牙本质非胶原蛋白对人根尖牙乳头干细胞增殖及分化能力的影响

摘要

AIM:To investigate the effects of dentin non-collagenous proteins (DNCPs)on the proliferation and differentiation of human stem cells from the apical papilla (hSCAPs)in vitro.METHODS:hSCAPs were treated with 10 μg/mL of DNCPs,and the proliferation of the hSCAPs was detected by MTT assay.To investigate the differen-tiation of hSCAPs,alkaline phosphotase (ALP)activity was determined by enzyme kinetics methods,the mRNA ex-pression of ALP,OCN and COL I was detected by real-time PCR,and the mineralization was detected by Alizarin red staining assay.RESULTS:The proliferation activity was not statistically different differect between the DNCPs-trea-ted,and the control cells(P >0.05).After 7 and 14 days culture in DNCPs containing medium,the mRNA expres-sions of ALP,OCN and COL I in hSCAPs were significantly upregulated(P <0.05).ALP activity of dNCPs-treated cells was significantly promoted(P <0.05).After 3 weeks,the calcified nodules in DNCPs-treated cells were bigger and more that those in the controls(P <0.05).CONCLUSION:DNCPs can promote the mineralization ability of hSCAPs,while have no significant effect on the proliferation ability.%目的：探讨牙本质非胶原蛋白（DNCPs）对人根尖牙乳头干细胞（hSCAPs）增殖及分化能力的影响。方法：hSCAPs 经 DNCPs 诱导后，用 MTT 法检测细胞增殖能力；碱性磷酸酶（ALP）试剂盒测定 ALP；RT－PCR检测 ALP、骨钙素（OCN）及 I 型胶原（Col I）的 mRNA 表达变化；茜素红矿化结节染色及定量检测DNCPs 对 hSCAPs 分化能力的影响。结果：DNCPs 诱导1、3、5、7 d 时，hSCAPs 的增殖活性与对照组相比无显著差异（P ＞0．05）；诱导7、14 d 时，hSCAPs 的 ALP、Col I 和 OCN mRNA 表达水平均显著上调（P ＜0．05）；诱导3、5、7 d 时，hSCAPs 的 ALP 活性明显上调（P ＜0．05）；诱导3周后，茜素红染色显示 DNCPs 诱导组的钙化结节形成能力明显高于对照组（P ＜0．05）。结论：DNCPs 可明显促进 hSCAPs 矿化能力，对其增殖活性的影响则不明显。