转化生长因子β3促进兔牙髓干细胞成骨向分化的体外实验研究

Transforming growth factor-β3 induces osteogenic differentiation of rabbit dental pulp stem cells in vitro

目的:探讨转化生长因子β3(TGF-β3)体外诱导兔牙髓干细胞(rDPSCs)骨向分化的效果及其作用机制。方法:酶解组织块法分离的rDPSCs分为实验组(rDPSCs+80 ng/ml TGF-β3),对照组(rDPSCs+矿化液)和空白组(rDPSCs),免疫细胞化学染色法行COL-I、OCN, Runx-2染色;RT-qPCR检测各组ALP、COL-I、OCN、Runx-2表达水平;茜素红(ARS)染色观察矿化结节。结果:免疫细胞化学染色结果示实验组成骨蛋白表达明显强于其余2 组;RT-qPCR结果显示实验组中成骨基因的表达均高于其余2 组(P<0.05);实验组矿化结节形成数高于其余2组(P<0.05)。结论:TGF-β3能促进rDPSCs骨向分化。

Objective: To investigate the effects and the mechanism of transforming growth factor-β3(TGF-β3)on the osteogenic differentiation of rabbit dental pulp stem cells(rDPSCs). Methods: rDPSCs of the third passage were divided into the experimental group(rDPSCs+80 ng/ml TGF-β3),control group(rDPSCs+osteogenic medium)and blank control group(conventional culture medium). Immunohistochemical staining(IHC) was used to observe the protein expression of COL-I,OCN and Runx-2 of the cells;RT-qPCR was carried out to measure the mRNA expression of ALP,COL-I,Runx-2 and OCN, and Alizarin Red S(ARS) staining was carried out to testify mineralized matrix productivity. Results: IHC showed that the protein expression of COL-I,Runx-2 and OCN mRNA of the experimental group was stronger than that of the other groups. RT-qPCR results revealed that the ALP,COL-I,Runx-2 and OCN mRNA expression in the experimental group was higher than that of the other groups(P<0.05). The average number of mineralized nodules in the experimental group was more than that of the other groups(P<0.05). Conclusion: TGF-β3 can promote the osteogenic differentiation of rDPSCs in vitro.