摘要
目的:研究转化生长因子β1(TGF-β1)在骨形态发生蛋白2(BMP-2)诱导人牙周膜干细胞(h PDLSCs)成骨分化中的作用。方法:体外分离、培养h PDLSCs,经流式细胞术鉴定后随即分为4组:空白对照组、单纯BMP-2(50 ng/m L)诱导组、BMP-2+TGF-β1(20 ng/m L)联合处理组、BMP-2+LY364947(10μmol/L)联合处理组,分别于诱导培养后3、6、9、12 d时,用实时定量PCR检测各组细胞中成骨相关基因ALP、Col-1、Runx2、OCN mRNA的表达水平;诱导培养12 d后,用Western blot法检测各细胞中上述各成骨相关基因的蛋白表达水平;诱导培养28 d后,Von Kossa染色观察各组的矿化结节形成情况。结果:单纯BMP-2诱导能显著提高h PDLSCs中ALP、Col-1、Runx2、OCN mRNA及蛋白的表达水平(P<0.05),并能显著促进其矿化结节的形成。当在BMP-2诱导液中加入TGF-β1后,可显著促进BMP-2的上述效果(P<0.05);而加入LY364947后则会显著抑制BMP-2的上述效果。结论:TGF-β1可显著促进BMP-2诱导h PDLSCs的成骨分化。
AIM： To investigate the role of transforming growth factor β1（ TGF-β1） in bone morphogenetic protein 2（ BMP-2）- induce dosteogenic differentiation of periodontal ligament stem cells（ PDLSCs）. METHODS：PDLSCs of passage 2 were treated with BMP-2（ 50 ng / m L）,BMP-2（ 50 ng / m L） ＋ TGF-β1（ 20 ng / m L）,BMP-2（ 50 ng/m L） ＋ LY364947（ 10 μmol/L） respectively and cultured for 12 d. PDLSCs without treatment were served as the controls. Real- time quantitative PCR and Western blot were performed to evaluate the expression of alkaline phosphatase（ ALP）,collagen 1（ Col- 1）,Runt- related transcription factor 2（ Runx2） and osteocalcin（ OCN）. Von Kossa staining was performed to observe the formation of mineralized nodules. RESULTS： BMP-2 promoted the expression of ALP,Col-1,Runx2 and OCN,and induced the formation of mineralized nodules. TGF-β1 remarkably increased BMP-2-induced osteogenic differentiation of PDLSCs. On the contrary,LY364947 inhibited BMP-2-induced osteogenic differentiation of PDLSCs. CONCLUSION： TGF- β1 can promote BMP-2- induced osteogenic differentiation of PDLSCs.
出处
《牙体牙髓牙周病学杂志》
CAS
2016年第6期332-337,共6页
Chinese Journal of Conservative Dentistry
关键词
人牙周膜干细胞
成骨分化
转化生长因子Β1
human periodontal ligament stem cells
osteogenic differentiation
transforming growth factor 1
