碱性成纤维生长因子和人重组骨形态发生蛋白2诱导分化兔骨髓间充质干细胞(英文)

Induced differentiation of cultured rabbit mesenchymal stem cells by basic fibroblast growth factor and recombinant human bone morphogenetic protein-2

背景:骨髓间充质干细胞依赖特定的生长和分化因子能分化为骨、软骨、脂肪等不同组织。人重组骨形态发生蛋白2有明显的骨诱导作用,能诱导未分化的骨髓间充质干细胞不可逆地形成骨与软骨,并最终导致新骨生成;碱性成纤维生长因子亦参与调控骨髓间充质干细胞的软骨与骨的分化。目的:观察碱性成纤维生长因子和人重组骨形态发生蛋白2对兔骨髓间充质干细胞增殖和分化的影响,以便寻求合适的骨形成诱导方法以代替常规的成骨培养体系。设计:随机对照实验。单位:解放军第四军医大学唐都医院骨肿瘤研究所。材料:培养的兔骨髓间充质干细胞。方法:实验于2004-06/12在第四军医大学唐都医院骨肿瘤研究所暨全军骨科中心完成。①体外培养的兔骨髓间充质干细胞分别经3种不同的生长因子(100μg/L人重组骨形态发生蛋白2,100μg/L碱性成纤维生长因子,100μg/L人重组骨形态发生蛋白2加100μg/L碱性成纤维生长因子)和成骨培养体系处理。②其增殖和分化通过四甲基偶氮唑盐比色法,碱性磷酸酶、骨钙素检测和钙结节VonKossa染色观察。主要观察指标:通过检测增殖率和碱性磷酸酶活性比较不同生长因子对兔骨髓间充质干细胞增殖和分化的影响。结果:①人重组骨形态发生蛋白2能促进兔骨髓间充质干细胞的增殖和分化,特别是其分化。②碱性成纤维生长因子能刺激兔骨髓间充质干细胞的增殖,与对照组相比,细胞增值率提高近100%;虽然对兔骨髓间充质干细胞分化无明显影响,但其能促进人重组骨形态发生蛋白2对兔骨髓间充质干细胞分化。结论:碱性成纤维生长因子和人重组骨形态发生蛋白2是兔骨髓间充质干细胞有效的分化因子,两者在体外都能促进兔骨髓间充质干细胞的增殖和分化。碱性成纤维生长因子和人重组骨形态发生蛋白2能协同作用加速骨诱导及骨形成,可以用作分化骨组织工程中的种子细胞。

BACKGROUND： Mesenehymal stem cells （MSCs） are capable of differentiating into a variety of lineages, including bone. cartilage, or fat, depending on the inducing stimuli, specific growth and differentiation factors. Recombinant human hone morphogenetic protein 2 （rhBMP-2） produced by gene engineering has an obvious osteoinductive activity and can induce undifferentiated mesenchymal cells into cartilage and bone irreversibly, resuiting in new bone formation. Basic fibroblast growth factor （bFGF） modulates chondrogenic and osteogenic differentiation of MSCs. OBJECTIVE： To investigate the effect of bFGF and rhBMP-2 on the differentiation and proliferation of cultured rabbit mesenchymal stem cell in order to find out an optimal way of osteogenesis instead of conventional osteogenic supplements （OS）. DESIGN： A randomized and controlled trial. SETTING： General Institute of Orthopaedic Ontology, Tangdu Hospital, Fourth Military Medical University of Chinese PLA. MATERIALS： The subjects were rabhit mesenchymal stem cells cultured by the author. METHODS： This experiment was conducted at the Center of Orthopaedic Surgery, General Institute of Orthopaedic Oncology, Tangdu Hospital, Fourth Military Medical University of Chinese PLA from June 2004 to De- cember 2004. ①Rabbit MSCs cultured in vitro were treated with different growth factor （100 μg/L rhBMP-2, 100 μg/L bFGF , 10 μg/L rhBMP-2 and 100 μg/L bFGF and OS; ②The proliferation and differentiation of MSCs were observed through activity of MTT, expression of alkaline phosphatase（ALP）, and yon Kossa staining. MAIN OUTCOME MEASURES： the rate of proliferation and the activity of ALP. RESULTS： ①rbBMP-2 could promote the proliferation and differentiation of MSCs, especially the cell differentiation; ②bFGF could stimulate the proliferation , the cellular proliferation rate increased 100% as compared with control group, and has no effect on differentiation of MSCs, hut it could enhance effect on the cell proliferation of rhBMP-2. CONCLUSION： bFGF1 and rhBMP-2 are effective induction faetors for MSCs. Both of them can stimulate the proliferation and differentiation of MSCs in vitro, bFGF and rhBMP-2 exerted a synergetic action in speeding up the pace of osteoinduction and osteogenesis and can be used to differ- entiate seed cells for tissue engineering bone.