摘要
目的探讨海藻酸钠支架复合组织工程化软骨细胞对兔佐剂性关节炎膝关节软骨缺损修复的影响。方法将新西兰白兔分为正常对照组、佐剂性模型组和海藻酸盐支架治疗组。模型组和治疗组兔膝关节内注入弗氏佐剂建立佐剂性关节炎模型。抽取治疗组兔左后健侧肢体的骨髓,用Percoll法分离出自体骨髓间充质干细胞(ABMSC),体外培养传代至P3代并诱导分化为软骨细胞。将软骨细胞与海藻酸钠支架混合注入治疗组的病变膝关节。对照组和模型组关节注射等量的生理盐水。海藻酸钠支架复合组织工程化软骨细胞治疗1个月后处死兔,应用Ⅱ型胶原免疫组织化学法染色和HE染色,观察各组兔的膝关节关节软骨缺损的修复情况。结果诱导成功软骨细胞且生长良好,呈现明显的纺锤体形,细胞成活率达98.5%。软骨细胞与海藻酸钠复合后体外培养显示细胞生长旺盛,增殖成株状或岛状,株状细胞团周围有类似的软骨陷窝形成,细胞排列密集,核圆形。组织形态学结果显示,治疗组与模型组比较,治疗组关节炎症反应明显减轻,关节Ⅱ型胶原阳性软骨细胞显著增高,软骨缺损得到明显修复。结论海藻酸钠支架复合组织工程化软骨细胞对兔佐剂性关节炎膝关节软骨缺损有一定的修复作用。
Objective To explore the effect of cartilage defect repair by using engineered chondrocyte composite with bone marrow stem cells （BMSCs） cultured on the sodium alginate scaffold in adjuvant-induced arthritis （AA） rabbits. Methods Twenty-four New Zealand white rabbits were divided into control group, model group and alginate treatment group at random. Complete Freund＇ s adjuvant was used to induce adjuvant- induced arthritis in model group and alginate treatment group. Bone marrow of the rabbits＇ healthy left hindlimbs was collected from the alginate treatment group, autologous bone marrow mesenchymal stem cells （ABMSCs） were isolated by the method of Percoll. The third generation （P3） ABMSCs were cultured and differentiated into chondrocytes. Engineered chondrocytes composite with sodium alginate scaffold were injected in the alginate treatment group, and the control group and model group were injected with saline in rabbits＇ articulus. Rabbits were sacrificed after one month of the treatment. Repair of articular cartilage defects were observed by HE and typeⅡ collagen immunohistochemical staining in rabbits＇ articular tissue.Results Engineered chondrocytes grew well and showing spindle-shaped, cell survival rate was 98.5%. Chondrocytes with sodium alginate composite also grew well in vitro, appearing plant-like or island, similar cartilage lacuna formed around the cells. The cells arranged densely, nucleus showing round. Compared with model group,histomorphometric results showed that inflammation was significantly reduced and type Ⅱ collagen positive cells remarkably increased in alginate treated rabbits. The repair of cartilage defect was significantly improved in alginate treatment group. Conclusions Engineered chondrocytes composite with alginate scaffold can repair cartilage defect in adjuvant-induced arthritis rabbits.
出处
《北京生物医学工程》
2013年第3期276-281,共6页
Beijing Biomedical Engineering
基金
浙江省科技计划项目(2009C33139)资助
关键词
佐剂性关节炎
海藻酸钠支架
自体骨髓间充质干细胞
软骨缺损
adjuvant-induced arthritis
sodium alginate scaffold
autologous bone marrow mesenchymal stem cell
cartilage defect
