脐动脉在搏动流作用下脱细胞制备小口径组织工程血管支架的研究

Pulsatile Flow Used for Small Diameter Blood Vessel Scaffold Decellularized by Human Umbilical Artery

探索利用搏动流对小口径血管进行脱细胞的方法。首先将脐动脉连接到搏动流管道中进行灌注,结合0.25%胰蛋白酶和0.01%乙二胺四乙酸混合溶液的共同作用,以1%十二烷基硫酸钠为脱细胞试剂对脐动脉洗脱3h。洗脱3h后的病理切片显示脐动脉细胞被全部脱去;拉伸实验测定脐动脉脱细胞前极限应力为(3.55±0.42)N,脱细胞后其极限应力为(3.50±0.43)N(P>0.05);在300mmHg压强作用下,脐动脉脱细胞前后两组(各30根血管)均有2根破裂,28根保持完好(P>0.05)。此结果预示,脐动脉在脱细胞前后的力学特性无显著差异。成纤维细胞静态培养显示,成纤维细胞能在脱细胞脐动脉支架表面良好生长。研究结果表明利用搏动流将小口径血管脱细胞是一种简便、快速、有效的方法。

To explore the method of using pulsatile flow to decellularize small diameter blood vessels. First , the human umbilical arteries were poured with detergents and treated with 0.25% trypsin/0.01% EDTA 30 minutes. Then they were connected into a pulsatile flow circuit and decellularized by 1% sodium dodeeyl sulfate （SDS） for 3 hours, when the arteries were completely aeellularized and observed by pathologic examination. The tests on mechanics to the arteries indicated that the maximum tensile stress of the arteries was （ 3.55± 0.62 ） N before decellularization and （ 3.50 ± 0.43 ） N after decellularization（ P 〉 0.05 ）. Two vessels ruptured and 28 vessels remained intact in the pressure range of 300 mmHg before and after decellularization. This test indicated that there was no significant differenee in mechanical properties between treated arteries and untreated arteries. Additionally the static cultivation of fibroblasts displayed that the fihrohlasts grew well on the umbilical artery seaffolds sheet. These results suggest that small diameter blood vessels can be decellularizied in pulsatile flow condition conveniently, rapidly and efficiently.