted at a reaction temperature of 50 ° C. with stirring for 24 hours.
  • the resulting reaction product was separated by centrifugation to obtain nodyloxyapatite.
  • the hydroxyapatite was heated at 800 ° C. for 1 hour to obtain hydroxyapatite sintered particles (hereinafter referred to as “HAp particles”). These HAp particles are single crystals.
  • the major axis force was Sl50 to 400 nm.
  • the hydrolyzed artificial blood vessel substrate was washed with a large amount of pure water, and then the artificial blood vessel substrate was dried at 60 ° C.
  • the hydrolyzed artificial blood vessel substrate was put into a three-necked flask under a nitrogen atmosphere, and 45 ml of benzyl alcohol as a polymerization solvent and ⁇ -methacryloxypropyl as a silane coupling agent were added. 5 ml of triethoxysilane was added and inflated at 85 ° C for 30 minutes.
  • Table 1 shows the introduction rate of alkoxysilyl groups during the reaction time.
  • the introduction rate was determined by the following equation (1), where Ag is the weight of the untreated artificial blood vessel substrate and Bg is the weight of the artificial blood vessel substrate after introduction of the hydroxyl group.
  • nodyl and hydroxyapatite sintered particles produced as described above were added in ethanol. Then, a dispersion liquid was prepared so as to be 2% by weight.
  • the hydroxyapatite sintered body particles were adsorbed on the surface of the artificial blood vessel substrate by immersing the artificial blood vessel substrate into which the alkoxysilyl group was introduced in the dispersion for 1 hour. Thereafter, the artificial blood vessel substrate was taken out and washed several times with ethanol.
  • the hydroxypatite sintered body was adsorbed to react the hydroxyapatite sintered body adsorbed on the surface of the artificial blood vessel substrate with the alkoxysilyl group introduced into the artificial blood vessel substrate.
  • the artificial blood vessel substrate was subjected to a coupling reaction at 80 ° C for 2 hours.
  • the obtained reaction product was immersed in ethanol and irradiated with ultrasonic waves for 1 minute to remove unreacted hydroxyapatite sintered particles and obtain an artificial blood vessel according to the present invention.
  • the obtained artificial blood vessel was observed with a scanning electron microscope. The results are shown in Fig. 1.
  • the artificial blood vessel placed in a 24-multiwell dish, the artificial blood vessel made of polyester used as the artificial blood vessel base material (hereinafter referred to as “comparative artificial blood vessel” in this example), and the comparative artificial blood vessel Human umbilical vein endothelial cells (H UVEC) were seeded at a rate of 1 ⁇ 10 5 cells / ml, respectively, on collagen-coated surfaces (hereinafter referred to as “collagen-coated artificial blood vessels” in this example). .
  • the comparative artificial blood vessel and the collagen-coated artificial blood vessel correspond to comparative examples.
  • EBM-2 bovine serum, gentamicin Z amphotericin B, hydrocortisone, hFGF-B, VEGF, R3-IGF-1, ascorbic acid, hEGF
  • the cell adhesiveness is remarkably improved as compared with the comparative example (comparative artificial blood vessel), and compared with the collagen-coated artificial blood vessel (collagen-coated artificial blood vessel). It can be seen that there is no difference.
  • the artificial blood vessel substrate As the artificial blood vessel substrate, a polyester artificial blood vessel was used. Corona discharge treatment was applied to the artificial blood vessel substrate at a voltage of 100 V for 12 seconds. Thereafter, the artificial blood vessel substrate subjected to the corona discharge treatment and HEMA were subjected to graft polymerization by reacting at 60 ° C. for 60 minutes under normal atmospheric pressure conditions. Thereafter, the graft-polymerized artificial blood vessel substrate was sonicated in ethanol for 1 minute. Thereafter, the vascular substrate after the ultrasonic treatment was washed with ethanol for 2 hours and then dried. As a result, an artificial blood vessel substrate in which a hydroxyl group was generated was obtained (the dried artificial blood vessel substrate is hereinafter referred to as “HEMA human blood vessel substrate”).
  • Fig. 5 shows the formation of the hydroxyl group confirmed by FT-IR ATR.
  • (a) shows the results of the comparative artificial blood vessel as a comparative example, and (b) shows the HEMA artificial blood vessel substrate.
  • the deoxygenated reaction solution was heated in a 60 ° C hot water bath for 60 minutes. Thereafter, in order to remove unreacted monomers and homopolymer, the artificial blood vessel substrate was taken out of the heated reaction solution and washed for 2 hours with stirring in ethanol (this artificial blood vessel and the following and In the figure, it is called “KBE artificial blood vessel substrate”).
  • Figure 7 shows the results of confirming the introduction of alkoxysilyl groups by FT-IR ATR.
  • FIG. 7 shows the results of the comparative artificial blood vessel as a comparative example
  • (b) shows the results of the KBE artificial blood vessel substrate
  • (c) shows ⁇ -methacryloxypropyltriethoxysilane ( ⁇ 503). ) Shows the results of the monomer.
  • the graft polymer introduction rate at this time was about 2.2%.
  • the graft polymer introduction ratio is calculated by the following formula.
  • the weight of the artificial blood vessel substrate was determined as Dg, and the weight of the KBE artificial blood vessel substrate as Eg.
  • Figure 8 shows the results of observation of the surface of the artificial blood vessel substrate with a scanning electron microscope.
  • the hydroxyapatite sintered particles according to this example were produced in the same manner as the method shown in Example 1.
  • a dispersion liquid was prepared by dispersing the produced hydroxyapatite sintered body particles in ethanol so that the particle weight would be 2% by weight.
  • the KBE artificial blood vessel base material was placed in the dispersion and stirred for 1 hour, thereby adsorbing the hydrite oxy-vertate sintered body particles to the KBE artificial blood vessel base material. Thereafter, the artificial blood vessel substrate adsorbed with the above-mentioned sintered particles of idroxiapatite was taken out and washed several times with ethanol.
  • EBM-2 bovine serum, gentamicin / amphotericin B, hide cortisone, hFGF-B, VEGF, R3-IGF-1, ascorbic acid, hEGF was used for 4 hours as a culture solution. It was.
  • FIG. 10 shows the results of observation of cell adhesion of the above three samples with a scanning electron microscope.
  • FIG. 10 shows a scanning electron microscope image of an artificial blood vessel according to the present invention
  • (b) shows a scanning electron microscope image of a collagen-coated artificial blood vessel as a comparative example
  • (c) shows a comparative example.
  • the scanning electron microscope image of the comparative artificial blood vessel which is is shown.
  • FIG. 11 shows the appearance of the apatite-coated artificial blood vessel used in this example and an operation electron microscope image.
  • FIG. 11 shows the appearance of the artificial blood vessel according to the present invention, and (b) shows a scanning electron microscope image of the surface of the artificial blood vessel.
  • Example 2 an artificial blood vessel before being combined with the hydroxyapatite sintered body particles, which was referred to as KBE artificial blood vessel base material, was used as a comparative example (in this example, “untreated” hereinafter). Referred to as a "artificial blood vessel”). In addition, 0.1% collagen (trade name: Cellmatrix I-C, Nitta Gelatin Co., Ltd.) is coated on a polyester artificial blood vessel sterilized with ethylene oxide gas (hereinafter referred to as “EOG sterilization”). An artificial blood vessel was prepared (hereinafter referred to as “collagen-coated artificial blood vessel” in this example).
  • EOG sterilization ethylene oxide gas
  • Diazepam injection solution 10mgZ2ml
  • Attosulfate sulfate injection solution (Attosulfate sulfate) lmgZ2ml
  • Ketamine hydrochloride ketalal 10mgZkg
  • injection Ampicillin sodium (bicillin for injection) lgZ4ml
  • S-in isoflurane
  • FIG. 12 shows a state before and after the left and right carotid arteries are replaced with the untreated artificial blood vessel and the apatite-coated artificial blood vessel.
  • Fig. 12 shows the blood vessel before the untreated artificial blood vessel is implanted
  • c shows the blood vessel before the apatite coating artificial blood vessel is implanted
  • b shows the untreated blood vessel.
  • D shows a blood vessel in which an apatite-coated artificial blood vessel is implanted.
  • FIG. 12 (e) is a drawing showing the appearance of the entire implanted blood vessel.
  • the upper blood vessel is an untreated artificial blood vessel implanted
  • the lower blood vessel is an apatite-coated human.
  • the blood vessel is implanted.
  • the muscle and the subcutaneous are continuously sutured with an absorbable suture (trade name: Vicryl, USP No. 4-0, manufactured by ETHI CON), and the skin is nylon thread (trade name: EIP, USP No. 2-0, manufactured by Akiyama Mfg. Co., Ltd.).
  • an absorbable suture trade name: Vicryl, USP No. 4-0, manufactured by ETHI CON
  • the skin is nylon thread
  • EIP trade name: EIP, USP No. 2-0, manufactured by Akiyama Mfg. Co., Ltd.
  • HE staining is a method for staining cell nuclei and cytoplasm, which is simply used to observe the morphology of a tissue, and is the most common tissue staining method commonly used for most tissues.
  • liquid A hematoxylin l.Og, ethanol 10 ml
  • liquid B potassium iodoban 50 g, ultrapure water 800 ml
  • the liquid A, the liquid B, and 0.2 g of sodium iodate were mixed and allowed to stand for 1 hour. Thereafter, 200 ml of glycerin was mixed.
  • this mixed solution is simply referred to as “hematoxylin”.
  • the Eosin solution used for HE staining is 100 ml of water, Yejin Y 1. Og, The mixture was stored as a stock solution, and the stock solution was diluted 4 times with 80% ethanol at the time of use, and 10 ml of 10% acetic acid was mixed and used. It should be noted that the amount of Ellis Mouth Syn may be appropriately adjusted according to the result of staining.
  • Paraffin sections of tissue samples are placed in a staining basket and then immersed in xylene for 5 minutes. This was repeated once more, and then immersed in 100%, 90%, 80%, 70% and 50% ethanol stepwise for 5 minutes, 5-6 times, respectively.
  • the cells were immersed in the above-mentioned matoxylin for 5 minutes. Thereafter, the tissue was washed with running water, taking care that the momentum of the water flow was strong and the part was not in direct contact with the tissue sample.
  • washed tissue sample was immersed in the eosin solution for 5 minutes 5 to 6 times, and then washed with water.
  • tissue sample is immersed in xylene for 5 minutes for 5 times. This was further repeated by repeating this process four times.
  • Paraffin sections of tissue samples were dried at 37 ° C. In addition, activation treatment is not performed. The tissue sample was deparaffinized and then washed with water. The deparaffinization method was the same as that used in the HE staining.
  • tissue sample 80% methanol, and 0.6% hydrogen peroxide were mixed and reacted for 15 minutes.
  • tissue sample after washing with water, the tissue sample and 3% hydrogen peroxide were mixed and reacted for 15 minutes.
  • the tissue sample was immersed in TBST (50 mM Tris-HCU pH 7.6, 0.15 M NaCl, 0.05% Tween) for 5 minutes.
  • a tissue sample and an anti-human smooth muscle fiber diluted 100-fold were used as a primary antibody reaction. It was reacted with a cutin ⁇ 1-4 mouse monoclonal antibody (DAKO, code NO .: M0841) at room temperature for 30 minutes. After the primary antibody reaction, it was washed with the above TBST.
  • DAKO cutin ⁇ 1-4 mouse monoclonal antibody
  • ENVISION staining method was performed by ENVISION staining method.
  • ENVISION staining system ENVISION + ZHRP for mouse primary antibody> (DAKO, code NO .: M4001) was used. At this time, the reaction temperature was room temperature and the reaction time was 30 minutes. Then, it wash
  • the tissue sample was developed with dimethylaminoazobenzene (DAB), washed with water, and then nuclear-stained with the above hematoxylin. After nuclear staining, the tissue samples were colored by washing with water.
  • DAB dimethylaminoazobenzene
  • a-SMA staining was performed according to the above VW staining. However, it differs from VW staining in the following points.
  • tissue sample was washed with water and then used with proteinase K. 22.
  • Antigen activation treatment was performed at 5 ° C for 1 minute.
  • the anti-human smooth muscle fiber lactin '1-4 mouse monoclonal antibody was diluted 500-fold.
  • the coloration after the nuclear staining was powerful.
  • FIG. 13 shows the results of the above evaluation two weeks after implantation.
  • (a) is a drawing showing the appearance of the untreated artificial blood vessel cut
  • (b) is an enlarged view of the vicinity of the anastomosis portion of (a)
  • (c) is a drawing of (a).
  • 2 is a drawing showing a scanning electron microscope image of the surface of the anastomosis.
  • (D) is a drawing of a cut untreated artificial blood vessel subjected to HE staining
  • (e) is an enlarged view of the intima side of the blood vessel in the part enclosed by a solid line in (d).
  • (F) is an enlarged view of the adventitia side of the blood vessel in the part surrounded by a solid line in (d).
  • (G) is a drawing of a cut untreated artificial blood vessel subjected to VW staining
  • (h) is an enlarged view of the intima side of the blood vessel in the part surrounded by a solid line in (g).
  • (I) is an enlarged view of the outer membrane side of the blood vessel in the part surrounded by a solid line in (g).
  • (J) Is a drawing showing the appearance of an apatite-coated artificial blood vessel cut, and (h) is (j
  • (m) is a drawing of a cut apatite-coated artificial blood vessel subjected to HE staining
  • (n) is an enlarged view of the intima side of the blood vessel in the portion surrounded by a solid line in (m).
  • O) is an enlarged view of the adventitia side of the blood vessel in the part surrounded by a solid line in (m).
  • (P) is a drawing of a cut apatite-coated artificial blood vessel subjected to VW staining
  • (q) is an enlarged view of the intima side of the blood vessel in the part surrounded by a solid line in (p).
  • (R) is an enlarged view of the outer membrane side of the blood vessel in the part surrounded by a solid line in (p).
  • FIG. 14 shows the results of the evaluation of implanted plants after 4 weeks.
  • (a) is a drawing showing the appearance of a cut untreated artificial blood vessel
  • (b) is an enlarged view of the vicinity of the anastomosis part of (a)
  • (c) is a drawing of (a).
  • 2 is a drawing showing a scanning electron microscope image of the surface of the anastomosis.
  • (D) is a drawing of a cut untreated artificial blood vessel subjected to HE staining
  • (e) is an enlarged view of the intima side of the blood vessel in the part enclosed by a solid line in (d).
  • (F) is an enlarged view of the adventitia side of the blood vessel in the part surrounded by a solid line in (d).
  • (G) is a drawing of a cut untreated artificial blood vessel subjected to VW staining, and (h) is an enlarged view of the intima side of the blood vessel in the part surrounded by a solid line in (g).
  • (I) is an enlarged view of the outer membrane side of the blood vessel in the part surrounded by a solid line in (g).
  • (J) is a drawing showing the appearance of the apatite-coated artificial blood vessel cut, (h) is an enlarged view of the vicinity of the anastomosis part of (j), and (1) is the surface of the anastomosis part of (j).
  • (m) is a drawing of the cut apatite-coated artificial blood vessel for HE staining.
  • N is an enlarged view of the intima side of the blood vessel in the part enclosed by the solid line in (m), and (o) is enclosed by the solid line in (m). It is the figure which expanded the adventitial side of the blood vessel among the parts.
  • P is a drawing of a cut apatite-coated artificial blood vessel subjected to VW staining
  • (q) is an enlarged view of the intima side of the blood vessel in the part surrounded by a solid line in (p).
  • R is an enlarged view of the outer membrane side of the blood vessel in the part surrounded by a solid line in (p).
  • Fig. 15 shows the results of comparison of the extension distance of vascular endothelial cells into the test artificial blood vessel.
  • the vertical axis indicates the length of extension of vascular endothelial cells
  • the unit is millimeters
  • the horizontal axis indicates the time that has elapsed since implantation, the unit is weeks
  • (a) The bar indicated by represents the length of the vascular endothelial cells in the untreated artificial blood vessel
  • the bar indicated by (b) represents the length of the vascular endothelial cells in the apatite-coated artificial blood vessel.
  • the apatite-coated artificial blood vessel had a longer vascular endothelial cell extension distance than the untreated artificial blood vessel. From this result, it was found that an apatite-coated artificial blood vessel is suitable as a cell scaffold.
  • FIG. 16 shows the result of comparison of the thickness of the thrombus formed in the test artificial blood vessel.
  • FIG. 16 it is a drawing showing the result of comparing the thickness of the thrombus formed in the artificial blood vessel after implantation, the vertical axis shows the thickness of the vascular endothelial cell extended, the unit is micrometer, The horizontal axis shows the time elapsed since implantation, the unit is week, the bar shown in (a) represents the thickness of the thrombus formed in the untreated artificial blood vessel, ( The bar indicated by b) represents the thickness of the thrombus formed in the apatite-coated artificial blood vessel.
  • the apatite-coated artificial blood vessel has an anchoring effect that thrombus detachment is suppressed and the extracellular matrix enters the interface of the blood vessel substrate.
  • FIG. 18 shows the results after 4 weeks for implanted plants.
  • FIG. 17 (a) is a drawing in which the intima side of the untreated artificial blood vessel is enlarged 200 times, and (b) is a drawing in which the outer membrane side is enlarged 200 times.
  • (C) is a drawing in which the inner membrane side of the apatite-coated human blood vessel is enlarged 200 times,
  • (b) is a drawing in which the outer membrane side is enlarged 200 times, and arrow (i) is the inner drawing. This is the fiber of the artificial blood vessel on the membrane side, and (ii) is the fiber of the artificial blood vessel on the outer membrane side.
  • FIG. 18 is a view in which the intima side of an untreated artificial blood vessel is enlarged 100 times, and (b) is a circle with a solid line in (a). It is a drawing in which the enclosed part is enlarged 200 times, (c) is a drawing in which the outer membrane side is enlarged 200 times in the untreated artificial blood vessel, and (d) is a drawing in which the untreated artificial blood vessel is enlarged. (E) is a drawing in which the part surrounded by a solid line in a circle is magnified 200 times in (d), and (f) is an undrawn drawing. Treated artificial blood vessels In the figure, the outer membrane side is magnified 200 times, the arrow (i) is the fiber of the artificial blood vessel on the inner membrane side, and (ii) is the fiber of the artificial blood vessel on the outer membrane side.
  • the untreated artificial blood vessel when the untreated artificial blood vessel is compared with the apatite-coated artificial blood vessel, the untreated artificial blood vessel has more round cells. This indicates immature smooth muscle cells. However, in the case of an apatite-coated artificial blood vessel, there are many spindle-shaped cells. This indicates that there are many mature smooth muscle cells.
  • FIG. 19 shows the results of implanting a collagen-coated artificial blood vessel and an apatite-coated artificial blood vessel.
  • (a) is a drawing showing the appearance of the apatite-coated artificial blood vessel cut
  • (b) is a drawing showing the appearance of the collagen-coated artificial blood vessel cut.
  • the artificial blood vessel according to the present invention is suitable as a medium-diameter artificial blood vessel having an inner diameter of 5 mm, which can be achieved by a conventionally known artificial blood vessel.
  • the artificial blood vessel of the present invention is particularly suitable for use for long-term implantation in a living body.
  • the present invention is particularly useful in the medical device industry.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Transplantation (AREA)
  • Epidemiology (AREA)
  • Veterinary Medicine (AREA)
  • Dermatology (AREA)
  • Medicinal Chemistry (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Materials Engineering (AREA)
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  • Prostheses (AREA)

Abstract

Vaisseau sanguin synthétique qui ne fait pas usage de matières dérivées d'organismes vivants et présente une biocompatibilité si élevée que les cellules endothéliales vasculaires peuvent se développer à un stade précoce dans le vaisseau sanguin synthétique. Le vaisseau sanguin synthétique est produit par un procédé servant à produire un vaisseau sanguin synthétique composé d'une matière de base de vaisseau sanguin synthétique comprenant au moins une matière sélectionnée entre une résine de polyester, une résine de polyuréthane et une résine fluorée et, liées chimiquement à celle-ci, des particules de céramique biocompatible ayant une biocompatibilité appropriée, lequel procédé comprend une étape d'hydroxylation consistant à hydroxyler une matière de base de vaisseau sanguin synthétique, ou alternativement après un traitement de surface, effectuer une polymérisation par greffage d'un ester (méth)acrylique hydroxylé ou similaire sur celle-ci, de façon à pourvoir des hydroxyles sur la surface de la matière de base de vaisseau sanguin synthétique ; une étape d'introduction d'un groupe fonctionnel réactif consistant à faire en sorte qu'un liant ayant un groupe réactif capable de se lier chimiquement aux hydroxyles et un groupe fonctionnel réactif capable de se lier chimiquement aux particules de céramique biocompatible réagisse avec les hydroxyles de la matière de base de vaisseau sanguin synthétique de façon à introduire le groupe fonctionnel réactif dans la matière de base de vaisseau sanguin synthétique ; et une étape de réaction directe consistant à faire réagir le groupe fonctionnel réactif avec les particules de céramique biocompatible.
PCT/JP2006/308293 2005-07-12 2006-04-20 Vaisseau sanguin synthétique et procédé servant à produire celui-ci WO2007007452A1 (fr)

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Publication number Priority date Publication date Assignee Title
US8360000B2 (en) 2005-11-28 2013-01-29 Japan Science And Technology Agency Flocked medical instrument to be placed in the body, method of producing the medical instrument to be placed in the body and apparatus for producing the medical instrument to be placed in the body
JPWO2018043620A1 (ja) * 2016-08-30 2019-07-04 株式会社ソフセラ セラミック粒子担持医療用チューブ及び／又はカフ
JP2020005995A (ja) * 2018-07-10 2020-01-16 帝人ナカシマメディカル株式会社 硬組織インプラント
JP7333552B2 (ja) 2018-07-10 2023-08-25 帝人ナカシマメディカル株式会社 硬組織インプラント

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