外源性软骨细胞在组织工程化软骨组织形成中的作用  

Effects of exogenous chondrocytes in the formation of tissue-engineered cartilage tissues

作　　者：孙安科[1] 裴国献[1] 胡平[2] 陈江睿[2] 任高宏[1] 胡罢生[1] 刘晓静[1] 张宇[1] 覃昱[1] 

机构地区：[1]解放军第一军医大学南方医院全军创伤骨科中心组织工程研究中心,广东省广州市510515 [2]清华大学化学工程系高分子研究所,北京市100084

出　　处：《中国临床康复》2004年第35期7948-7949,7969,i001,共4页Chinese Journal of Clinical Rehabilitation

基　　金：国家重点基础研究发展规划(九七三):组织工程的基本科学问题(G1999054309);中国博士后科学基金资助金资助(中博基犤2002犦11号文件)~~

摘　　要：目的:观察腺病毒介导的绿色荧光蛋白(greenfluorescentprotein,GFP)标记软骨组织工程种子细胞的可行性,探讨外源性软骨细胞在组织工程化软骨组织形成中的作用。方法:实验于2002-01/08在解放军第一军医大学实验室完成。腺病毒GFP表达载体以巨细胞病毒(CMV)为启动子转染体外培养软骨细胞作为种子细胞与poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)(PHBHH)复合后植入兔体内,术后一定时间取材,确定组织工程化组织的表型,观察GFP的表达。结果:以CMV为启动子的腺病毒GFP载体能有效转染体外培养软骨细胞,转染效率达53.66%。携带GFP的软骨细胞作为软骨组织工程种子细胞,6周和12周形成的组织工程化组织中检测到GFP表达,经苏木精-伊红染色、免疫组织化学检查和特殊染色确定获取的组织为软骨组织,且与未转染软骨细胞形成的组织工程化软骨组织形态学一致。结论:腺病毒介导的GFP能有效转染体外培养软骨细胞,外源性软骨细胞是体内组织工程化软骨组织形成的源细胞。AIM:To investigate the feasibility of tissue-engineered chondrocytes as seed cells labeled by adenovirus-mediated green fluorescent protein(GFP) and probe into the effects of exogenous chondrocytes in the formation of tissue-engineered cartilage tissues. METHODS:The experiment was completed in the Laboratory of First Military Medical University of Chinese PLA from January to August 2002.After the chondrocytes cultured in vitro were transfected by expression vector of adenovirus-mediated GFP gene with cytomegalovirus(CMV) as the promotor,they were used as seed cells to combine with poly (3-hydroxybutyrate-co-3- hydroxyhexanoate)(PHBHH) scaffold to form cells-PHBHH complexes,which were then implanted into subcutaneous tissue of rabbits.The chondrogenesis and GFP expression were evaluated in different period after implantation. RESULTS:The expression vector of adenovirus-mediated GFP gene with CMV as the promotor could efficiently transfect the chondrocytes cultured in vitro,and the transfection rate was 53.66%.The GFP chondrocytes with GFP gene could be used as the seed cells of cartilage tissue engineering.GFP expression was respectively found in the tissue-engineered tissues 6 and 12 weeks after implantation and these tissue-engineered tissues were determined as cartilage tissues by hematoxylin-eosin(HE) staining,immunohistochemical examination and some special staining.Furthermore,the neocartilage was morphologically the same as the tissue-engineered cartilages formed by chondrocytes unlabelled with GFP. CONCLUSION:Adenovirus-mediated GFP can effectively transfect the chondrocyte cultured in vitro.The tissue-engineered cartilage tissues in vivo originate from exogenous chondrocytes.

关 键 词：生物医学工程 腺病毒科 软骨 软骨细胞 发光蛋白质类 

分 类 号：R318[医药卫生—生物医学工程]