摘要
目的探讨重组人胰岛素样生长因子-I(rhIGF-I)和重组人骨形成蛋白-2(rhBMP-2)单独及联合应用对MC3T3-E1和NIH3T3细胞增殖和分化的影响。方法单独或联合应用不同浓度rhIGF-I和rhBMP-2作用于MC3T3-E1和NIH3T3细胞后,采用四甲基偶氮唑蓝(MTT)检测细胞增殖情况,碱性磷酸酶(ALP)试剂盒检测细胞分化情况,放射免疫法检测细胞分泌的骨钙素(OC)水平。结果1~75ng/mlrhIGF-I与10~100ng/mlrhBMP-2作用于MC3T3-E1和NIH3T3细胞后,均能显示明显的促细胞增殖(P<0.01)及使ALP活性增高(P<0.05)的作用。各浓度rhIGF-I或rhBMP-2对MC3T3-E1及NIH3T3细胞作用8、24和48h的MTT检测结果相似。rhIGF-I和rhBMP-2联合作用后,对两种细胞的促增殖、增强ALP活性的作用均较各自单独作用更为明显(P<0.05)。单独或联合应用不同浓度rhIGF-I和rhBMP-2对细胞分泌的OC水平均无显著影响(P>0.05)。结论rhIGF-I和rhBMP-2具有明显的促细胞增殖及早期分化的协同作用,但对成骨末期细胞分化影响不大,其活性主要与使用剂量有关。
Objective To demonstrate the effects of recombinant human insulin-like growth factor I (rhIGF-I) or recombinant human bone morphogenetic protein-2 (daBMP-2) and combined application of the two cytokines on proliferation and differentiation of MC 3T3-E1 cells and NIH 3T3 cells. Methods Mouse osteobalast-like cell MC3T3-E1 and mouse fibroblast cell N1H3T3 were treated with different dosages of rhIGF-I or rhBMP-2 and rhIGF-I plus rhBMP-2. Proliferations of the two ceils were measured by methylthiazol tetrazolium(MTT) method. Differentiations of the two cells were examined by using alkaline phosphatase( ALP) measurement kit. Radioimmunoassay was applied to detect levelsofosteocalcin(OC) secreted by the two cells. Results MC3T3-E1 cells and NIH 3T3 cells treated with 1- 75 ng/ml rhIGF-I and10- 100 ng/ml rhBMP-2 showed obvious effects of promoting proliferation ( P 〈 0.01 ) and increasing activities of cellular ALP ( P 〈 0.05). The results of MTT examinations were similar by using the same different concentrations of rhIGF-I and rhBMP-2 treated for 8, 24 and 48 h, respectively. RhIGF-I plus rhBMP-2 showed stronger effects on promoting proliferation and increasing cellular ALP activities for both the two cells than using any one of the two cytokines ( P 〈 0.05). However, no significant changes in levels of OC secreted by the two cells was found when using different dosages of rhIGF-I or rhBMP-2 and rhIGF-I plus rhBMP-2( P 〉 0,05), Conclusion rhIGF-I and rhBMP-2 had synergistieal effects on promoting cell proliferation and early cell differentiation depending on the used dosages, but they had no significant effects on promoting advanced cell differentiation.
出处
《口腔医学》
CAS
2006年第3期193-196,共4页
Stomatology