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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: Bone marrow mesenchymal stem cells can differentiate into different tissues such as bone, cartilage and fat depending on the specific growth and differentiation factors. Recombinant human bone morphogenetic protein 2 has obvious osteoinductive effect and can induce undifferentiated bone marrow mesenchymal stem cells to irreversibly form bone and cartilage and ultimately lead to new bone formation. Basic fibroblast growth factor is also involved in the regulation of bone marrow filling Differentiation of cartilage and bone in mesenchymal stem cells. OBJECTIVE: To observe the effects of basic fibroblast growth factor (rhFGF) and human recombinant bone morphogenetic protein-2 on the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells in order to find a suitable induction method for bone formation instead of the conventional osteogenic culture system. Design: Randomized controlled experiment. Unit: Fourth Military Medical University Tangdu Hospital Cancer Institute. Materials: Cultured rabbit bone marrow mesenchymal stem cells. METHODS: The experiment was performed at the Institute of Bone Tumor and Military Orthopedic Center, Tangdu Hospital, Fourth Military Medical University from June to December 2004. ① Rabbit bone marrow mesenchymal stem cells cultured in vitro were cultured with three different growth factors (100μg / L recombinant human bone morphogenetic protein 2, 100μg / L basic fibroblast growth factor, 100μg / L recombinant human bone morphogenetic protein 2 100μg / L basic fibroblast growth factor) and osteogenic culture system. ② The proliferation and differentiation were observed by MTT assay, alkaline phosphatase, osteocalcin test and Von Kossa staining of calcium nodules. MAIN OUTCOME MEASURES: The effects of different growth factors on the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells were compared by detecting the proliferation rate and alkaline phosphatase activity. Results: ① Recombinant human bone morphogenetic protein-2 can promote the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells, especially its differentiation. ② Basic fibroblast growth factor can stimulate the proliferation of rabbit bone marrow mesenchymal stem cells, compared with the control group, the cell proliferation rate increased by nearly 100%; Although the rabbit bone marrow mesenchymal stem cells did not significantly affect the differentiation, but it can promote human Recombinant bone morphogenetic protein 2 differentiates rabbit bone marrow mesenchymal stem cells. CONCLUSION: Basic fibroblast growth factor and human recombinant bone morphogenetic protein-2 are effective differentiation factors of rabbit bone marrow mesenchymal stem cells, both of which can promote the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells in vitro. Basic fibroblast growth factor and human recombinant bone morphogenetic protein 2 can accelerate bone induction and bone formation in synergy and can be used as seed cells in differentiated bone tissue engineering.