牛心包组织工程心脏瓣膜支架脱细胞的研究

目的研究去污剂-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。方法分别采用去污剂-酶消化法、胰蛋白酶消化法和去氧胆酸钠法,处理新鲜牛心包组织,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,与去污剂-酶消化法比较,其他2种方法对基质的破坏明显。结论去污剂-酶消化法脱细胞效果好,且具有良好的保护基质能力。

Study of Decellularization of Bovine Pericardium Tissue Engineered Heart Valves Scaffold

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-engineered heart valves(TEHVs). In this study different decellularization procedures were tested for their potential of cell removal and ability to preserve the matrix. Methods Specimens of bovine pericardiums were treated by detergent enzyme extraction, 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. Von Gieson(VG) staining and Gomori staining were used to evaluate 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 Complete decellularization was achieved in all 3 groups. How- ever, trypsin, Triton-X 100 and sodium-deoxycholate treatment 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 achieve both complete decellularization and preservation of the matrix structure.it might be an ideal approach to the construction of TEHVs.