TritonX-100法制备牛颈静脉脱细胞血管支架

Bovine Jugular Vein Decellularized with Triton X-100 as a Potential Scaffold for Vascular Tissue Engineering

【目的】研究经TritonX-100脱细胞处理的牛颈静脉的组织结构和生物力学特性以及作为组织工程血管支架的可能性。【方法】获取10条新鲜带瓣牛颈静脉,随机分为两组,新鲜对照组(n=5)和脱细胞实验组(n=5),脱细胞组血管使用2.5mL/LTritonX-100等进行脱细胞处理48h,病理切片染色和扫描电镜观察血管壁和瓣膜自身细胞的脱除情况和细胞外基质变化情况;生物力学性能检测血管组织强度变化;体外人内皮种子细胞种植以了解脱细胞支架的生物相容性。【结果】脱细胞处理后,管壁和瓣膜的自身细胞完全脱除而弹力纤维和胶原纤维无明显改变;与新鲜牛颈静脉相比,脱细胞血管的拉伸强度[(16.5±2.61)MPavs(15.5±3.1)MPa;P>0.05]和最大持线力[(7.9±0.9)Nvs(7.0±1.1)N;P>0.05]无明显改变,在100mmHg液压下脱细胞血管无异常扩张;内皮种子细胞在脱细胞血管支架上生长黏附良好。【结论】TritonX-100法对新鲜牛颈静脉进行脱细胞处理效果良好,脱细胞牛颈静脉可作为细胞种植的血管支架使用。

[Objective]To study the composition and strength of bovine jugular vein decellularized with Triton X-100 and to determine its potential as a vascular tissue-engineering scaffold. [ Methods] Fresh bovine jugular veins （n= 10）were obtained and divided into two groups （n =5） at random, the fresh group and the decellularized group, specimens in decellularized group were treated with 2.5 mL/L Triton X-100 and sodium-deoxycholate for 48 h. Residual cells and extracellular matrix composition were studied with light and electron microscopy. The strength of graft was measured in vitro by insufflation and pull-through techniques. Decellularized specimens were seeded with human endothelial cells and cultured for 7 d. [Results] 2.5 mL/L Triton X-100 together with sodiumdeoxycholate completely removed native bovine cells. Collagen morphology and elastin staining appeared unchanged. Compared with fresh bovine jugular vein, decellularized vein had similar in vitro tensile stress [ （16.5 ± 2.6）MPa vs （15.5 ± 3.1）MPa; P 〉 0.05] and suture-holding strength [（7.9 ± 0.9）N vs （7.0 ± 1.1）N; P 〉 0.05], and had no abnormal dilation under 100 mm Hg intraluminal pressure. Endothelial cells attached to acellular specimens and grew well. [Conclusion] Bovine jugular vein rendered acellular with Triton X-100 presented an excellent scaffold for recellularization with human cells.