不同因子对大鼠牙源性间充质细胞成牙本质分化的影响

发布时间：2018-08-31 20:30

【摘要】：目的:探讨b FGF、IGF1、BMP4、TGF-β1联合应用对大鼠牙源性间充质细胞(rat dental mesenchymal cells,r DMCs)成牙本质分化的影响。方法 :应用酶消化法和差速消化法分离、培养大鼠牙源性上皮细胞(r DECs)和r DMCs,经cytokeratin、vimentin免疫荧光染色鉴定其来源。应用茜素红染色、Gomori钙-钴法检测矿化液诱导下r DMCs的矿化能力。应用免疫组织化学染色、图像分析、实时荧光定量PCR和Western印迹法检测在b FGF+IGF1(组1)、TGF-β1+BMP4(组2)、b FGF+IGF1+TGF-β1+BMP4(组3)分别诱导下r DMCs中DSPP、CAP、OPN、OCN的表达差异。采用SPSS14.0软件包对数据进行统计学分析。结果:成功分离培养、鉴定r DECs和r DMCs,经矿化液诱导后的r DMCs钙结节和ALP染色阳性。组1、2中,DSPP、CAP、OPN、OCN m RNA和蛋白的表达水平均显著高于空白对照组4,差异显著(P0.01),其中组1 CAP、OCN和组2 DSPP、OPN的表达水平最高。结论:r DMCs经矿化诱导后具有成骨特性。b FGF+IGF1显著促进CAP、OCN的表达,加速r DMCs向成牙骨质细胞、成骨细胞分化与牙骨质基质、骨基质的矿化;TGF-β1+BMP4显著促进DSPP、OPN的表达,加速r DMCs向成牙本质细胞、成骨细胞分化与牙本质基质、骨基质的矿化,显示其成骨趋向。4种因子联合应用,并不具备显著的协同作用。
[Abstract]:Aim: to investigate the effect of b FGF,IGF1,BMP4,TGF- 尾 1 on odontoblast differentiation of rat odontoblast (rat dental mesenchymal cells,r DMCs. Methods: cultured rat odontogenic epithelial cells (r DECs) and r DMCs,) were isolated by enzyme digestion and differential digestion. Their origin was identified by cytokeratin,vimentin immunofluorescence staining. The mineralizing ability of r DMCs induced by mineralized solution was determined by alizarin red staining method. Immunohistochemical staining, image analysis, real-time fluorescence quantitative PCR and Western blotting were used to detect the difference of DSPP,CAP,OPN,OCN expression in r DMCs induced by b FGF IGF1 (group 1), TGF- 尾 1 BMP4 (group 2) and b FGF IGF1 TGF- 尾 1 BMP4 (group 3). The data were analyzed by SPSS14.0 software package. Results: r DECs and r DMCs, were isolated and cultured successfully. R DMCs calcium nodule and ALP staining were detected by mineralized solution. The expression levels of OCN m RNA and protein in group 1 were significantly higher than those in control group 4 (P0.01). The expression level of OCN m in group 1 and group 2 was the highest. The expression level of OCN m in group 1 and group 2 was higher than that in control group (P0.01). The expression level of OCN m in group 1 and group 2 was the highest. Conclusion the osteogenic property. BFGF IGF1 of fraction r DMCs can significantly promote the expression of CAP,OCN, accelerate the differentiation of r DMCs into cementoblasts, differentiation of osteoblasts and cemental matrix, and mineralization of bone matrix TGF- 尾 1 BMP4 can significantly promote the expression of DSPP,OPN. Accelerating the differentiation of r DMCs into odontoblast, osteoblast differentiation and mineralization of dentin matrix and bone matrix showed that there was no significant synergistic effect between r DMCs and osteoblasts.
【作者单位】：解放军济南军区第401医院口腔科;解放军总医院口腔科;
【基金】：中国博士后科学基金特别资助课题(201003774)~~
【分类号】：R781

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