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

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400 μ mol·L^-1 rhBMP-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400 μ mol·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400 μ mol·L^-1 rhBMP-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200 μ mol·L^-1 rhBMP-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。

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

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-1 respectively(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-1 was 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.