血流力学对组织工程心脏瓣膜种子细胞形态及功能的影响

Effect of blood stream stress on the morphology and function of seed cells on tissue engineering heart valve

目的观察血流力学对组织工程心脏瓣膜种子细胞形态及功能的影响。方法从犬骨髓中分离得到骨髓间充质细胞(MSCs),经DiI标记后接种于去细胞猪主动脉瓣膜支架,培养7 d后将所得瓣叶种植于犬腹主动脉内,术后4、8、12周取材,进行HE染色、天狼猩红染色、免疫组织化学染色及超微电镜检查等,观察细胞生长、形态及功能变化。设体外培养组为对照组。结果接种骨髓间充质细胞的去细胞猪主动脉瓣膜支架,移植于犬腹主动脉内后,仍可检测到所种植细胞,细胞生长良好。与对照组比较,细胞形态有所伸长,整体上沿流体方向改变形态,同时,瓣叶组织中Ⅰ、Ⅲ型胶原含量明显增加,并充满中性和酸性黏多糖。结论构建组织工程心脏瓣膜时,血流力学对种子细胞维持形态、保持功能起着重要作用。

Objective To study the effect of blood stream stress on the morphology and function of seed cells on tissue engineering heart valve. Methods Isolated from dog bone marrow, MSCs were seeded on the decellularized porcine aortic valve bracket and cultured in cell culture medium after DiI labeling. The harvested valve cusp was implanted in the abdominal aorta after culture for 7 days. The samples were drawn on the postoperative 4,8 and 12 weeks respectively. TEHV was subjected to HE staining, immunohistochemical staining and electron microscopic examination to observe the changes of cell growth, morphology and function. The in vitro cultures served as control group. Results After implanted in the abdominal aorta, the decellularized porcine aortic valve bracket which was inoculated with the MSCs could be detected with the existence of implanted cells which developed well. Comparing with the control group, the cells were stretched along the fluid direction as a whole. Meanwhile, the contents of collagen type Ⅰand type Ⅲ in valve cusp were increased significantly. Conclusion Blood stream stress has a significant effect on maintaining the morphology and function of the seed cells when constructing the tissue engineering heart valve.