MTA、iRoot SP和AH Plus对人牙周膜干细胞增殖及分化的影响

Effects of MTA,iRoot SP and AH Plus on proliferation and differentiation of human periodontal ligament stem cells

目的:利用分子生物学方法检测MTA、iRoot SP和AH Plus对人牙周膜干细增殖及成骨的影响。方法:利用单克隆方法培养牙周膜干细胞,MTT检测和Annexin V-FITC/PI双染分别检测MTA、iRootSP和AH Plus对人牙周膜干细胞增殖和凋亡的影响,茜素红染色、qRT-PCR检测MTA、iRoot SP和AH Plus对牙周膜干细胞成骨能力的影响。采用SPSS 21.0软件包对数据进行统计学分析。结果:MTA在24 h和48 h呈轻度毒性,72 h无毒性;AH Plus在24 h呈轻度毒性,48 h和72 h无毒性;3 d内iRoot SP相对于MTA和AH Plus对细胞的增殖活性影响最小,几乎无毒性;MTA组和iRoot SP组矿化结节显著多于AH Plus组和对照组。2组在7、14、21 d时OC、RUNX2、COL1A、ALP基因表达均显著高于对照组,且iRoot SP组矿化基因表达最高(P<0.05)。结论:在体外,硬固后的iRoot SP对人牙周膜干细胞几乎无毒性。硬固后的iRoot SP和MTA均有一定的骨诱导能力,但在相同时间内,iRoot SP的作用更明显。

PURPOSE:To explore the effects of MTA,iRoot SP and AH Plus on periodontal ligament stem cells.METHODS:The periodontal ligament stem cells were cloned by limiting dilution culture method.The effects of MTA,iRoot SP and AH Plus on proliferation and apoptosis of periodontal ligament stem cells were detected by MTT and Annexin-V-FITC/PI double staining.Alizarin and qRT-PCR were used to evaluate the effect of MTA,iRoot SP and AH plus on osteogenesis of periodontal ligament stem cells.SPSS 21.0 software package was used for statistical analysis.RESULTS:MTA showed mild toxicity at 24 and 48 hours,AH Plus showed mild toxicity at 24 h.iRoot SP was the least(P<0.05)compared to MTA and AH Plus.The effect of three kinds of materials on apoptosis of periodontal ligament stem cells gradually decreased with the prolongation of time.Compared with the control group,the three kinds of materials were toxic at 3 d,the toxicity of MTA was the strongest and the toxicity of iRoot SP was the lowest(P<0.05).Mineralization nodules in MTA and iRoot SP group were significantly higher than those in AH Plus and control group.The expression of OC,RUNX2,COL1A and ALP gene was higher at 7,14,21 d than in the control group and the expression of iRoot SP mineralization was the greatest(P<0.05).CONCLUSIONS:The hardened iRoot SP is non-toxic to human periodontal ligament stem cells. Osteogenic ability and mineralization capacity of hardened iRoot SP on human periodontal ligamentstem cells are better than MTA.