组织工程血管管状支架的制备与动态培养在血管组织工程中的应用

Preparation of tubular scaffold for tissue engineering vessel and the application of dynamic implantation and culture in vascular tissue engineering

目的探索体外构建组织工程血管及细胞种植的方法。方法（1）制备管状支架模具，采用模具成形，冷冻干燥等技术，应用胶原和透明质酸复合体外预构管状支架；（2）分动态种植和静态培养两组进行细胞分层种植，每组12个，应用水溶性磺酸四唑（WST-1）测定，扫描电镜观察，5-溴2-脱氧嘧啶核苷（BrdU）细胞标记鉴定比较两种方法的细胞生长情况。结果（1）成功制备了管状支架，在培养液中约3周可保持管腔形态不变形，不塌陷；2＋WST-1测定值在培养6、14、20d分别为：动态组0．842±0．110、1．124±0．102、1．132±0．112，静态组0．386±0．121、0.489±0．245、0．499±0．265，两组比较差异有统计学意义，电镜和BrdU标记表明动态组细胞排列整齐有序。结论应用模具成形，冷冻干燥技术预构管状支架的方法切实可行，动态培养有助于细胞种植，本研究为组织工程血管的构建方法提供了思路。

Objective To study the method of tubular scaffold and the application of dynamic implantation and culture in vascular tissue engineering. Methods （1） A special model for tubular scaffold was made and tubular scaffold was constructed with collagen and sodium hyaluronic acid using the techniques of vacuum formation and freeze-drying. （2） Vascular endothelial cells and smooth muscle cells of rabbits were cultured, and implanted in the tubular scaffolds sequentially. Stated or dynamic implantation and culture method were compared using WST-1 method and SEM. Results The tubular scaffold was made successfully with elasticity, stubbornness and hydrophilic and could maintain their shapes in culture medium for long time. wst-1 levels in the cells cultured for 6, 14 and 20 days were 0. 842 ± 0. 110, 1. 124 ± 0. 102,1. 132 ± 0.112 in rotary cell culture system, and 0.386 ± 0. 121,0. 489 ± 0.245,0. 499 ± 0. 265 in static culture system respectively. Cells grew faster in dynamic group than in stated group. Cells arranged regularly in dynamic group while irregularly in static group. Conclusion （ 1 ） The techniques of model-formation and freeze-drying to construct the tubular scaffold are feasible; （2） Dynamic implantation and culture help to improve the cell adhesion and proliferation.