去细胞猪主动脉瓣膜超微结构及生物相容性研究

Ultrastructure and biocompatibility of decellularized porcine heart valve

目的 观察曲拉通X-100（Triton X-100）与脱氧胆酸钠（SD）对猪主动脉瓣的去细胞效果,探讨去细胞猪主动脉瓣膜超微结构及其生物相容性.方法 分别采用1％ Triton X-100和1％SD对猪主动脉瓣进行去细胞处理,通过苏木素-伊红（HE）、Van Gieson染色,扫描电子显微镜从组织学上对去细胞瓣观察,通过血小板黏附实验和细胞毒性实验评价去细胞瓣的生物相容性.结果 1％SD去除细胞完全,细胞外基质保存完整,但去细胞瓣膜血小板黏附严重且具有一定的细胞毒性,24h和48 h的细胞相对增殖率分别为（80.29±2.05）％和（75.44±3.22）％;1％ Triton X-100去除细胞不完全,血小板黏附严重,也具有一定的细胞毒性,24h和48 h的细胞相对增殖率分别为（88.05±1.82）％和（82.95 ±3.07）％.结论 1％SD联合核酸酶对猪主动脉瓣的去细胞效果优于1％ Triton X-100,但均不够理想,其他无毒环保的去细胞试剂有待进一步探索.

Objective To evaluate ultrastructure and biocompatibility of decellularized porcine heart valve with Triton X-100 and sodium deoxycholate.Methods Porcine aortic valve leaflets were treated with 1% Triton X-100 and 1% sodium deoxycholate, respectively.The hematoxylin-eosin staining, Van Gieson staining and scanning electron microscopy were used to examine ultrastructure.Biocompatibility was evaluated by platelet adhesion and cytotoxicity test.Results 1% sodium deoxycholate allowed a thorough ceils removal with optimal matrix preservation, and the valves relative growth rate at 24 h and 48 h were （80.29 ±2.05）% and （75.44 ±3.22）%, respectively.1% Triton X-100 could not remove cells incompletely, and the valves relative growth rate at 24 h and 48 h was （88.05 ± 1.82）% and （82.95 ± 3.07） % respectively.These valves had a certain cell toxicity, and there were many platelets adhere to the valves.Conclusion The effect of decellularized heart valves with 1% sodium deoxycholate was superior to 1% Triton X-100, tut they were not ideal.Other avirulent environmental detergents need to explore in future.