南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (01): 22-25.

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胰蛋白酶法和Triton-X100法去除瓣膜细胞的效果比较

王克学, 张镜方, 詹秋鹏, 简旭华   

  1. 广东省心血管病研究所心脏外科, 广东广州510080
  • 出版日期:2005-01-20 发布日期:2005-01-20
  • 基金资助:
    收稿日期:2004-7-25。
    基金项目:广东省自然科学基金重点项目(C-010177)
    作者简介:王克学(1972- ),男,广东省心血管病研究所在读博士研究生,主治医师,电话:020-83827812-41403,E-mail:kexuewang@tom.com

Effect of trypsin and Triton-X 100 for decellularization of porcine aortic heart valves

WANG Ke-xue, ZHANG Jing-fang, ZHAN Qiu-peng, JIAN Xu-hua   

  1. 广东省心血管病研究所心脏外科, 广东广州510080
  • Online:2005-01-20 Published:2005-01-20

摘要: 目的 比较胰蛋白酶法和Triton-X100法去除异种或同种心脏瓣膜上细胞的效果。方法 选取新鲜猪主动脉瓣膜片分别置入0.05%胰酶溶液和0.25%Triton-X100溶液中处理48h,并以未经处理的新鲜瓣膜片作对照,行光镜及电镜观察,并作热皱缩温度、断裂强度和断裂拉伸率测定。结果 光镜和电镜下均显示,胰酶法和Triton-X100法均彻底去除了主动脉瓣膜上的细胞成分,但Triton-X100法对瓣膜支架结构的损伤较大。热皱缩温度、断裂强度和断裂拉伸率在新鲜瓣膜组和胰酶组间无显著性差异,但Triton-X100组与新鲜瓣膜组及胰酶组相比均有下降。结论 胰酶法去除猪主动脉瓣膜细胞彻底,保持了瓣叶支架结构,优于Triton-X100法。

Abstract: Objective To test two decellularization procedures for their potential of cell removal and mormal matrix preservation. Methods Porcine aortic valve leaflets were treated with either 0.05% trypsin or 0.25% Triton-X 100 respertively for 48 h for decellularization and with fresh untreated valve leaflets as control. Two tissue samples from each group were stained with hematoxylin and eosin and observed light-microscopically followed by scanning electron microscopy. Ten valve leaflets in each group were measured for shrinkage temperature, tensile strength/fracture toughnes and percentage elongation. Results Trypsin and Triton-X 100 all achieved complete decellularization but Triton-X 100 caused stronger structural alterations. No significant difference was identified between untreated and trypsin groups in shrinkage temperature, tensile strength/fracture toughnes and percentage elongation, but Triton-X 100 group showed significant difference from the other two groups. Conclusion Decellularization using trypsin is superior to Triton-X100 in efficiency and matrix preservation.

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