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支架材料。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;热皱缩实验、拉力测试观察基质的物理性能变化;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,但是与去污剂-酶消化法比较,其它两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 Cardiovascular tissue engineering is a novel concept to develop ideal heart valve substitutes. The preparation of scaffolds is important for the construction of tissue-engineering heart valves(TEHVs). In this study different decellularization procedures were tested for their potential of cell removal and the ability to preserve the matrix. Methods Specimens of bovine pericardiums were treated by detergent and enzyme extraction, trypsin, Triton-X 100 and sodiumdeoxycholate. Tissue samples were then observed by HE staining and scanning electron microscopy to confirm the removal of cells. Von Gieson(VG) staining and Gomori staining were used for valuating the integrity of collagen and elastin. DNA content was examined by the method of DNA extrac- tion. Tissue shrinkage temperature and mechanical properties were also studied. Results Complete decellularization were achieved in all 3 groups. However, trypsin, Triton-X 100 and sodium-deoxycholate resulted in severe structural destruction and decreased mechanical properties of the matrix. In contrast , detergent and enzyme extraction achieved complete decellularization and effective preservation of the matrix structure. Conclusion This research demonstrated detergent and enzyme extraction could achieves both complete decellularization and preservation of the matrix structure, suggesting therefore it might be an ideal approach for the construction of TEHVs.
出处
《实用临床医药杂志》
CAS
2008年第6期20-23,27,共5页
Journal of Clinical Medicine in Practice
基金
教育部留学归国人员基金(03-476)
关键词
组织工程
心脏瓣膜
牛心包
tissue-engineering
heart valve
bovine pericardium