牛心包组织工程心脏瓣膜支架脱细胞方法的比较  

Comparison of decellularizative procedures of bovine pericardium tissue-engineering heart valve scaffold

作　　者：杨岷[1] 陈长志[1] 成少飞[1] 王学宁[1] 叶清[1] 王维俊[1] 王小妹[1] 

机构地区：[1]上海第二医科大学附属仁济医院胸心外科,200127

出　　处：《中华胸心血管外科杂志》2005年第6期349-351,共3页Chinese Journal of Thoracic and Cardiovascular Surgery

基　　金：本课题部分受教育部留学回国人员科研基金资助(03-406-01)

摘　　要：目的对比去污剂-酶消化法、胰蛋白酶消化法和去氧胆酸钠法去除新鲜牛心包组织上细胞的效果和保护基质的能力,为组织工程心脏瓣膜的构建提供较满意的平台。方法应用3种方法处理新鲜牛心包组织,用光学显微镜、扫描电镜观察脱细胞效果和胶原纤维、弹力纤维改变;用热皱缩实验、拉力测试观察基质的物理性能变化;用DNA抽提比较脱细胞前后细胞数量差异。结果 3种方法均能完全去除细胞,与去污剂-酶消化法比较,另2种方法对基质破坏明显。结论去污剂-酶消化法脱细胞效果好,且有良好的保护基质的能力。Objective Compared with different decellularization procedures for their potential of cell removal and the abihty to preserve the matrix. Methods Specimens of bovine pericardiums were treated by 3 approaches（detergent and enzyme extractian, trypsin,Triton-X 100 and sodium-deoxycholate）. Tissue samples were then observed by HE staining and scanning electron microscopy to confirm the removal of cells. Van Gieson（VG） staining and Gomori staining were used for showing the integrity of collagen and elastin. DNA content was examined by the method of DNA extraction. Tissue shrinkage temperature and mechanical properties were also studied. Results Completely deceilularization were achieved in 3 groups. While trypsin, Triton-X 100 and sodium-deoxycholate caused severe structural destruction and declined of mechanical properties of the matrix. In contrast, detergent and enzyme extraction achieved completely deceiltfiafization and effectively preservation the matrix structure. Conclusion This research demonstrated detergent and enzyme extraction could achieve both complete deceilularization and preservation of the matrix structure. This approach may provide an ideal platform for the construction of tissue-engineering heart valves.

关 键 词：组织工程 心脏瓣膜 人工 生物假体外循环 去细胞基质 组织工程心脏瓣膜 脱细胞 牛心包 胰蛋白酶消化法 支架 扫描电镜观察 

分 类 号：R318.08[医药卫生—生物医学工程]