应用可注射性生物材料再生自体组织工程化软骨组织的实验研究  

An experimental study of tissue engineered autologous cartilage by using an injectable polymer

作　　者：夏万尧[1,2,3] 曹谊林[1,2,3] 商庆新[1,2,3] 刘彦春[1,2,3] 刘德莉[1,2,3] 徐蓉[1,2,3] 吴娟娟[1,2,3] 

机构地区：[1]上海第二医科大学组织工程研究中心 [2]上海市组织工程研究中心重点实验室,上海200011 [3]上海第二医科大学附属第九人民医院整形外科,上海200011

出　　处：《中华整形外科杂志》2001年第5期302-305,共4页Chinese Journal of Plastic Surgery

基　　金：国家自然科学基金 ( 39840 0 0 6);上海市科学技术委员会基金 ( 9741190 0 8)资助

摘　　要：目的　探讨自体组织工程化软骨在动物自体内形成的最佳细胞浓度 ,为临床应用提供理论依据。方法　取贵州香猪部分耳廓软骨组织 ,于体外消化分离软骨细胞 ,与可注射性生物材料PluronicF12 7混合形成复合物 ,浓度为 10 ,2 0 ,3 0 ,4 0 ,5 0 ,60和 70× 10 6 ml,并将其接种于猪自体腹外侧壁皮下 ,于第 6周取材 ,分别进行大体、组织学、氨基糖氨多糖 (GAG)和Ⅱ型胶原含量检测 ,对再生软骨组织进行评价。结果　细胞浓度为 5 0× 10 6 ml时 ,再生的软骨组织 ,软骨细胞均匀分布 ,包埋在软骨陷窝内 ,与正常软骨组织有相似的组织学特征。而细胞浓度为 10× 10 6 ml、3 0× 10 6 ml则形成软骨组织不完全 ,软骨细胞散在分布 ,并被未降解的生物材料分隔。对所有样本称重 ,在 3 0～ 110mg之间 ,各样本GAG含量在 5 .8%～ 9.2 %之间 ,正常耳软骨含量为 9.2 %。WesternBlot证实所有样本都有Ⅱ型胶原。结论　利用组织工程技术可在具有免疫功能的自体动物内形成软骨组织 ,并确定了软骨组织形成的最佳细胞接种浓度为 5 0× 10 6 ml。Objective To investigate the proper cell density of tissue engineered autologous cartilage to indicate the clinical application. Methods The chondrocytes, isolated from mini swines' ears, were mixed with an injectable biocompatible matrix(Pluronic F127) to make the cell suspensions with the densities of 10,20,30,40,50,60,70×10 6/ml.The chondrocyte polymer complex was injected into the subcutaneous tissue of the swines' abdomens. Each specimen was harvested and evaluated with body mass, histological examination, and glycosaminoglycan content and type Ⅱ collagen tests after 6 weeks in vivo. Results The histological examination had showed that the neo cartilage was solid, homogenous cartilage when using 50 million chondrocytess/cc for 6 weeks. The samples with the 10 and 30 million chondrocytes/cc showed that the area of the cartilage was incomplete and separated by the remnant polymer. The mass of the samples was ranged from 30～110 mg after 6 weeks. The glycosaminoglycan content was lower from 5 8 to 9 0 percent, compared to the 9 2 percent of the normal auricular cartilage. Western Blot had presented the type Ⅱ collagen in all samples. Conclusion\ This study has demonstrated that the qutologous cartilage could be generated by using tissue engineering technique, with the histological characteristics similar to natural cartilage. Fifty million chondrocytes per cc could yield the best quality cartilage in 6 weeks.;

关 键 词：软骨细胞 软骨组织 组织工程 生物材料 软骨修复 动物实验 

分 类 号：R318.08[医药卫生—生物医学工程]