脱细胞天然支架生物力学性能变化及预处理改善组织相容性的实验研究  

Experimental study on mechanical properties of decellularized porcine aortic valve and effects of precoating methods of biological scaffold on histocompatibility

作　　者：董念国[1] 叶晓峰[1] 孙宗全[1] 史嘉玮[1] 仇玉明[1] 陈家军[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院心外科,武汉430022

出　　处：《中华外科杂志》2007年第16期1128-1131,共4页Chinese Journal of Surgery

基　　金：国家自然科学基金(30371414;30571839)

摘　　要：目的观察脱细胞猪主动脉瓣的生物力学性能变化,探讨不同预处理改善天然支架组织相容性的效果。方法新鲜猪主动脉瓣经酶加去污剂法去除细胞,力学测试仪检测其最大负荷、最大应力、最大应变和弹性模量的变化,苏木精一伊红(HE)染色、Ⅰ型胶原免疫组化染色和扫描电镜观察其病理形态学变化;将脱细胞瓣膜分别予磷酸缓冲液、多聚赖氨酸和未灭活胎牛血清包被处理,然后种植大鼠主动脉肌成纤维细胞,甲基噻唑基四唑试验检测细胞黏附率,HE 染色和扫描电镜观察形态学变化。结果酶加去污剂法能完全脱去瓣膜细胞,基本维持胶原纤维的空间结构,但其最大负荷、最大应力及弹性模量下降,最大应变上升(P<0.05);胎牛血清预处理去细胞瓣能显著提高肌成纤维细胞的黏附率,促进细胞生长、分化和增殖,并在瓣膜表面形成连续的细胞层(F 值=129.26,P=0.000)。结论酶加去污剂法可较完全去除猪主动脉瓣膜细胞并保持细胞外基质的三维结构,但其生物力学性能有所下降;胎牛血清预处理能改善脱细胞瓣天然支架的细胞黏附、生长和繁殖。Objective To observe the mechanical properties of decellularized porcine aortic valve, and to explore the effects of precoating methods of biological scaffold on histocompatibihty. Methods Fresh porcine aortic valves were decellularized using trypsin, TritonX-100 and nuclease. Treated valves were evaluated by light microscopy, scanning electron microscopy （SEM） and mechanical test. Three groups of scaffold were precoated with phosphate buffered saline （ PBS）, poly-L-lysine （PLL） and fetal bovine serum （FBS） respectively. Myofibroblasts were seeded onto each scaffold. Light and electron microscopic observation was performed and MTT test was used to examine efficiency of cell attachment. Results HE stain and SEM showed that cells were almost absent in the treated leaflet. The wave-like collagen together with the whole three-dimensional structure was maintained. Compared with normal valves, the Max-load, Max-stress and elastic modulus decreased while the Max-strain increased （ P 〈 0.05 ）. The result of MTT test showed more cells were attached on the valves treated with FBS compared to the other two groups. Histological investigations also confirm that the high degree of cell attachment on the valves precoated with FBS（F = 129.26, P = 0. 000）. Conclusions Enzyme combined with detergent and nucleases can remove cells from porcine aortic valves. Meanwhile the mechanical properties of these valves may be altered. Precoating porcine aortic valve with FBS is an effective method to improve cell attachment, growth and increasing.

关 键 词：主动脉瓣 猪 生物力学 组织相容性 心脏瓣膜 

分 类 号：R318.08[医药卫生—生物医学工程]