超高压脱细胞组织工程血管支架的实验研究

Experimental Study on Acellular Heart Valves and Vessels by Ultrahigh Pressure Treatment

目的采用新的更安全的抗原去除技术-超高压脱细胞方法处理同种异体血管,观察该方法的脱细胞效果以及猪内源性逆转录病毒(PERV)的去除情况。方法无菌条件下取出猪降主动脉血管,用10 000atm超高静水压(4℃冷方压)将供体来源细胞压碎,结合酶的消化作用,PBS的搅拌洗涤,脱去细胞残片形成血管生物支架。对该支架脱细胞效果进行组织学评估、超微结构观察、检测DNA含量同时检测PERV。结果组织学显示超高压脱细胞方法可完全除去血管支架内的供体细胞成分,经超高压脱细胞处理后DNA含量显著降低(p<0．0001)、PERV被成功灭活。结论超高压脱细胞方法可为我们提供更安全可靠的组织工程血管支架。

Objective In this study, our novel tissue processing of decellularization by ultrahigh pressure treatment for safe tissue transplantation is reported. Method Porcine vessels were isolated under sterile conditions and treated by cold isostatic pressing (CIP) at 4℃for disruption of donor cells. The cell debris was then washed out in PBS and enzyme. Result The tissues were completely cell free when they were treated by a CIP of 980 MPa (10 000 atm) for 10 min. There was no porcine endogeneous retrovirus (PERV) detected in the treated tissue. The tissues were disinfected when CIP was applied to the tissues contaminated by normal bacteria floras. The content of DNA was significant degraded in the treated tissue. Conclusion This novel tissue processing of decellularization by ultrahigh pressure treatment for safe tissue transplantation may provide a more durable and safe bioscaffold for tissue transplantation.