生物蛋白胶双相接种法促进组织工程骨构建的观察  

Enhancement of tissue-engineered bone construction by bioprotein biphasic seeding technique

作　　者：刘济学 罗飞[2] 陈青 许建中[2] 

机构地区：[1]西南电子电信研究所卫生科,四川省成都市610041 [2]解放军第三军医大学西南医院骨科,重庆市400038

出　　处：《中国组织工程研究与临床康复》2009年第8期1445-1449,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家高技术研究发展计划(863)重大专项(2006AA02A122);重庆市科技攻关项目(2006AB5008)~~

摘　　要：背景:很多学者采用生物蛋白胶修饰无机支架材料改善其细胞黏附能力,并证实该方法能有效促进骨髓间充质干细胞与支架材料黏附。目的:观察采用生物蛋白胶双相接种技术促进组织工程骨体外构建的可行性。设计、时间及地点:细胞生物学水平实验,2003-01/2008-10在解放军第三军医大学组织工程研发中心实验室完成。材料:医用生物蛋白胶为广州倍特生物技术有限公司产品,同种异体冻干脱钙骨基质由解放军第三军医大学西南医院骨组织库提供。方法:以生物蛋白胶为介质,用双相接种技术将羊骨髓间充质干细胞与同种异体冻干脱钙骨基质复合,在体外构建组织工程骨,以静置接种法作为对照。主要观察指标:通过细胞计数及MTT观察不同接种密度生物蛋白胶双相接种法所构建的组织工程骨细胞数量,并通过相差显微镜和扫描电微观察细胞在支架材料中的分布情况。结果:双相接种法在不同接种密度时都可保持细胞黏附率维持在80%左右,上架细胞的数量随接种细胞密度增加成比例增加;生物蛋白胶对细胞的增殖没有明显影响,提高接种密度使细胞生长曲线提前达到平台期,各组的细胞数量峰值接近。结论:双相接种技术可显著提高种子细胞与支架材料的复合数量、复合效率并且分布均匀,是一种工程骨快速高效组织的构建方法。BACKGROUND： Bioprotein gum is frequently used to modify inorganic scaffold materials so as to improve cell adhesion ability, and it effectively promotes adhesion between bone marrow mesenchymal stem cells and scaffold materials. OBJECTIVE： To study the feasibility of bioprotein biphasic seeding technique for enhancing tissue-engineered bone construction in vitro. DESIGN, TIME AND SETTING： Cytobiological study was performed at the Laboratory of Tissue Engineering Research and Development Center, the Third Military Medical University of Chinese PLA between January 2003 and October 2008. MATERIALS： Medical bioprotein gum was provided by Beite Biotechnology Co., Ltd., Guangzhou; allogenic decalcified bone matrix was provided by bone tissue bank, Southwest Hospital of the Third Military Medical University of Chinese PLA. METHODS： By the mediation of bioprotein gum, goat bone marrow mesenchymal stem cells were compounded with allogenic decalcified bone matrix using biphasic seeding technique, Thereafter, tissue-engineered bones were constructed in vitro which were considered as the controls following rest inoculation. MAIN OUTCOME MEASURES： Numbers of tissue-engineered bones which were constructed using biphasic seeding technique were measured using cell counting and MTT method; cell distribution in scaffold materials was observed under phase contrast microscopy and scanning electron microscopy. RESULTS： With the biphasic seeding technique, the adhesive rates of different seeding densities were kept around 80%, cells upon the scaffolds increased following the increase of seeding densities. By using fibrin glue, bone marrow mesenchymal stem cells lived and proliferated well. The higher the seeding densities were, and the earlier the growth curves arrived at platform phase. All peaks were similar. CONCLUSION： Biphasic seeding technique can significantly increase the adhesive number and adhesive rate of bone marrow mesenchymal stem cells on the scaffolds, and maybe become a suitable method of constructing

关 键 词：组织工程 双相接种技术 生物蛋白胶 间充质干细胞 

分 类 号：R318[医药卫生—生物医学工程]