同种异体和异种来源骨髓间充质干细胞诱导成软骨修复喉软骨缺损

Allogeneic versus heterogeneous bone marrow mesenchymal stem cells for laryngeal cartilage repair

背景:由于软骨细胞缺乏再生能力,选择合适的种子细胞是修复软骨缺损需首要解决的问题。目的:探讨同种异体和异种来源骨髓间充质干细胞成软骨诱导后修复喉软骨缺损的效果。方法:取第3代人骨髓间充质干细胞和兔骨髓间充质干细胞加入软骨定向诱导液(含转化生长因子β1和骨形态发生蛋白)进行成软骨诱导,并滴加于聚乳酸-羟基乙酸共聚物支架上。取30只新西兰大白兔随机分为3组:空白对照组、人骨髓间充质干细胞修复组和兔骨髓间充质干细胞修复组,制备喉软骨缺损动物模型,分别植入生理盐水浸湿的聚乳酸-羟基乙酸共聚物支架,人骨髓间充质干细胞复合聚乳酸-羟基乙酸共聚物支架,兔骨髓间充质干细胞复合聚乳酸-羟基乙酸共聚物支架。术后4周和8周,免疫组化法检测喉部组织Ⅱ型胶原的表达。结果与结论:各组动物均呼吸通畅、正常,未出现喘鸣等,进食和活动情况良好,未出现化脓或者感染现象。术后4周和8周,人骨髓间充质干细胞修复组和兔骨髓间充质干细胞修复组的Ⅱ型胶原阳性率均显著高于空白对照组(P<0.05)。人骨髓间充质干细胞修复组和兔骨髓间充质干细胞修复组间比较,差异无显著性意义(P>0.05)。结果表明同种异体和异种来源的骨髓间充质干细胞成软骨诱导后进行兔喉软骨缺损修复均可以获得良好的效果,二者修复效果无显著差异。

BACKGROUND： Because chondrocytes have no regeneration ability, to select suitable seed cells is the primary problem to repair cartilage defects. OBJECTIVE： To investigate the effect of allogeneic versus heterologous bone marrow mesenchymal stem cells（BMSCs） in repairing laryngeal cartilage defects after chondrogenic induction. METHODS： BMSCs from human and rabbits were isolated and cultured. Passage 3 cells were cultured in chondrogenic induction medium containing transforming transforming growth factor beta 1 and bone morphogenetic protein, and then were dropped onto a poly（lactic-co-glycolic acid）（PLGA） scaffold. Thirty New Zealand rabbits were randomly assigned into three groups： blank control group, human BMSCs group, rabbit BMSCs group. Animal models of laryngeal cartilage defects were made in the three groups. After modeling, saline-soaked PLGA scaffold, PLAG scaffold with human BMSCs or with rabbit BMSCs were implanted respectively into the rabbits in the normal blank, human BMSCs and rabbit BMSCs groups. The expression of type II collagen in the larynx and its surrounding tissues was detected by immunohistochemistry at 4 and 8 weeks postoperatively. RESULTS AND CONCLUSION： The animals in each group breathed normally with no presence of wheezing, and their eating and activity were good. Moreover, there was no purulency or infection in the three groups. At 4 and 8 weeks after operation, the positive rates of type II collagen in the two BMSCs groups were significantly higher than that in the blank control group（P 0.05）. There was no significant difference between two BMSCs groups（P 0.05）. These results show that both allogeneic and heterologous BMSCs have good therapeutic effects on the repair of laryngeal cartilage defects in rabbits.