注射性C/GP复合自体MSCs修复兔关节软骨的实验研究

Experimental Study on the Repair of Articular Cartilage by the Injectable C/GP Encapsulating Autoallergic MSCs

目的探讨可注射性壳聚糖/磷酸甘油(C/GP)复合自体骨髓间充质干细胞(MSCs)对兔关节软骨缺损修复的作用。方法将54只3个月新西兰大耳白兔双下肢髁关节面上制作2个直径3 mm、深3 mm的全层软骨缺损,随机分成A,B,C3组,每组18只。术后1周,A组注入壳聚糖/磷酸甘油(C/GP)复合自体MSCs混悬液1 mL;B组注入C/GP混合液1 mL;C组注入生理盐水1 mL,3组分别术后4、8、12周各处死大耳白兔6只,应用大体观察、组织学观察及组织学Wakitani评分评估各组大耳白兔缺损软骨的修复情况。结果大体观察:术后12周A组修复区与正常组织难以区别,B组修复区与正常组织接近但仍可区分;C组缺损明显,可见肉芽组织部分充填缺损。组织学观察:A组术后12周组织切片显示新生组织中见细胞体积较大,数量较多,与周围正常软骨组织相近;B组术后12周见短小卷曲的胶原纤维和软骨陷窝样组织;C组术后12周见纤维组织颠充部分缺损。组织学评分:A组4、8、12周组织学评分明显低于B、C组(P<0.01);B组4、12周组织学评分低于C组(P<0.01)。结论可注射性C/GP复合自体MSCs提高了对兔关节软骨缺损的修复效果。

Objective This study was designed to research the method of repairing articular cartilage defects in rabbits by the injectable chitosan /glycerophosphate（C/GP） Encapsulating autoallergic mesenchymal stem cells（MSCs）.Methods 54 3-months-old New Zealand rabbits were selected for making 2 models of articular cartilage defects in articular facets of both lower extremities,with 3 mm diameter and 3 mm depth.Randomly divided three groups A,B,C and every group was eighteen rabbits.After a week,A group injecte C/GP encapsulating autoallergic MSCs one ml,B group injecte C/GP one mL,C group injecte one ml normal sodium.Every 6 animals were sacrificed at 4,8,12 weeks postoperatively.The observation of macroscopy and the score of histology with Wakitani were used to evaluate the repairs of knee joint articular cartilage defects in rabbits.Results At 12 weeks postoperatively,A group in general observation difficult to difference with normal tissue;B group in general observation approximation with normal tissue;C group in general observation defection is obvious and it is thus clear that granulation tissue in defection.At 12weeks postoperatively,A group histological observation showed that there were many big cells volume and near to around normal cartilaginous tissue;B group histological observation showed that there were some microsoma capreolary collagen fibrils and cartilage lacuna;C group histological observation showed that fibrous tissue stuffed segmental defection.A group of the histological score in 4,8,12 weeks was lower than that in others groups（P〈0.01）;B group of the histological score in 4,12 weeks was lower C group（P〈0.01）.Conclusion The C/GP scaffold compounded with autoallergic MSCs transplantation can improve the repair effect on articular cartilage defects in rabbits.