去细胞组织工程异种瓣膜支架制备方法的比较(英文)  

Various methods of preparing acellular tissue-engineered xenogeneic valve stents

作　　者：王鹏[1] 李超[1] 张润起[1] 谈磊[1] 孔凡华[1] 

机构地区：[1]泰安市中心医院胸外科,山东省泰安市271000

出　　处：《中国组织工程研究与临床康复》2009年第16期3041-3044,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:采用何种去细胞方法以获取良好的低抗原异种生物瓣膜支架是构建组织工程瓣膜的前提。目的:通过测试不同脱细胞方法的效果及保留脱细胞支架的能力来探讨异种组织工程心脏瓣膜生物支架的最佳制备方法。设计、时间及地点:前瞻性随机对照实验,于2007-01/2008-06在泰安市中心医院中心实验室完成。材料:16个新鲜猪主动脉瓣分为4组,对照组,NaCl-SDS组、胰蛋白酶组、曲拉通组,每组4个。方法:对照组未经去细胞处理,3个实验组分别用NaCl、胰蛋白酶、Triton-100脱细胞处理。主要观察指标:各种方法处理后的猪心脏瓣膜支架大体标本结构,光学和电子显微镜观察超微结构。采用免疫组化法检测血管内皮细胞MHC-Ⅰ类抗原的表达。结果:NaCl-SDS法脱细胞处理的瓣膜内皮细胞去除不彻底,细胞外基质的三维网状结构虽完整但纤维模糊,肿胀。胰蛋白酶法处理的瓣膜内皮细胞基本去除,但瓣膜支架结构改变很明显,胶原纤维肿胀,边缘毛糙,纤维网状间隙增宽、不规则。曲拉通法处理的瓣膜内皮细胞去除彻底,支架结构保存完好。NaCl-SDS法、胰蛋白酶法和曲拉通法脱细胞后的瓣膜均存在一定的免疫原性,但用胰蛋白酶法和曲拉通法脱细胞后的瓣膜支架所表达的免疫原性较NaCl-SDS法显著降低。结论:应用曲拉通法制作的瓣膜支架去内皮细胞较为彻底,不改变瓣叶支架结构,而且免疫原性小。BACKGROUND： Excellent Iow-antigenicity xenogeneic biological valve scaffold is the premise of constructing tissue-engineered valve by using which kind of aceUular methods. OBJECTIVE： To explore the optimal preparation method of making tissue engineered heart valves by measuring efficiency of different acellular methods and ability to preserve the matrix. DESIGN, TIME AND SETTING： The prospective randomly controlled study was performed at the Central Laboratory of Taian Central Hospital from January 2007 to June 2008. MATERIALS： Sixteen specimens of porcine aortic valves were randomly divided into control, NaCI-sodium-dodecyl-sulfate （SDS） trypsin and triton-X100 groups. METHODS： Specimens in the control group were left intact. Three test groups were decellularized with NaCI, trypsin and Triton-X100 respectively. MAIN OUTCOME MEASURES： The gross structure, optical and electron microscope ultrastructure of the decelluarated porcine heart valve matrix was compared. The expression of vascular endothelial cell major histocompatibility complex （MHC）- Ⅰ antigen was detected by immunohistochemical method. RESULTS： Treatment with NaCL-SDS achieved only incomplete decellularization. The main components of extracellular matrix were reserved completely, but the fibrous components became unclear and swelling. Treatment with trypsin removed cells completely, but caused serious structural alterations, with the presence of swollen collagen fiber, crude edge, widen and irregular fiber interspace. Treatment with Triton-X100 achieved both complete decelluarization and preservation of the matrix structure. Valves following treatment of NaCI-SDS, trypsin and Triton-X100 had certain immunogenicity. However, the immunogenicity of valves following treatment of trypsin and Triton-X100 was significantly lower compared with the treatment of NaCL-SDS. CONCLUSION： The decellularization method by Triton-X100 is effective and complete. The Triton-X100 method does not change matrix structure and has low immunogenicity.

关 键 词：组织工程:心脏瓣膜 脱细胞方法 主要组织相容性复合体 

分 类 号：R318[医药卫生—生物医学工程]