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  

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

作　　者：周武 王彬娉[2] 王雅雯[2] 程亚楠[2] 黄谢山[1] Zhou Wu;Wang Binping;Wang Yawen;Cheng Yanan;Huang Xieshan(Stomatology Center,Affiliated Haikou Hospital of Xiangya Medical School,Central South University,Haikou 570208,Hainan Province,China;Department of Oral Implantology,Affiliated Haikou Hospital of Xiangya Medical School,Central South University,Haikou 570208,Hainan Province,China)

机构地区：[1]中南大学湘雅医学院附属海口医院口腔医学中心口腔颌面外科,海南省海口市570208 [2]中南大学湘雅医学院附属海口医院口腔医学中心口腔特需-种植科,海南省海口市570208

出　　处：《中国组织工程研究》2021年第23期3630-3635,共6页Chinese Journal of Tissue Engineering Research

基　　金：海南省自然科学基金(20168313),项目负责人:王彬娉。

摘　　要：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生长因子β和骨形成蛋白2与小鼠MC3T3-E1细胞混合诱导培养,共设置6个组,含1个对照组和5个实验组。在不同时间点进行检测,MTT法检测细胞的增殖情况;实时定量RT-PCR检测Runx2、骨钙素基因表达;碱性磷酸酶染色观察成骨细胞活性及功能状态;茜素红染色观察钙结节形成的数量。结果与结论:①转化生长因子β和骨形成蛋白2单独或联合使用时,都可以明显促进小鼠MC3T3-E1细胞增殖,促进Runx2、骨钙素基因的表达,增加碱性磷酸酶活性及矿化能力(P<0.05);②两者联合使用时,在实验浓度范围内促成骨效果与浓度正相关,低浓度的联合能达到或优于单独使用的效果,联合质量浓度为1μg/L转化生长因子β+100μg/L骨形成蛋白2是最优组;③提示转化生长因子β和骨形成蛋白2单独或联合诱导均能明显促进小鼠MC3T3-E1细胞增殖分化及成骨作用,二者联合诱导在增殖、分化的不同阶段既各自侧重又相互协同;在此次实验浓度范围内,1μg/L转化生长因子β+100μg/L骨形成蛋白2联合组为最优促进效果。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 differ

关 键 词：MC3T3-E1细胞 转化生长因子Β 骨形成蛋白2 成骨 细胞增殖 细胞分化 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]