bFGF基因转染的骨髓间充质干细胞在珊瑚骨表面的生长特性

目的：将转染bFGF基因的骨髓间充质干细胞（BMSCs）与珊瑚骨复合培养，观察转染bFGF基因的BMSCs在珊瑚支架材料上的生长状况。方法：穿刺抽取新西兰大白兔胫骨骨髓，采用密度梯度离心法分离BMSCs．采用贴壁筛选法对分离出的BMSCs进行纯化，利用脂质体转染bFGF—pcDNA3到BMSCs。取生长良好的转染bFGF基因的BMSCs和未转染的BMSCs，分别接种于不同珊瑚表面，利用扫描电镜观察珊瑚支架上BMSCs的生长状况：采用MTT法观察细胞一支架复合培养的BMSCs增殖情况，采用SPSS10．0软件包对数据进行t检验。结果：扫描电镜观察显示．复合培养的BMSCs贴附在珊瑚上，并在材料上完全铺展，形态多样，细胞跨越微孔表面或向孔内长人，部分区域有细胞外基质形成。MTT法检测显示，细胞-支架复合培养转染组与复合培养未转染组的BMSCs增殖状况相比有统计学差异（P〈0．05），复合培养转染组，BMSCs生长增殖强于未转染组。而复合培养的转染组与单纯培养转染组BMSCs增殖相比无统计学差异（P〉0．05）。结论：转染bFGF基因的BMSCs在珊瑚支架材料上的生长状况较未转染组好，珊瑚人工骨可以作为BMSCs支架材料，用于构建组织工程骨。

The growth characteristics of BMSCs transfected with bFGF gene after seeding on nature coral in vitro

PURPOSE： To investigate the growth characteristics of bone marrow derived mesenehymal stem cells （BMSCs） transfeeted with basic fibroblast growth factor （bFGF） gene after seeding on nature coral as its scaffolds in vitro. METHODS： The bone marrow was inspired from tibias of the rabbit. Mononuelear cells were isolated and collected from the bone marrow by gradient eentrifugation in pereoll and then cultured in vitro. BMSCs were purified by keeping the adherent cells and removing non-adherent cells repeatedly, bFGF-peDNA3 gene was transfeeted into BMSCs by using Lipofeet. BMSCs transfeeted with bFGF gene and non-transfeeted with bFGF gene were seeded into the nature coral as scaffold. The appearance, proliferativity, secretion of extraeellular matrix of BMSCs and cell affinity to the scaffold were investigated via scanning electron microscope and MTT test. SPSS 10.0 software package was used for Student＇s t test. RESULTS： Scanning electron microscope showed that BMSCs could grow well on the surface of coral and grew into pore. The appearance of BMSCs were varied and some extraeellular matrix secreted by BMSCs were seen. MTT test revealed that more proliferative activity of BMSCs transfected with bFGF-pcDNA3 than that of BMSCs non-transfected with bFGF- pcDNA3 （P〈 0.05） . But no significant difference between BMSCs transfected with bFGF-pcDNA3 were seeded into the coral and only cultured in DMEM media. CONCLUSION： More proliferative activity of BMSCs transfected with bFGF gene grow better than those of non-transfected with bFGF gene seeded into coral scaffold. Nature coral is a good compatible material for BMSCs, and can be used as scaffold to construct tissue engineering bone. Supported by Medical Research Foundation of Guangdong Province （Grant No.A2002235）