组织工程化血管构建的初步实验研究

Primary experimental study on the construction of tissue engineering blood vessel

目的探讨组织工程化血管构建的初步实验方法。方法预构具有三维结构的胶原蛋白几丁聚糖骨架,以人血管内皮细胞、平滑肌细胞及成纤维细胞作为种子细胞,培育后二步法种植并行工程化血管成熟培养。经生物学检测,补片修补实验鼠腹主动脉人为缺损。结果人血管内皮细胞、平滑肌细胞及成纤维细胞可作为种子细胞应用;预制的三维骨架具有良好的组织和细胞兼容性,种植种子细胞后可形成良好的细胞黏附、生长;体外静态及生物反应器培养,工程组织中细胞数及细胞外基质含量明显增加(P<0.05);血小板凝集试验表明该工程组织具有一定的抗凝特性;修补大鼠腹主动脉缺损,术后10d可维持通畅。结论胶原蛋白几丁聚糖预构的三维骨架可应用于血管组织工程研究,细胞种植后经成熟培养,可初步构建具有一定强度和抗凝特性的组织工程化血管。

Objective To explore the primary experimental methods to construct tissue engineering blood vessel. Methods Using the collagen-chitosan to prefabricate the scaffolds with 3-dimentional structure, the proliferated human endothelial cells （ECs）, smooth muscle cells （SMCs） and fibroblasts act as the seed cells. The cells were seeded to scaffolds in two-step method, and engineering tissue were matured by static culture or bioreactor culture. Extracellular matrix contents and the platelet aggregation were examined in engineering tissue, tissue engineering blood vessels were taken as patches to repair the manmade defaults on the rats aorta. Results The proliferated human ECs, SMCs and fibroblast can hold activity and act as seed cells. The prefabricated scaffolds, with excellently cell and tissue biocompatibility, can facilitated cells adherence and upgrowth, the cells quantities and extracellular matrix contents in engineered tissue are time dependent increase （ P 〈 0. 05 ）. Platelet aggregation tests confirm the tissue engineering blood vessel have some anti-coagulability. Using the engineering tissue patch to repair the default, 6 aortas in 8 animal were patency till 10 days post-operation. Conclusions The seeding cells can be seeded on the 3-dimensional collagen-chitosan scaffolds and matured, the tissue engineering blood vessel can be constructed primarily.