CN106963988A - 一种嵌合型组织工程牛心包片 - Google Patents

一种嵌合型组织工程牛心包片 Download PDF

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CN106963988A
CN106963988A CN201610021779.4A CN201610021779A CN106963988A CN 106963988 A CN106963988 A CN 106963988A CN 201610021779 A CN201610021779 A CN 201610021779A CN 106963988 A CN106963988 A CN 106963988A
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cell
bovine pericardium
organizational project
mosaic type
pericardium
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李温斌
辛志飞
万居易
李殿坤
马小龙
龚达
张燕
许秀芳
辛毅
周子凡
徐志伟
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    • 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
    • A61L27/3683—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
    • A61L27/40—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
    • A61L27/48—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix with macromolecular fillers
    • 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
    • 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

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  • Health & Medical Sciences (AREA)
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  • Animal Behavior & Ethology (AREA)
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  • Veterinary Medicine (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Epidemiology (AREA)
  • Transplantation (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Medicinal Chemistry (AREA)
  • Dermatology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
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  • Vascular Medicine (AREA)
  • Urology & Nephrology (AREA)
  • Zoology (AREA)
  • Cardiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • General Chemical & Material Sciences (AREA)
  • Composite Materials (AREA)
  • Materials Engineering (AREA)
  • Materials For Medical Uses (AREA)
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Abstract

一种嵌合型组织工程牛心包片。牛心包中层是由细胞、基质、纤维三部分组成。纤维维持牛心包生物力学特性，细胞存在于纤维空隙中，通过基质成份与纤维形成连接。临床常用戊二醛处理的牛心包植入体内产生钙化、血栓形成和衰败的原因如下：1.戊二醛固定方法能一定程度上降低免疫原性，但仍有异种细胞成分残留，具有细胞毒性，可诱发免疫排异反应，进一步引起组织钙化、衰败；2，中层空隙较小，机体细胞难以长入，并且诱发排斥反应的表位暴露；3，残留的醛基有细胞毒性，细胞很难存活，再生能力较低。为有效解决上述问题，在去细胞基础上再行类基质填充、交联，形成嵌合型组织工程牛心包片。

Description

一种嵌合型组织工程牛心包片
技术领域
医学 组织工程学。
背景技术
瓣膜置换术是治疗严重心脏瓣膜疾病的有效手段，目前临床使用的生物瓣膜多为戊二醛处理的猪生物瓣和牛心包瓣，因其具有良好的血流动力学特性，组织相容性好，抗凝时间短，可减少血栓栓塞或出血并发症及长期口服抗凝药带来的不便，在临床上应用日益广泛。然而，与之相随的是老化衰败及钙化问题，大大制约了其远期效益。组织工程的兴起为心血管补片、人工心脏瓣膜的研制提供了新的思路。
正常牛心包的中层是由细胞、基质、纤维三部分组成。纤维是维持牛心包生物力学特性的主要因素，细胞存在于纤维空隙中，其周围借基质成份与纤维形成连接。基质在这里指细胞和纤维以外的结构，主要由蛋白多糖和糖蛋白组成。经过对牛心包组织结构的深入研究，结果证实导致临床上常用戊二醛处理的牛心包瓣膜植入体内产生排异反应、钙化、血栓形成和力学性能不满意的原因如下：1.传统的戊二醛固定方法虽能一定程度上能降低免疫源性，但仍有异种细胞成分残留，具有细胞毒性，可诱发免疫排异反应的发生，进一步引起组织钙化、衰败；2，中层空隙较小，机体细胞难以长入，并且诱发排斥反应的表位暴露；3，残留的醛基有细胞毒性，机体的细胞很难在戊二醛处理后的牛心包内存活有，再生能力较低。为有效解决上述问题，我们在去细胞基础上再行类基质填充、交联，形成“嵌合型”组织工程牛心包片。
发明内容
嵌合型组织工程牛心包片，可作为制备人工心脏生物瓣膜的理想材料。
附图说明 图1：一种嵌合型组织工程牛心包片结构示意图（脱细胞-类基质填充-交联示意图）
具体实施方式 采用Triton X-100＋脱氧胆酸钠盐＋EDTA去细胞方法，在彻底去除组织中细胞的同时也能够最大程度地去除组织中的基质成份，仅保留牛心包片内的胶原纤维、弹力纤维，维持肺动脉支架的完整性，然后用类基质明胶进行填充，最后用EDC/NHS交联，得到“嵌合型”组织工程牛心包片。
基本步骤：将新鲜牛心包片充分修剪后，置于 0.25% Triton X-100+0.25%脱氧胆酸钠盐+0.02% EDTA的去细胞溶液中，恒温恒速（37℃，200r/min）振荡 24小时去细胞处理；去细胞结束后置于1%青链霉素+2ug/ml两性霉素B的灭菌PBS中，恒温恒速（37℃，200r/min）振荡 24小时充分漂洗，每4小时换液一次；将去细胞的牛心包片逐级降温-80℃冻存，然后置于冷冻干燥机中脱水处理24小时；将脱水处理后的牛心包片置于经紫外线照射灭菌的2%的明胶溶液中，恒温恒速（37℃，200r/min）振荡24小时，然后将明胶嵌合的牛心包片置于1%青链霉素+2ug/ml两性霉素B的灭菌PBS中，逐级降温至-80℃冻存，然后置于冷冻干燥机中脱水处理24小时；将脱水处理后的牛心包片置于EDC + NHS中，4℃下静置交联24小时，然后用0.1mol/L 磷酸氢钠清洗2小时，终止交联，最后用灭菌 PBS 溶液充分漂洗，得到“嵌合型”组织工程牛心包片。

Claims (1)

1.该项申请的嵌合型组织工程牛心包片，是将异种动物（牛）心包通过脱细胞-类基质填充-交联方法得到，该嵌合型组织工程牛心包片可作为制备人工心脏生物瓣膜的理想材料，应用价值较大，故要求以下专利权：（1）其他各种动物心包组织通过脱细胞-类基质填充-交联得到各种类型的组织工程心包；（2）各种脱细胞-类基质填充-交联的方法，包括应用各种浓度的Triton X-100＋脱氧胆酸钠盐＋EDTA、明胶、EDC/NHS及其他相关试剂等；（3）通过此方法及类似方法制作的心包片用于制作生物瓣膜、补片等所有移植物。
CN201610021779.4A 2016-01-14 2016-01-14 一种嵌合型组织工程牛心包片 Pending CN106963988A (zh)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019100454A1 (zh) * 2017-11-27 2019-05-31 大连理工大学 一种三维肿瘤模型脱细胞多孔支架、构建方法及其应用
CN110507856A (zh) * 2019-09-29 2019-11-29 四川大学 一种全预装干燥介入主动脉瓣膜材料及其制备方法
CN113198045A (zh) * 2021-04-29 2021-08-03 武汉纺织大学 一种贴合型生物瓣膜及其制备方法

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019100454A1 (zh) * 2017-11-27 2019-05-31 大连理工大学 一种三维肿瘤模型脱细胞多孔支架、构建方法及其应用
CN110507856A (zh) * 2019-09-29 2019-11-29 四川大学 一种全预装干燥介入主动脉瓣膜材料及其制备方法
CN113198045A (zh) * 2021-04-29 2021-08-03 武汉纺织大学 一种贴合型生物瓣膜及其制备方法

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Application publication date: 20170721