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背景:重组人骨形态发生蛋白2已被广泛应用于骨组织工程治疗骨缺损或骨不连,但是直接应用外源性骨形态发生蛋白2修复骨缺损效果不理想。目的:观察转染pIRES-骨形态发生蛋白2的大鼠骨髓间充质干细胞持续合成并分泌骨形态发生蛋白2的情况。方法:全骨髓贴壁法分离培养大鼠骨髓间充质干细胞,脂质体介导下将真核表达质粒pIRES-骨形态发生蛋白2导入大鼠骨髓间充质干细胞。结果与结论:ELISA结果显示,随着pIRES-骨形态发生蛋白2转染时间的延长,大鼠骨髓间充质干细胞分泌的骨形态发生蛋白2逐渐增多,至转染后10～12d达高峰,转染后15d,仍可见较多骨形态发生蛋白2的表达。说明转染pIRES-骨形态发生蛋白2的大鼠骨髓间充质干细胞可持续稳定分泌骨形态发生蛋白2。
BACKGROUND: Recombinant human bone morphogenetic protein 2 has been widely used in the treatment of bone defects or nonunion. However, the direct use of exogenous bone morphogenetic protein 2 in the repair of bone defects is not effective. OBJECTIVE: To observe the continuous synthesis and secretion of bone morphogenetic protein 2 by rat bone marrow mesenchymal stem cells transfected with pIRES-BMP2. METHODS: Rat bone marrow mesenchymal stem cells were isolated and cultured by whole bone marrow adherent method. The eukaryotic expression plasmid pIRES-BMP2 was transfected into rat bone marrow mesenchymal stem cells by liposome. RESULTS AND CONCLUSION: The results of ELISA showed that the bone morphogenetic protein 2 secreted by rat bone marrow mesenchymal stem cells gradually increased with the time of pIRES-BMP2 transfection, reaching the peak 10-12 days after transfection, Fifteen days after transfection, more bone morphogenetic protein 2 was still expressed. It shows that bone marrow mesenchymal stem cells transfected with pIRES-BMP2 can sustainably and stably secrete bone morphogenetic protein-2.