CN111481743B - 一种抗凝血抗钙化生物材料及其制备方法 - Google Patents

一种抗凝血抗钙化生物材料及其制备方法 Download PDF

Info

Publication number
CN111481743B
CN111481743B CN202010331255.1A CN202010331255A CN111481743B CN 111481743 B CN111481743 B CN 111481743B CN 202010331255 A CN202010331255 A CN 202010331255A CN 111481743 B CN111481743 B CN 111481743B
Authority
CN
China
Prior art keywords
anticoagulant
anticalcification
biological
preparing
biological tissues
Prior art date
Legal status (The legal status 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 status listed.)
Active
Application number
CN202010331255.1A
Other languages
English (en)
Other versions
CN111481743A (zh
Inventor
王云兵
杨凡
郭高阳
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Hangzhou Qiming Medical Devices Co ltd
Original Assignee
Sichuan University
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 Sichuan University filed Critical Sichuan University
Priority to CN202010331255.1A priority Critical patent/CN111481743B/zh
Publication of CN111481743A publication Critical patent/CN111481743A/zh
Application granted granted Critical
Publication of CN111481743B publication Critical patent/CN111481743B/zh
Priority to EP21792008.1A priority patent/EP4140511A1/en
Priority to PCT/CN2021/089448 priority patent/WO2021213516A1/zh
Priority to US18/049,237 priority patent/US20230093191A1/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

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
    • A61L33/00—Antithrombogenic treatment of surgical articles, e.g. sutures, catheters, prostheses, or of articles for the manipulation or conditioning of blood; Materials for such treatment
    • A61L33/0076—Chemical modification of the substrate
    • A61L33/0088—Chemical modification of the substrate by grafting of a monomer onto the substrate
    • 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
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
    • A61L27/3633—Extracellular matrix [ECM]
    • 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
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14—Macromolecular materials
    • A61L27/16—Macromolecular materials obtained by reactions only involving carbon-to-carbon unsaturated bonds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14—Macromolecular materials
    • A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
    • A61L27/24—Collagen
    • 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
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
    • A61L27/3625—Vascular tissue, e.g. heart valves
    • 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
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3641—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the site of application in the body
    • A61L27/367—Muscle tissue, e.g. sphincter
    • 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
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
    • A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
    • 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
    • A61L27/54—Biologically active materials, e.g. therapeutic substances
    • 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
    • A61L33/00—Antithrombogenic treatment of surgical articles, e.g. sutures, catheters, prostheses, or of articles for the manipulation or conditioning of blood; Materials for such treatment
    • A61L33/0005—Use of materials characterised by their function or physical properties
    • A61L33/0011—Anticoagulant, e.g. heparin, platelet aggregation inhibitor, fibrinolytic agent, other than enzymes, attached to the substrate
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F289/00—Macromolecular compounds obtained by polymerising monomers on to macromolecular compounds not provided for in groups C08F251/00 - C08F287/00
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/069—Vascular Endothelial cells
    • 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
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/42—Anti-thrombotic agents, anticoagulants, anti-platelet agents
    • 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
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/422—Anti-atherosclerotic agents
    • 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
    • A61L2430/00—Materials or treatment for tissue regeneration
    • A61L2430/20—Materials or treatment for tissue regeneration for reconstruction of the heart, e.g. heart valves
    • 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
    • A61L2430/00—Materials or treatment for tissue regeneration
    • A61L2430/34—Materials or treatment for tissue regeneration for soft tissue reconstruction
    • 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
    • A61L2430/00—Materials or treatment for tissue regeneration
    • A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2533/00—Supports or coatings for cell culture, characterised by material
    • C12N2533/50—Proteins
    • C12N2533/54—Collagen; Gelatin
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2533/00—Supports or coatings for cell culture, characterised by material
    • C12N2533/90—Substrates of biological origin, e.g. extracellular matrix, decellularised tissue

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Dermatology (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Transplantation (AREA)
  • Botany (AREA)
  • Molecular Biology (AREA)
  • Zoology (AREA)
  • Vascular Medicine (AREA)
  • Organic Chemistry (AREA)
  • Surgery (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Biotechnology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Materials Engineering (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Cardiology (AREA)
  • Polymers & Plastics (AREA)
  • Biophysics (AREA)
  • Cell Biology (AREA)
  • General Chemical & Material Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Materials For Medical Uses (AREA)

Abstract

本发明提供了一种抗凝血抗钙化生物材料及其制备方法，制备方法包括以下步骤：在生物组织上引入可发生聚合反应的活性基团，然后再与两性离子通过自由基共聚制得。本发明在通过生物组织中引入可进行自由基聚合的活性基团，与两性离子单体发生自由基共聚，使生物组织中的胶原蛋白通过聚合物多位点交联，实现胶原纤维内和纤维间的充分交联，提高生物组织稳定性，延长生物组织的使用寿命。同时向生物组织表面引入两性离子，可提高抗凝血性能，并且可以促进生物瓣膜的原位内皮化，进而防止钙元素沉积。

Description

一种抗凝血抗钙化生物材料及其制备方法
技术领域
本发明属于医用材料技术领域，具体涉及一种抗凝血抗钙化生物材料及其制备方法。
背景技术
异种生物组织具有和人体组织相似的物理化学性质，因此被广泛用作一种天然生物材料植入人体来替代或者修复损坏的人体组织或者器官，尤其是用作软组织修复，例如人造血管、人工瓣膜、血管补片、硬脑膜补片、疝补片、防粘连膜、软组织填充物、人工皮肤和心室辅助装置等。异种生物组织不经处理直接植入人体后由于免疫排斥作用，会迅速降解，失去机械性能。现有的生物组织为了长期发挥性能，需要通过化学交联处理才能在人体内长期稳定在。目前的异种生物组织一般通过戊二醛交联固定，但是戊二醛交联的生物组织存在明显的炎症反应和交联不稳定问题，在体内长期植入会出现降解和钙化问题，导致植入组织变硬变脆，机械性能下降，失去正常的生理功能。爱德华兹的
Figure BDA0002465025430000011
技术和美敦力的
Figure BDA0002465025430000012
技术可以减少戊二醛交联生物组织的钙化，但不能彻底消除戊二醛交联带来的生物相容性差等问题。一些其他化合物例如环氧化合物、碳化二亚胺、京尼平等新型交联剂用来交联生组织，虽然可以改善生物相容性和钙化问题，但是不能该改善其血栓原性问题。当这些组织用作血液接触材料时，由于生物组织中存在大量胶原分子，促使血液凝固，无论是戊二醛交联和非戊二醛交联均不能解决生物组织用作血液接触材料时的血栓问题。
现有技术为了解决外来植入物的血栓和免疫排斥问题，常通过共价方式对植入物表面进行功能分子修饰，希望可以减少与人体的相互作用，避免凝血和免疫系统的进攻，而共价修饰则有利于维持较长时间的功能稳定性，但是共价修饰容易导致功能分子失活，并且严重依赖修饰位点和修饰密度，常常导致修饰后的表面不具有期望的抗凝血或免疫逃避效果。对于生物组织而言，其对修饰条件有较高的敏感性，现有修饰方法常常需要较为苛刻的条件，而且修饰效率较低。因此现有技术不能同时解决生物组织植入物的钙化、凝血、免疫反应和稳定性问题，或者在解决这些问题时所使用的步骤复杂，无法得到综合性能优异的生物组织。
发明内容
针对现有技术中存在的上述问题，本发明提供一种抗凝血抗钙化生物材料及其制备方法，该生物材料具有优异的综合性能，在抗凝血、抗钙化、免疫反应和稳定性方面均较优异。
为实现上述目的，本发明解决其技术问题所采用的技术方案是：
一种抗凝血抗钙化生物材料，其制备方法包括以下步骤：
在生物组织上引入可发生聚合反应的活性基团，然后再与两性离子通过自由基共聚制得；其中，两性离子单体的结构式如下：
Figure BDA0002465025430000021
Figure BDA0002465025430000031
上述结构式命名依次为：
N,N-二甲基-N-甲基丙烯酰胺基丙基-甲烷磺酸内盐；
N,N-二甲基-N-丙烯酰胺基丙基-甲烷磺酸内盐；
N,N-二甲基-N-甲基丙烯酰胺基丙基-甲烷羧酸内盐；
N,N-二甲基-N-甲基丙烯酰胺基丙基-甲烷羧酸内盐；
[2-(甲基丙烯酰基氧基)乙基]二甲基-(3-磺酸丙基)氢氧化铵(SBMA)；
[2-(丙烯酰基氧基)乙基]二甲基-(3-磺酸丙基)氢氧化铵；
3-[[2-(甲基丙烯酰氧)乙基]二甲基铵]丙酸酯(CBMA)；
3-[[2-(丙烯酰氧)乙基]二甲基铵]丙酸酯；
2-甲基丙烯酰氧乙基磷酸胆碱(MPC)；
优选两性离子为[2-(甲基丙烯酰基氧基)乙基]二甲基-(3-磺酸丙基)氢氧化铵(SBMA)、3-[[2-(甲基丙烯酰氧)乙基]二甲基铵]丙酸酯(CBMA)或2-甲基丙烯酰氧乙基磷酸胆碱(MPC)。
制备过程具体为：将生物组织浸泡于去离子水中，然后加入活性基团，使得活性基团体积浓度为3-10％，室温反应12-36h后，清洗生物组织，然后向清洗后的生物组织中加入两性离子单体溶液，使得两性离子单体终浓度为20-500mM，于35-40℃浸泡过夜，然后加入引发剂引发聚合，制得。
进一步地，加入活性基团后，使得活性基团体积浓度为4％，室温反应24h。
进一步地，向清洗后的生物组织中加入两性离子溶液，使得两性离子终浓度为500mM，于37℃浸泡过夜。
进一步地，生物组织为心包、主动脉根、主动脉瓣、肺动脉根、肺动脉瓣、腱、韧带、皮肤、硬膜、腹膜、血管、胸膜、隔膜、二尖瓣或三尖瓣。
进一步地，活性基团为甲基丙烯酸酐、丙烯酰胺、甲基丙烯酰胺、丙烯酸酯、甲基丙烯酸酯或烯丙基。
进一步地，两性离子与生物组织通过酰胺键连接，并且与生物组织有两个以上的连接位点。
进一步地，两性离子重量占抗凝血抗钙化生物材料总重量的10-30％；优选两性离子重量占抗凝血抗钙化生物材料总重量的24.6％。
进一步地，引发剂引发聚合时反应温度不超过50℃，优选为37℃。
进一步地，引发剂为热引发剂或光引发剂，热引发剂为有机过氧化物引发剂、无机过氧化物引发剂、偶氮类引发剂、氧化还原引发剂，优选为氧化还原引发剂如过硫酸铵/亚硫酸氢钠体系；光引发剂为Irgacure D-2959。
本发明提供的抗凝血抗钙化生物材料及其制备方法，具有以下有益效果：
因未经处理的生物组织植入后在酶的作用下会发生降解，因而无法维持长期的功能，本发明在通过生物组织中引入可进行自由基聚合的活性基团，与两性离子单体发生自由基共聚，使生物组织中的胶原蛋白通过聚合物多位点交联，实现胶原纤维内和纤维间的充分交联，提高生物组织稳定性，延长生物组织的使用寿命。同时向生物组织表面引入两性离子，两性离子具有阴、阳离子基团的亲水聚合物，通过其强烈的离子溶剂化和氢键作用，在生物组织表面形成牢固的水合层，其中的水分子处于高度的自由状态能够阻碍蛋白的吸附，从而阻断下游凝血路径的启动，提高抗凝血性能，并且可以促进生物瓣膜的原位内皮化，进而防止钙元素沉积。
附图说明
图1为抗血栓和抗钙化生物组织制备流程图。
图2为抗血栓和抗钙化功能化生物制备机理示意图。
图3为MPC-20生物组织的血小板吸附SEM图片。
图4为MPC-500生物组织的血小板吸附SEM图片。
图5为SBMA-20生物组织的血小板吸附SEM图片。
图6为SBMA-500生物组织的血小板吸附SEM图片。
图7为未经修饰生物组织的血小板吸附SEM图片。
图8为戊二醛交联生物组织的血小板吸附SEM图片。
图9为内皮细胞在SBMA-500生物组织上生长荧光图片。
图10为内皮细胞在未经修饰生物组织上生长荧光图片。
图11为内皮细胞在戊二醛交联生物组织上生长荧光图片。
具体实施方式
实施例1
一种抗凝血抗钙化生物材料的制备方法，其制备流程图见图1，制备机理示意图见图2，具体步骤如下：
将新鲜猪心包用去离子水彻底清洗干净，然后加入去离子水使组织完全浸没，4℃冰浴搅拌下，逐滴加入甲基丙烯酸酐，酸酐终浓度为4％(v/v)，同时用氢氧化钠溶液维持pH值在7，滴加完毕后室温反应24h；彻底清洗甲基丙烯酸酐修饰后的猪心包，将甲基丙烯酸酐修饰后的猪心包加入如下表所示终浓度的两性离子单体溶液中，于37℃浸泡过夜，再加入引发剂交联于37℃反应24h，制得；将制备的猪心包用去离子水彻底清洗并保存在25％异丙醇溶液中备用。
未经修饰的生物组织：在加入引发剂交联前如上制备，不经单体浸泡，直接加入引发剂交联，将制备的猪心包用去离子水彻底清洗并保存在25％异丙醇溶液中备用。
具体参数如下：
Figure BDA0002465025430000061
注：SBMA:[2-(甲基丙烯酰基氧基)乙基]二甲基-(3-磺酸丙基)氢氧化铵；
MPC:2-甲基丙烯酰氧乙基磷酰胆碱；
*：未经修饰作为基线，设为0％；
引发剂浓度为刚加入反应体系时的浓度。
上述功能分子量是将制得的生物材料冻干，由于功能分子MPC中含有生物材料中几乎没有的磷元素，SBMA中含有特有的硫元素，所以使用ICP-OES分析生物材料中硫元素或者磷元素的含量，即可换算成功能分子含量，从而得到功能分子在生物组织上的修饰度，便于实验条件筛选。
具体换算方式如下：
Figure BDA0002465025430000071
其中，CS(P)(mg/L)为ICP-OES测定的硫元素或磷元素在溶液中的浓度，MS(P)为硫元素或磷元素的相对原子质量，MSBMA(MPC)为功能分子的相对分子质量，W(mg)为生物材料干重。
由上表可知，本发明制得的生物材料与对照组相比，说明功能分子很好的修饰在了生物组织上，尤其是SBMA-500组，修饰度最高。
试验例1生物组织稳定性
将制得的生物材料裁剪成3-8mg的小块，用滤纸吸干表面水分，放入铝制坩埚密封，将装有样品的坩埚放入仪器中通过差式扫描量热法分析热变性温度，从而表征生物组织的稳定性，其结果如下：
样品名 热变性温度(℃)
SBMA-500 80.15
新鲜组织 68.88
由上表可知，本发明制得的生物材料比新鲜组织更具有耐高温性，说明生物组织具有较优异的稳定性。
试验例2生物组织的抗凝血性能
将制得的生物材料清洗并裁剪成合适大小，与富含血小板的血浆孵育1h，测定粘附在材料上的血小板裂解后乳酸脱氢酶(LDH)的量，以表示血小板粘附的数量，结果如下表所示，并通过SEM观察血小板在材料上的黏附情况，具体结果见图3-8。
样品名 吸光度值(490nm)
MPC-20 0.30
MPC-500 0.29
SBMA-20 0.24
SBMA-500 0.110
未经修饰 0.32
戊二醛交联 0.35
由上表可知，490nm处的吸光度值越高表示乳酸脱氢酶的含量越高，即材料上粘附的血小板数量越多。因此，由上表可知，本发明制得的生物材料的抗凝血效果要明显优于未修饰的生物组织和经戊二醛交联生物组织(将猪心包膜浸泡于0.625％体积浓度的戊二醛水溶液中，在室温、pH＝7.4条件下交联24h，然后脱水干燥)。而经SBMA修饰后的生物组织的抗凝血效果比经MPC修饰后的生物组织的抗凝血效果好，且高嫁接度的SBMA的抗凝血效果最佳。
图3-8中SEM定性观察结果与LDH定量测量结果一致。
试验例3生物组织的抗钙化和免疫反应表征
将制得的生物材料切成1平方厘米的片材，植入SD大鼠皮下90天后取出，利用ICP-OES测定样品中钙元素含量，其结果如下：
样品名 钙元素含量(g/kg)
SBMA-500 12.8
未经修饰 38.4
戊二醛交联 117.3
由上表可知，SBMA-500修饰的生物组织钙化程度明显弱于未经修饰的和戊二醛交联生物组织。
试验例4生物组织的再细胞化性能
将制得的生物材料表面接种内皮细胞，孵育3天后使用DAPI和FITC标记的鬼笔环肽染色，结果见图9-11。
由图9-11可知，SBMA-500修饰的生物组织上内皮细胞生长情况优于未经修饰和戊二醛交联生物组织。
试验例5人工血管植入
将制得的生物材料卷成直径约为2mm的管状并使用医用粘合剂粘结接口处，用滤纸吸干表面水分，称重并记录。使用血管吻合术替换长度约2cm的兔颈动脉。植入一段时间后取出观察，并用滤纸吸干取出材料表面水分，再次称重并计算增重，即为血栓生成量，结果如下：
样品名 初始重量(mg) 最终重量(mg) 血栓生成量(mg)
SBMA-500 80.9 83.6 2.7
未经修饰 78.6 262.4 183.8
戊二醛交联 82.5 295.3 212.8
由上表可知戊二醛交联生物组织和未经修饰生物组织有超过组织重量2倍的血栓生成，SBMA-500几乎无血栓生成。

Claims (9)

1.一种抗凝血抗钙化生物材料的制备方法，其特征在于，包括以下步骤：
将生物组织浸泡于去离子水中，然后加入可发生聚合反应的活性基团，使得活性基团体积浓度为3-10％，室温反应12-36h后，清洗生物组织，然后向清洗后的生物组织中加入两性离子溶液，使得两性离子终浓度为20-500mM，于35-40℃浸泡过夜，然后加入引发剂引发聚合，制得；其中，两性离子的具体结构如下：
Figure FDA0002927246000000011
2.根据权利要求1所述的抗凝血抗钙化生物材料的制备方法，其特征在于，加入活性基团后，使得活性基团体积浓度为4％，室温反应24h。
3.根据权利要求1所述的抗凝血抗钙化生物材料的制备方法，其特征在于，向清洗后的生物组织中加入两性离子溶液，使得两性离子终浓度为500mM，于37℃浸泡过夜。
4.根据权利要求1-3任一项所述的抗凝血抗钙化生物材料的制备方法，其特征在于，生物组织为心包、主动脉根、主动脉瓣、肺动脉根、肺动脉瓣、腱、韧带、皮肤、硬膜、腹膜、血管、胸膜、隔膜、二尖瓣或三尖瓣。
5.根据权利要求1-3任一项所述的抗凝血抗钙化生物材料的制备方法，其特征在于，活性基团为甲基丙烯酸酐、丙烯酰胺、甲基丙烯酰胺、丙烯酸酯、甲基丙烯酸酯或烯丙基。
6.根据权利要求1所述的抗凝血抗钙化生物材料的制备方法，其特征在于，两性离子重量占抗凝血抗钙化生物材料总重量的1-30％。
7.根据权利要求1所述的抗凝血抗钙化生物材料的制备方法，其特征在于，引发剂引发聚合时反应温度不超过50℃。
8.根据权利要求1或7所述的抗凝血抗钙化生物材料的制备方法，其特征在于，引发剂为热引发剂或光引发剂。
9.采用权利要求1-8任一项所述方法制得的抗凝血抗钙化生物材料。
CN202010331255.1A 2020-04-24 2020-04-24 一种抗凝血抗钙化生物材料及其制备方法 Active CN111481743B (zh)

Priority Applications (4)

Application Number Priority Date Filing Date Title
CN202010331255.1A CN111481743B (zh) 2020-04-24 2020-04-24 一种抗凝血抗钙化生物材料及其制备方法
EP21792008.1A EP4140511A1 (en) 2020-04-24 2021-04-23 Anticoagulation and anticalcification biological material, and preparation method therefor
PCT/CN2021/089448 WO2021213516A1 (zh) 2020-04-24 2021-04-23 一种抗凝血抗钙化生物材料及其制备方法
US18/049,237 US20230093191A1 (en) 2020-04-24 2022-10-24 Biological Material And Preparation Method Therefor

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202010331255.1A CN111481743B (zh) 2020-04-24 2020-04-24 一种抗凝血抗钙化生物材料及其制备方法

Publications (2)

Publication Number Publication Date
CN111481743A CN111481743A (zh) 2020-08-04
CN111481743B true CN111481743B (zh) 2021-03-30

Family

ID=71790582

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202010331255.1A Active CN111481743B (zh) 2020-04-24 2020-04-24 一种抗凝血抗钙化生物材料及其制备方法

Country Status (4)

Country Link
US (1) US20230093191A1 (zh)
EP (1) EP4140511A1 (zh)
CN (1) CN111481743B (zh)
WO (1) WO2021213516A1 (zh)

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111481743B (zh) * 2020-04-24 2021-03-30 四川大学 一种抗凝血抗钙化生物材料及其制备方法
CN112773936B (zh) * 2021-02-25 2022-03-25 中国科学院金属研究所 一种改性心包膜及其制备方法、人工心脏瓣膜假体
WO2023088330A1 (zh) * 2021-11-17 2023-05-25 四川大学 一种生物瓣膜材料及其制备方法和应用
CN115970060B (zh) * 2021-11-17 2024-04-30 四川大学 一种双键后交联提高生物瓣膜材料抗钙化及抗凝血性的方法
CN114748707A (zh) * 2022-02-28 2022-07-15 四川大学 一种可降解封堵器抗凝血及快速内皮化涂层及其制备方法
CN114917415B (zh) * 2022-03-21 2023-01-06 四川大学 一种用于心脏封堵器的可降解复合膜及其制备方法和应用
CN114652366B (zh) * 2022-03-21 2023-01-06 四川大学 一种心脏封堵器用可降解复合膜及其制备方法和应用
CN115845132B (zh) * 2022-10-09 2024-02-13 北京博辉瑞进生物科技有限公司 生物补片及其制备方法和用途

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SG158172A1 (en) * 2004-12-24 2010-01-29 Celxcel Pty Ltd An implantable biomaterial and a method of producing same
CN107617126A (zh) * 2017-08-11 2018-01-23 天津大学 一种多孔材料的表面修饰方法及应用
CN109820625A (zh) * 2018-09-30 2019-05-31 四川大学 一种交联的人工生物瓣膜处理方法
CN111481743B (zh) * 2020-04-24 2021-03-30 四川大学 一种抗凝血抗钙化生物材料及其制备方法

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Radical polymerization-crosslinking method for improving extracellular matrix stability in bioprosthetic heart valves with reduced potential for calcification and inflammatory response;Gaoyang Guo等;《Acta Biomaterialia》;20181014;第82卷;摘要、第45页左栏"Mterials and methods"部分 *
Surface Modification with Poly(sulfobetaine methacrylate-co-acrylic acid) To Reduce Fibrinogen Adsorption, Platelet Adhesion, and Plasma Coagulation;Wei-Hsuan Kuo等;《Biomacromolecules》;20111113;第12卷(第12期);第4349页第2.3节，第4350页第2.5节，第4355页第5节 *

Also Published As

Publication number Publication date
US20230093191A1 (en) 2023-03-23
WO2021213516A1 (zh) 2021-10-28
EP4140511A1 (en) 2023-03-01
CN111481743A (zh) 2020-08-04

Similar Documents

Publication Publication Date Title
CN111481743B (zh) 一种抗凝血抗钙化生物材料及其制备方法
US4755593A (en) Novel biomaterial of cross-linked peritoneal tissue
KR920000459B1 (ko) 다당류 유도체가 도포된 인공혈관과 그 제조방법
CN102888027B (zh) 一种细菌纤维素/胶原-壳聚糖复合材料及其制备方法
CN111569152A (zh) 一种兼具抗凝血和抗钙化性能的生物瓣膜及其制备方法
CN111494717B (zh) 一种人工生物瓣膜及其制备方法
CN111184914B (zh) 一种功能化脱细胞基质生物材料及其制备方法和应用
CN110101912B (zh) 一种含促进内皮化多肽的干燥人工生物心脏瓣膜及其制备方法
CN111420120A (zh) 一种具有抗凝血和抗钙化功能的生物瓣膜及其制备方法
JPH07275344A (ja) 軟組織用医療用材料
CN110101911B (zh) 一种含促进内皮化生长因子的干燥人工生物心脏瓣膜及其制备方法
CN114748695B (zh) 一种双键后交联提高生物瓣膜材料抗钙化及抗凝血性的方法
CN108478298A (zh) 一种含有可结合生长因子的多糖涂层的种植体及其制备方法
WO2021164626A1 (zh) 一种功能化生物基质材料及其制备方法和应用
CN109833518B (zh) 一种生物心脏瓣膜促进内皮化的方法
WO2023088330A1 (zh) 一种生物瓣膜材料及其制备方法和应用
CN1739813A (zh) 组织工程表皮替代物及其制备方法
CN103816571B (zh) 用于角膜重建的细菌纤维素增强羊膜复合材料的制备方法
CN109971015B (zh) 一种细菌纤维素膜的脱水方法及人造血管及其制备方法
CN114028617B (zh) 一种生物材料及其制备方法和应用
CN115920131A (zh) 一种兼具抗凝和抗钙化的生物瓣膜材料、医疗器械及其交联方法和用途
CN114028618A (zh) 一种基于羊膜基底膜的生物材料及其制备方法和应用
CN116036375A (zh) 一种乙烯基双环噁唑烷交联生物瓣膜及其制备方法和用途
CN114949347A (zh) 一种改性交联生物瓣膜及其制备方法和用途
CN116848132A (zh) 产生具有受控特性的人胶原蛋白结构的方法

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
TR01 Transfer of patent right

Effective date of registration: 20220824

Address after: 310000 Room 311, 3/F, Building No. 88, Jiangling Road, Binjiang District, Hangzhou City, Zhejiang Province

Patentee after: Hangzhou Qiming Medical Devices Co.,Ltd.

Address before: 610064, No. 24, south section of first ring road, Chengdu, Sichuan, Wuhou District

Patentee before: SICHUAN University

TR01 Transfer of patent right