关节软骨源性多孔支架复合自体软骨细胞修复兔关节软骨缺损的长期疗效观察  

Long-term effects of articular cartilage derived scaffold/autologous chondrocytes composites for repairing full-thickness defects of knee articular cartilage in rabbits

作　　者：赵斌[1] 王玉[1] 黄靖香[1] 眭翔[1] 张莉[1] 田明[1] 王鑫[1] 汪爱媛[1] 许文静[1] 卢世璧[1] 

机构地区：[1]解放军总医院全军骨科研究所,北京市100853

出　　处：《中国矫形外科杂志》2010年第11期937-940,共4页Orthopedic Journal of China

基　　金：军队十一五计划(编号:06Z057)资助项目;国家自然基金重点项目(编号:30930092);国家支撑计划(编号:2006BAI16B04)资助项目

摘　　要：[目的]对关节软骨源性多孔支架复合自体软骨细胞复合体修复兔膝关节软骨缺损的长期效果进行观察和评价。[方法]实验动物新西兰大白兔共10只,分两组:(1)关节软骨源性支架对照组:缺损内置入关节软骨源性支架;(2)关节软骨源性支架复合细胞组:缺损内置入关节软骨源性支架-自体软骨细胞复合体。手术后15个月取材做大体摄像,甲醛固定、10%EDTA脱钙,石蜡切片,采用四种组织化学染色法(HE、奥新兰(AB)、甲苯胺兰(TO)、藩红花"O"染色)和Ⅱ型胶原免疫组化染色,观察修复后的兔关节软骨组织细胞形态特征。[结果]关节软骨源性支架对照组,大体观察关节表面有凹陷。石蜡切片HE染色见未修复缺损处为梭形细胞,纤维软骨,缺损处AB、甲苯胺兰、藩红花"O"染色为阴性,Ⅱ型胶原染色为阳性;关节软骨源性支架复合细胞组,大体见关节软骨表面修复平整、光滑。常规HE染色镜下见软骨全层修复良好,为透明软骨细胞,可见软骨陷窝潮线排列结构。细胞外基质特染:AB、甲苯胺兰、藩红花"O",Ⅱ型胶原特种染色均为阳性。[结论]组织学观察结果发现,关节软骨源性支架组为纤维软骨组织结构,未能完全修复膝关节软骨缺损;而关节软骨源性支架复合细胞组关节软骨缺损修复良好,未见退变,具有软骨组织的特征。[Objective]To observe the histological characteristics of autologous chondrocytes/human articular cartilage derived scaffold （HACDS） composites for repairing chondral defects in rabbit models. [Methods]A full-thickness articular-cartilage defect （diameter 4 mm,thickness 2 mm） was created in the femoral condyle of rabbits. The rabbits were divided into two groups： control group,HACDS scaffold only,and experimental group,chondrocytes/HACDS scaffold. Fifteen months after implantation,the specimens were observed by inverted phase contrast microscopy,and assessed by staining with haematoxylin-eosin,alcian blue,toluidine blue,safranin O,as well as by the immunohistochemistry of collagen type Ⅱ.[Results]Histologically,the generated neo-cartilage integrated well with its surrounding normal cartilage and subchondral bone in the defects of experimental group at 15 months post-implantation,whereas only fibrous tissue was filled in the defects of control group. Histochemical and immunohistochemical analysis revealed that alcian blue,toluidine blue,safranin,and were obviously positive in the experimental groups. However,alcian blue,toluidine blue,and safranino O were negative in the control group.[Conclusion]The current histological examination demonstrated that an engineered cartilage composed of autologous chondrocytes on HACDS scaffold could be successfully obtained and further applied to repair an articular cartilage defect in a rabbit model.

关 键 词：关节软骨源性支架 软骨细胞 关节软骨缺损 组织学评价 

分 类 号：R687.4[医药卫生—骨科学]