应用凝胶接种技术体外分层构建工程化骨软骨复合组织的初步研究  

Preparation of laminated construction of engineered osteochondral tissue with proteic-gel technology in vitro

作　　者：段小军[1] 王富友[1] 杨柳[1] 戴刚[1] 谭洪波[1] 陈光兴[1] 郭林[1] 

机构地区：[1]第三军医大学西南医院关节外科中心,重庆400038

出　　处：《第三军医大学学报》2009年第20期1969-1972,共4页Journal of Third Military Medical University

基　　金：国家高技术研究发展计划(863计划)(2006AA02A125);国家自然科学基金(30872619);全军医学科研"十一五"计划科技攻关项目(06G079);重庆市国内科技合作计划(CSTC2007AC5069);重庆市自然科学基金(CSTC2006BB5357)~~

摘　　要：目的应用组织工程原理,探索体外构建骨软骨复合组织的关键技术,为移植修复关节骨软骨组织缺损创造条件。方法采用组织分层构建策略,用兔成骨细胞和羟基磷灰石支架材料,采用凝胶接种技术,体外构建组织工程骨;用兔软骨细胞和聚乳酸/聚磷酸钙纤维支架材料,采用凝胶接种技术,体外构建组织工程软骨;体外培养48h后,利用界面间凹凸进行压配,并结合蛋白胶粘贴形成工程化骨软骨复合组织;将构建组织移植到裸鼠皮下,以相同大小无细胞复合的支架材料为对照组,术后12周取材进行病理分析。结果采用凝胶接种技术,在体外可以初步构建组织工程骨和软骨,进而构建工程化骨软骨复合组织;工程化骨软骨复合组织移植到裸鼠皮下,术后12周实验组5例样本在成骨和成软骨区域观察到成骨和成软骨表现,但缺乏正常的钙化层界面结构,而对照组5例未观察到软骨组织形成。结论采用组织分层构建策略,采用简单的压配技术和新型凝胶接种技术可以在体外初步构建工程化骨软骨复合组织。Objective To explore the key technologies for the construction of engineered osteochondral tissue in vitro with tissue engineering principles. Methods With the aid of tissue laminated construction strategy, rabbit osteoblasts and hydroxyapatite ceramic （HAC） scaffold were used to construct tissue engineered bone in vitro with proteic-gel. Rabbit chondrocytes and PLLA/CPPf scaffold were employed to construct tissue engineered cartilage in vitro with proteic-gel. After cultured for 48 h, the engineered osteochondral tissue was prepared by press gomphosis between the 2 uneven interfaces and then pasted with albumin gel. Then the constructed was transplanted into nude mouse subcutaneously. Control group was made with the same composite scaffold but without sells. In 12 weeks after operation, the transplanted engineered osteochondral tissue was taken out for pathological analysis. Results The tissue engineered bone and cartilage were primary constructed in vitro with proteic-gel. The engineered osteochondral tissue was primary constructed. After transplantion into subcutaneous tissue for 12 weeks, all samples in the experiment group displayed bone and cartilage formation in the correspond ing area, but the calcified cartilage zone was not found on the interface of the bone and cartilage. However, the cartilage formation was not observed in the control group. Conclusion The engineered osteochondral tissue is primary constructed in vitro with the tissue laminated construction strategy and proteic-gel technology.

关 键 词：组织工程 骨软骨 成骨细胞 软骨细胞 支架材料 

分 类 号：R318.08[医药卫生—生物医学工程] R322.71[医药卫生—基础医学]