【摘要】 目的观察成骨生长肽(OGP)对体外人牙髓细胞增殖和矿化作用的影响。方法酶消化组织块法体外培养人牙髓细胞(hDPCs),倒置相差显微镜观察hDPCs的形态。CCK-8、碱性磷酸酶(ALP)试剂盒检测筛选最佳浓度OGP;最佳浓度组、对照组、单纯矿化液组和含最佳浓度的矿化液组做茜素红、Von Kossa染色和反转录聚合酶链反应(RT-PCR)检测。结果酶消化组织块法可以良好的体外培养hDPCs;OGP浓度在10-7～10-11mol/L均能促进hDPCs增殖和ALP活性:促增殖最佳浓度为10-10mol/L,随时间延长,与10-9mol/L浓度组间差异无统计学意义;OGP浓度为10-9mol/L时能显著增强ALP活性。茜素红和Von Kossa染色观察第14、21天,四组均有钙化结节形成,联合组结节最大,数量最多;RT-PCR结果显示第7、14、21天,联合组的Ⅰ型胶原、牙本质涎磷蛋白(DSPP)和骨钙素(OCN)的mRNA表达量均高于其他各组(P<0.01)。结论 OGP能促进体外hDPCs的增殖和分化,结合两者最优浓度为10-9mol/L;茜素红和Von Kossa染色可以观察钙化结节形成过程;OGP联合矿化诱导液可以显著提高体外hDPCs的矿化。更多还原

【Abstract】 Objective To investigate the effect of osteogenic growth peptide on proliferation and mineralization of human dental pulp cells(hDPCs) in vitro.Methods The hDPCs were cultured in vitro with tissue-explants enzymatic digestion method.The morphological characters of the cells were observed by inverted microscope.We selected the best concentration of OGP beneficial to the proliferation and ALP activity of cultured hDPCs by using CCK-8 method and alkaline phosphatase activity test;Alizarin bordeaux and Von Kossa staining methods were used to observe the minera-lization nodules.RT-PCR test was used to detect relative RNA level of Collagen Ⅰ,DSPP and OCN.Results Tissue-explants enzymatic digestion method could cultivate hDPCs well in vitro;the maximum response was obtained with 10-10 mol/L and 10-9 mol/L by using CCK-8 method.However,the optimal concentration was 10-9 mol/L by using alkaline phosphatase activity test.Alizarin bordeaux and Von Kossa staining cells cultured for 14 and 21 d,we discovered mineralization nodules in each group,but in the join group the nodules were more numerouse and larger.The results of RT-PCR test showed that mRNA level of Collagen Ⅰ,DSPP and OCN was higher in the join group.Conclusions OGP could promote the proliferation and mineralization of hDPCs in vitro,and the optimal concentration was 10-9 mol/L when considering the two aspects.Alizarin bordeaux and Von Kossa staining methods could observe the mineralization nodules dramaticlly.Mineralization induced liquid containing OGP group could markedly improve the mineralization of hDPCs in vitro.更多还原