动物源性骨软骨支架修复兔膝关节复合缺损  

Repair of composite defect in rabbit knee joint using animal-derived osteochondral scaffold

作　　者：谭文成 林宏生[2,3] 张嘉晴[4] 郑立恒[5] 梁耀中[6] 夏吉生[5] 黄馨霈 吴昊[2,3] 陈舒[7] 查振刚[2,3] 

机构地区：[1]澳门医学科技研究协会,999078 [2]暨南大学附属第一医院 [3]暨南大学骨科疾病研究所,广州510630 [4]暨南大学医学院,广州510632 [5]澳门仁伯爵医院,999087 [6]暨南大学附属第一医院急诊科,广州510630 [7]南方医科大学附属花都人民医院,广州510800

出　　处：《中国骨科临床与基础研究杂志》2011年第1期51-59,共9页Chinese Orthopaedic Journal of Clinical and Basic Research

基　　金：澳门医学科技研究基金项目(031/2009/A);广州市科技计划(2008Z1-E411)

摘　　要：目的探讨软骨细胞-动物源性骨软骨支架复合体修复兔膝关节骨软骨复合缺损的可行性和影响因素。方法将改良贴壁离心法获取的骨髓间充质干细胞(bone marrowm esenchymal stem cells,BMSCs)/诱导分化的软骨细胞共培养后与经深低温冷冻、脱脂、脱钙、真空冷冻干燥和辐照消毒的动物源性骨软骨支架复合,构建共培养细胞+软骨-骨一体化复合支架。27只新西兰大白兔随机分为实验组(A组)、对照组(B组)和空白组(C组),每组9只。于兔股骨髁间窝处钻一深6mm的骨软骨复合缺损,A组植入共培养细胞+骨软骨复合支架,B组植入骨软骨复合支架,C组不植入任何支架材料和细胞,分别于术后4周、8周和12周取材,行大体观察、苏木精—伊红染色和甲苯胺蓝染色,并对各标本的软骨切片进行组织学评分。结果随着时间的延长,A组大体观察见复合缺损区已完全修复,局部无凹陷,新生组织和周围组织融合;B组新生组织仍不能完全填充缺损;C组缺损区仍明显。苏木精—伊红染色和甲苯胺蓝染色见A组软骨缺损区由新生的透明软骨样组织修复,细胞呈柱状排列,极性好,软骨陷窝明显,骨缺损区由骨样组织修复,新生软骨和软骨下骨以及宿主骨界面耦合良好;B组新生软骨细胞无软骨陷窝,排列混乱,各界面藕合欠理想;C组可见陈旧性肉芽组织生长并突出于缺损区表面。甲苯胺蓝染色阳性率和组织学评分结果表明,A组与B、C两组之间的差异具有统计学意义(P<0.05)。结论 BMSCs/诱导分化的软骨细胞共培养细胞复合动物源性骨软骨支架对兔膝关节软骨和软骨下骨的复合缺损具有修复作用。Objective To investigate the feasibility of chondrocytes-animal derived osteochondral scaffold composite for repairing osteochondral composite defects in rabbit knee.Methods The co-cultured cells,consisted of adherence separated bone marrow mesenchymal stem cells（BMSCs） /induced and differentiated chondrocytes,were cultured on the animal osteochondral scaffold which was processed with deep frozen,degreased,decalcified,vacuum freezely dried and radiation sterilized procedures to construct the co-cultured cellular osteochondral scaffold composite.Twenty-seven newzealand rabbits were randomly divided into experimental group（group A）,control group（group B） and blank group（group C）,and there were 9 rabbits in every group.After a 6 mm-depth osteochondral composite defect was made by drilling in the intercondylar fossa of femur,co-cultured cellular osteochondral scaffold composite were planted in group A,only osteochondral scaffolds were planted in group B,while neither scaffolds nor cellular composite were planted in group C.At 4,8,and 12 weeks postoperatively,the restoration for osteochondral composite defects were evaluated by gross observation,hematoxylin-eosin staining,toluidine blue staining and histological score.Results With time go on,gross observation showed that the defects in group A were recovered completely,the regeneration and surrounding tissues were integrated and there was no hollow,while the new connective tissue in group B could not completely cover the defects,and the defects in group C were not recovered at all.Hematoxylin-eosin and toluidine blue staining revealed that there were a lot of new hyaline cartilage at cartilage defect area in group A.In the new growth tissue columnar cells and cartilage lacuna were well arranged and polarized,also,there were many osteoid tissues in the bone defects,the interface of the regenerative cartilage,the subchondral bone and host bone coupled well.In group B,there were no cartilage lacuna in the regeneration cartilage,the arrangement of the cell

关 键 词：软骨细胞 骨髓细胞 干细胞 细胞 培养的 支架 膝损伤 软骨 关节 兔 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R684[医药卫生—基础医学]