转化生长因子β和骨形成蛋白2联合诱导小鼠成骨细胞系MC3T3-E1细胞的增殖与分化

背景:生长因子是骨组织形成、改建、修复的关键要素之一,多种生长因子联合诱导骨干细胞成骨分化相较于单一生长因子具有一定的优势,但是何种类型的联合、什么样的浓度组合才能发挥最佳的诱导作用?目前该方向研究较少.目的:探索转化生长因子β和骨形成蛋白2联合诱导对小鼠MC3T3-E1细胞增殖和分化的影响.方法:将不同质量浓度的转化...

Transforming growth factor beta combined with bone morphogenetic protein-2 induces the proliferation and differentiation of mouse MC3T3-E1 cells

BACKGROUND:Growth factors are one of the key elements for bone formation,remodeling and repair.The combination of multiple growth factors to induce osteogenic differentiation of bone stem cells has certain advantages compared with single ones.However,what combination of concentrations can play the best inducing effect has not been fully documented.OBJECTIVE:To explore the effects of transforming growth factor-βcombined with bone morphogenetic protein-2 on the proliferation and differentiation of mouse MC3T3-E1 cells.METHODS:Different concentrations of transforming growth factor-βand bone morphogenetic protein-2 were combined to cultivate MC3T3-E1 cells.The cells were then divided into six groups,including one control group and five experimental groups,and were tested at different time points.The proliferation of the cells w as determined by MTT method.Real-time quantitative RT-PCR was used to detect Runx2 and osteocalcin gene expression.Alkaline phosphatase staining was used to observe the viability and function of osteoblasts.Alizarin red staining was used to observe the amount of calcium nodules.RESULTS AND CONCLUSION:Whether transforming growth factor-βand bone morphogenetic protein-2 were used alone or in combination could significantly promote the proliferation of MC3T3-E1cells,promote the expression of Runx2 and osteocalcin,and increase the alkaline phosphatase activity and mineralization ability(P<0.05).The osteogenic effect of two growth factors in combination was positively correlated with the concentration.Even the combination at a low concentration can achieve or surpass the effect of single use.The optimal combined concentration was 1μg/L for transforming growth factor-βand 100μg/L for bone morphogenetic protein-2.Therefore,transforming growth factor-βand bone morphogenetic protein-2 used alone or in combination can significantly promote the proliferation and osteogenesis of osteoblasts,which focus on different aspects and interact with each other at different stages of proliferation and differentiation.Within the concentration range of this study,1μg/L transforming growth factor-βplus 100μg/L bone morphogenetic protein-2 show the best promotion effect.