人重组骨形成蛋白-7对人牙周膜成纤维细胞增殖及分化能力的影响

Effect of recombinant human bone morphogenetic protein-7 on proliferation and differentiation of human periodontal ligament cells

目的:研究人重组骨形成蛋白-7(rhBMP-7)对体外培养的人牙周膜成纤维细胞(HPDLCs)增殖、分化能力的影响,为进一步探讨其促进牙周组织再生奠定细胞学基础。方法:选取因阻生或正畸需要拔除的牙齿,无菌条件下刮取根中1/3的牙周膜组织,采用组织块法培养牙周膜成纤维细胞,经免疫细胞化学鉴定后接种培养板,加入含浓度为50、100、200及400μmol.L-1rhBMP-7的培养液。分别于培养后1、3及5 d,采用MTT法测定HPDLCs的增殖能力,采用ELISA检测HPDLCs培养液中ALP的活性。结果:原代牙周膜细胞生长缓慢,加入rhBMP-7后细胞生长旺盛,细胞形态为纺锤形或成纤维样,胞浆透明,生长状态良好。随着时间的延长,rhBMP-7在100、200及400μmol.L-1各浓度时,HPDLCs细胞的增殖率呈逐渐增加的趋势(P<0.01),但各浓度组间比较差异无显著性(P>0.05);与对照组比较,100、200及400μmol.L-1rhBMP-7各浓度组HPDLCs细胞裂解液和培养液中的ALP活性均增高(P<0.05或P<0.01),其中200μmol.L-1rhBMP-7组ALP活性最高。结论:BMP-7可能通过促进HPDLCs细胞的增殖和向骨细胞分化功能而参与牙周组织改建。

Objective To study the effect of recombinant human bone morphogenetic protein-7（rhBMP-7） on proliferation and differentiation of human periodontal ligament cells（HPDLCs） cultivated in vitro.Methods The HPDLCs were tested Vimentin positive and Ck（pan） negative.Different dosages（50,100,200 and 400 μmol·L^-1 of rhBMP-7 were added in the cultured HPDLCs.1,3,5 d after administration of rhBMP-7,the proliferation and osteogenic differentiation of HPDLCs were examined by MTT assay and the activity of alkaline phosphatase（ALP） was detected with ELISA.Results The administration of rhBMP-7 significantly elevated the proliferation rate and differentiation of HPDLCs with the dosage of l00,200 and 400 μmol·L^-1respectively（P〈0.01）,no difference was found between different dosages groups（P〉0.05）.Moreover,rhBMP-7 significantly induced the osteogenic differentiation ability with the increased expression level of ALP and 200 μmol·L^-1was the most effective dosage in the differentiation ability.Conclusion BMP-7 may participate in the remodeling of periodontal tissue by regulating the proliferation and osteogenic differentiation of HPDLCs.