自体骨髓间充质干细胞与外源性透明质酸钠修复兔膝关节软骨缺损的研究

目的探讨全层软骨缺损修复效果与关节腔内骨髓间充质干细胞(marrow mesenchymal stem cells,MSCs)来源数量的相关性,了解体内条件下外源性透明质酸钠(sodium hyaluronate,SH)能否促进体外培养扩增的MSCs聚集到软骨缺损处. 方法 4月龄日本大耳白兔66只,用手摇钻制成直径5 mm、深4 mm的圆柱形膝关节软骨缺损.从兔股骨提取MSCs, 经体外培养Brdu标记后,单独或与SH一起注射到自体软骨损伤关节腔中.实验动物共分4组:A组(n=15),同时注射SH和兔自体MSCs;B组(n=15),单独注射自体MSCs;C组(n=18),单独注射SH;D组(n=18),不给治疗措施.术后5、8和12周,行大体、组织学、免疫组织化学观察及修复组织厚度测定. 结果术后5、8和12周,修复组织厚度A组与B组比较,差异有统计学意义(P＜0.01);A组与C组比较及B组与D组比较,差异无统计学意义(P＞0.05).组织学观察显示,修复组织A组以纤维软骨为主,B组以纤维组织为主.两组免疫组织化学染色缺损部位均未见到Brdu标记物. 结论关节腔内注入体外培养扩增的MSCs,不能促进软骨缺损修复.外源性SH对软骨缺损的修复有一定作用,但不能将MSCs聚集到缺损处和提高其趋化能力.

EFFECT OF AUTOLOGOUS BONE MARROW MESENCHYMAL STEM CELLS AND EXTROGENOUS SODIUM HYALURONATE ON REPAIRING KNEE JOINT DEFECT IN RABBITS

Objective To explore the relationship of the limited resource of the autologous bone marrow mesenchymal stem cells (MSCs) in articular cavity to the treatment results of full thickness articular cartilage defect, and to investigate whether the extrogenous sodium hyaluronate(SH) promotes the migration of MSCs cultured in vitro to the articular defect in vivo. Methods Sixty six Japan rabbits were made the model of the full thickness articular cartilage defect (5 mm width and 4 mm depth). The autologous MSCs were extracted from the rabbit femur, cultured in vitro, labeled by Brdu, and injected into the injured articular cavity with or without SH. The experiment was divided into 4 groups; group A (MSCs and SH, n =15); group B (MSCs, n =15); group C (SH, n =18); and group D (non treatment, n =18). The morphologic observation was made by HE staining, Mallory staining and immunohistochemical staining after 5 weeks, 8 weeks and 12 weeks of operation. Results There were significant differences in the thickness of repairing tissue between group A and group B( P <0.01); but there were no significant differences between group A and group C, and between group B and group D( P>0.05) . The histological observation showed that the main repairing tissue was fibrocartilage in group A and fiber tissue in group B. Conclusion MSCs cultured in vitro and injected into the articular cavity can not improve the treatment results of the articular cartilage defect. Extrogenous SH has effect on repairing cartilage defect. The extrogenous SH has no effect on the chemotaxis of the MSCs, and on the collection of MSCs into the joint defect.