单克隆抗体CD44-生物素-亲和素技术构建组织工程软骨修复猪膝关节负重区软骨缺损  

Construction of tissue engineering cartilage for the damaged load bearing section of pig knee-joint by CD44 monoclonal antibody-biotin-avidin binding technique

作　　者：林红[1] 周健[1] 王会仁[1] 沈龙祥[2] 郭常安[1] 董健[1] 陈峥嵘[1] 

机构地区：[1]复旦大学附属中山医院骨科,上海200032 [2]上海交通大学附属第六人民医院骨科

出　　处：《中华实验外科杂志》2014年第3期607-610,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30872623、81071468、81301577）;上海市科委基础重点科研项目（09JC1403600）;高等学校博士学科点专项科研基金资助项目（20130071120062）

摘　　要：目的 观察单克隆抗体CD44-生物素（Biotin）-亲和素（Avidin）绑定（CBA）技术,在体外构建的组织工程软骨移植修复猪膝关节负重区软骨缺损的效果.方法 分3组构建组织工程软骨：常规组：壳聚糖三维支架＋软骨细胞;Biotin-Avidin绑定（BA）技术组：生物素＋亲和素化壳聚糖三维支架＋软骨细胞;CBA技术组：生物素化单抗CD44＋亲和素化壳聚糖三维支架＋软骨细胞.分4组修复猪膝关节负重区软骨全层缺损：A组植入常规组织工程软骨;B组植入BA技术构建的组织工程软骨;C组植入CBA技术构建的组织工程软骨;D组植入自体软骨.12周后取标本行大体观察并行Moran评分;生物力学方法评估标本弹性模量和刚度;Western blot检测新生软骨组织内的Ⅱ型胶原（colⅡ）、聚集蛋白聚糖（aggrecan）目的蛋白含量;对标本行苏木素-伊红染色、甲苯胺蓝染色并行Wakitani评分.结果 Moran大体评分,D组为（4.48 ±0.23）分,与C组差异无统计学意义（P＞0.05）,但优于A、B两组（P＜0.05）;D组弹性模量为（20.61 ±±2.02） mPa,刚度为（72.28±6.16） N/mm、colⅡ及aggreean目的蛋白含量比值分别为1.59 ±0.14和1.44±0.13,均优于其他组（P＜0.05）;组织学染色及Wakitani评分均显示D组修复效果较好.结论 BA绑定技术比常规接种方法所构建组织工程软骨对猪膝关节负重区软骨缺损修复效果好,但是比较而言,单克隆抗体CBA绑定技术所构建组织工程软骨修复效果更佳,虽仍未达到自体移植修复水平.Objective To evaluate the effect of repairing cartilage defect using tissue engineering cartilage with CD44 monoclonal antibody-biotin-avidin （CBA） binding technique in vitro and in vivo.Methods The tissue engineering cartilages were designed and divided into 3 groups：general group,chitosan scaffold ＋ chondrocyte group; biotin-avidin （BA） group,chitosan scaffold bound with BA ＋ chondrocyte; CBA group,chitosan scaffold bound with CD44 monoclonal antibody-BA ＋ chondrocyte.The levelthrough cartilage damages of load bearing sections were divided into four groups：group A,transplanted with the tissue engineering cartilage in the general group; group B,transplanted with the tissue engineering cartilage in the BA group; group C,transplanted with the tissue engineering cartilage in the CBA group;group D,transplanted with the autologous cartilage.Samples were taken after transplantation for 12 weeks,and estimated by Moran estimate system.The modulus of elasticity and rigidity of the samples were estimated by biomechanics method.The contents of collagen Ⅱ （col Ⅱ） and aggrecan were tested by Western blotting.The samples were stained with hematoxylin and eosin （HE）,and TB dye,and the Wakitani scores were recorded.Results In the Moran estimate system,the score in group D was 4.48 ± 0.23,which showed no statistically significant difference from group C （P ＞ 0.05）,but higher than in groups A and B,P ＜ 0.05.The modulus of elasticity in group D was （20.61 ± 2.02） mPa and the rigidity was （72.28 ±6.16） N/mm.The expression of col Ⅱ and aggrecan in group D was 1.59 ±0.14 and 1.44 ± 0.13 respectively,significantly higher than other groups,P ＜ 0.05.The histologic staining and Wakitani score revealed the more satisfactory repair effectiveness in group D.Conclusion The repair effect of tissue engineering cartilage constructed by the BA technique for the damaged load bearing section of pig knee-joint is more satisfactory than the general method.The effect,however,is much better when t

关 键 词：单克隆抗体CD44 生物素 亲和素 组织工程 软骨缺损 

分 类 号：R392.1[医药卫生—免疫学]