转化生长因子β3联合肝素在人乳牙牙髓干细胞向成牙本质样细胞分化中的作用

发布时间：2018-06-04 10:39

本文选题：人乳牙牙髓干细胞 + 转化生长因子β　；参考：《实用医学杂志》2014年12期

【摘要】：目的:探讨转化生长因子β3(TGF-β3)诱导人乳牙牙髓干细胞(SHED)向成牙本质样细胞分化的能力。方法:采用酶消化法将人乳牙牙髓分离培养,获得SHED;对体外培养的第3代SHED分别单独加入25 ng/mL的重组TGF-β3,或与肝素联合进行培养;通过Q-PCR和Western-blotting方法,分别观察SHED表达成牙本质细胞特异性标志物-牙本质涎磷蛋白(DSPP)基因及其基因表达产物-牙本质涎蛋白(DSP)的情况;通过碱性磷酸酶(AKP)试剂盒检测SHED的AKP活性的改变;细胞爬片行免疫化学染色和茜素红染色。结果:SHED在诱导体系中生长状态良好。25 ng/mL TGF-β3联合10 U/mL肝素作用组的AKP活性明显增强,与TGF-β3单独作用组、肝素单独作用组以及对照组相比差异具有统计学意义(P0.01);TGF-β3联合肝素作用组的茜素红染色呈强阳性,Q-PCR和Western-blotting结果均显示,该组的DSPP表达在mRNA水平和蛋白质水平上均明显升高。结论:在TGF-β3与肝素联合作用的诱导下,可促进SHED分化为成牙本质样细胞。
[Abstract]:Aim: to investigate the ability of transforming growth factor 尾 3 (TGF- 尾 3) to induce the differentiation of human deciduous dental pulp stem cells (SHED) into odontoblast like cells. Methods: the pulp of human deciduous teeth was isolated and cultured by enzyme digestion, and then the third generation SHED was cultured separately with 25 ng/mL recombinant TGF- 尾 3, or in combination with heparin. Q-PCR and Western-blotting methods were used. The expression of dentin sialophosphate protein (DSPP) gene and its gene expression product, dentine sialoprotein (DSPP), were observed by SHED, and the changes of AKP activity of SHED were detected by alkaline phosphatase (ALP) kit. The cells were stained with immunochemical staining and alizarin red staining. Results the AKP activity of the 10 U/mL heparin treated with 10 U/mL heparin was significantly increased in the growth state of 1: SHED. 25 ng/mL TGF- 尾 3 and TGF- 尾 3 alone. There was significant difference between heparin alone group and control group. The results of alizarin red staining and Q-PCR and Western-blotting showed that the expression of DSPP in the group treated with heparin alone was significantly higher than that in the group treated with heparin. The expression of DSPP in the group was significantly higher than that in the group treated with heparin at the level of mRNA and protein. Conclusion: TGF- 尾 3 combined with heparin can promote the differentiation of SHED into odontoid cells.
【作者单位】：广东省口腔医院南方医科大学附属口腔医院;广东省珠海市口腔医院;中山大学孙逸仙纪念医院;
【分类号】：R780.2

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