低渗冻融联合生物酶法制备组织工程化静脉瓣脱细胞支架  

Hypotonic freeze thawing combined with biological enzyme in preparing tissue engineering venous valve scaffold

作　　者：张永珍[1,2] 刘兴[1] 张传森[2] 

机构地区：[1]山东泰山学院体育学院理论教研室,泰安271021 [2]第二军医大学人体解剖学教研室,上海200433

出　　处：《解剖学杂志》2014年第6期731-734,799,共5页Chinese Journal of Anatomy

基　　金：国家自然科学基金（30672045）

摘　　要：目的：采用低渗冻融联合生物酶法制备组织工程化静脉瓣脱细胞支架,并与先前方法制备的静脉瓣脱细胞支架比较,以期获得一种性能较好的带瓣静脉脱细胞支架材料.方法：手术获得Beagle犬带瓣静脉分为4组,对照组、脱氧胆酸钠组、Triton组和冻融＋生物酶组.处理后对各组标本行H-E染色,透射电镜观察,DAPI荧光染色,细胞相容性测试等观察.结果：3种脱细胞方法均能彻底去除细胞,DAPI荧光检测显示各组支架材料均无DNA成分残留;H-E染色及透射电镜显示,冻融＋生物酶组胶原纤维排列整齐,未见明显的胶原纤维结构改变,其他2组可见胶原纤维断裂及结构紊乱现象;与其他2组脱细胞支架材料相比,冻融＋生物酶组的组织细胞相容性好,可见内皮祖细胞在支架材料上生长良好.结论：结合渗透压改变的反复冻融加生物酶法,既可以较彻底除去带瓣膜静脉细胞成分,又保留了较完整的细胞外基质结构,具有良好的组织和细胞相容性,是较理想的带瓣膜静脉脱细胞支架制备方法.Objective：To find a better method for preparing tissue engineered venous valve scaffold,hypotonic freeze thawing and biological enzyme were used in preparing tissue engineering venous valve matrix,and were compared with previous methods of preparation of tissue engineered venous valve matrices.Methods：Beagle veins were obtained by surgical operation and divided to four groups：an untreated group,a sodium deoxycholate group,a Triton group,and a freeze thawing plus biological enzyme group.After decellularization process,each group was observed by H-E staining,scanning electron microscopy,DAPI staining and cell compatibility testing.Results：Three methods could decellularize entirely,and no DNA residues were detected by DAPI fluorescence in each scaffold material; With H-E staining and transmission electron microscope,the collagen fibers of the freeze thawing and biological enzyme group were arranged regularly,and no obvious change was found in the collagen fiber structure,while the collagen fiber in the other two groups was found in breakage and structural disorder.The tissue compatibility of the freeze thawing and biological enzyme group was better than that of the other two groups,with visible endothelial progenitor cell complex layer well grew on scaffolds.Conclusion：The method combined with thawing and biological enzyme is an ideal method for preparation of venous valve acellular scaffold,which can decellularize entirely and retain most structure of extracellular matrix with good cell compatibility and organization.

关 键 词：脱细胞支架材料 组织工程化血管静脉瓣 脱细胞 

分 类 号：R318[医药卫生—生物医学工程]