摘要
目的:应用纤维蛋白胶、异种无机骨构建骨组织工程复合支架材料,立体培养兔骨髓基质细胞,探讨这种复合支架材料对种子细胞的黏附作用。方法:实验于2005-01/2006-04在吉林大学中日联谊医院卫生部创伤骨科研究室、吉林大学机械科学与工程学院工程力学系、东北师范大学遗传与细胞研究所合作完成。牛松质骨经去脂去蛋白等无机化处理后与纤维蛋白胶复合制成复合骨支架材料;将兔骨髓基质细胞作为种子细胞进行传代培养,收集后在与纤维蛋白胶、异种无机骨构成的复合支架中进行立体培养,采用相差显微镜、透射电子显微镜、苏木精-伊红染色等手段观察骨髓基质细胞在纤维蛋白胶中的生长状况。结果:①相差显微镜下观察自无机骨孔隙中见骨髓基质细胞均匀混合于纤维蛋白胶中。②细胞培养4周骨髓基质细胞形成密集立体网状。③透射电子显微镜见在复合支架中培养1个月的1个基质细胞局部有细突起,胞质内可见线粒体,核糖体,粗面内质网。结论:在纤维蛋白胶与异种无机骨构建的复合支架中骨髓基质细胞具有良好活性,可迅速扩增生长,提示这种支架材料具有适合种子细胞生长的环境。
AIM： To construct bone combined scaffolds in bone tissue engineering with fibrin glue and xenogeneic inorganic bone and perform three-dimensional culture of rabbit marrow stroma cells （MSCs） as well as probe into the adhesion effect of the scaffold on seed cells. METHODS.; The experiment was co-conducted by the Department of Orthopedics and Trauma of China-Japan Union Hospital and the Department of Mechanical Engineering, College of Mechanical Engineering of Jilin University and the Department of Genetics and Cell of Northeast Normal University between January 2005 and April 2006. The bovine cancellous bone （BCB） was dealt with H2O2, NaOH and alcohol to prepare inorganic bone. After that the BCB was combined with the fibrin glue to make combined scaffolds. Rabbit MSCs were cultured in vitro and transfer, and the MSCs were collected for three-dimension culture with fibrin glue and combined scaffold made of xenogeneic inorganic bone. The cell growth in fibrin glue was examined by HE staining, phase-contrast microscope and transmission electron microscope. RESULTS： ① Observation under the phase contrast microscope showed that the MSCs could be seen evenly mixed with the fibrin glue from inorganic bone gaps. ② At the 4= week of culture, the MSCs formed into dehse stereo-net. ③ It could be found under the transmission electron microscope that there were micro-protrusion in local stroma cells at one month after the culture, and mitochondria as well as dbosome were found in the cytoplasm with rough endoplasmic reticulum. CONCLUSION： The MSCs in scaffold combined by fibrin glue and xenogeneic inorganic bone are better in activity, and they can rapidly proliferate, which indicate that this kind of scaffold fits for the growth of seed cells.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第1期52-54,I0001,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
国家自然科学基金资助项目(30570451)~~
