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Combined effects of simvastatin and enamel matrix derivative on odontoblastic differentiation of human dental pulp cells

机译:辛伐他汀与搪瓷基质衍生物联合作用对人牙髓细胞牙本质成牙细胞分化的影响

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Introduction: We previously reported that simvastatin and enamel matrix derivative (EMD) have a dentinogenic effect. However, there is little information about the combined effects of these 2 agents on odontoblastic differentiation. The aim of this study was to investigate the effects of combined treatment with simvastatin and EMD on odontoblastic differentiation of human dental pulp cells (hDPCs). This study further explored the role of extracellular signal-regulated kinase (ERK) as a target and mediator of the differentiation induced by simvastatin in hDPCs. Methods: The odontoblastic differentiation was analyzed by alkaline phosphatase activity, real-time polymerase chain reaction (PCR) for odontoblastic/osteoblastic markers (ie, dentin sialophosphoprotein, dentin matrix protein 1, and osteonectin), and alizarin red S staining. We also explored the role of ERK signaling as a mediator of simvastatin by Western blotting and real-time PCR. The expression of osteoblast-specific transcription factors was detected by reverse-transcription PCR. Results: The alkaline phosphatase activity and the expression of odontoblastic markers (ie, dentin sialophosphoprotein and dentin matrix protein 1) increased in simvastatin/EMD-treated cells. Mineralized nodule formation increased in EMD- and simvastatin/EMD-treated cells. Notably, the combined use of both simvastatin and EMD resulted in more potent differentiation than that observed after a single therapy. Simvastatin activated ERK phosphorylation and treatment with ERK inhibitor blocked the messenger RNA expression of odontoblastic markers. However, in simvastatin/EMD-treated cells, the expression of these genes did not decrease significantly. Compared with other groups, the EMD- and simvastatin/EMD-treated group showed a greater expression of osterix. Conclusions: Simvastatin promotes odontoblastic differentiation of hDPCs via the ERK signaling pathway. In addition, simvastatin-induced differentiation is facilitated by co-treatment with EMD. Collectively, these results suggest a new strategy to induce odontoblastic differentiation of hDPCs. ? 2013 American Association of Endodontists.
机译:简介:我们先前曾报道辛伐他汀和搪瓷基质衍生物(EMD)具有促牙本质作用。但是,关于这两种药物对牙本质母细胞分化的综合作用的信息很少。这项研究的目的是研究辛伐他汀和EMD联合治疗对人牙髓细胞(hDPCs)牙本质成骨细胞分化的影响。这项研究进一步探讨了细胞外信号调节激酶(ERK)作为辛伐他汀诱导的hDPCs分化的靶标和介体的作用。方法:通过碱性磷酸酶活性,实时聚合酶链反应(PCR)分析牙本质/牙本质标志物(即牙本质唾液磷蛋白,牙本质基质蛋白1和骨连接蛋白)和茜素红S染色,分析牙本质分化。我们还通过蛋白质印迹和实时PCR探索了ERK信号作为辛伐他汀的介质的作用。通过逆转录PCR检测成骨细胞特异性转录因子的表达。结果:在辛伐他汀/ EMD处理的细胞中,碱性磷酸酶活性和成牙母细胞标记物(即牙本质唾液磷蛋白和牙本质基质蛋白1)的表达增加。在EMD和辛伐他汀/ EMD处理的细胞中,矿化的结节形成增加。值得注意的是,辛伐他汀和EMD的联合使用比单独治疗后可产生更强的分化。辛伐他汀激活ERK磷酸化,并用ERK抑制剂处理可阻断牙本质标记的信使RNA表达。但是,在辛伐他汀/ EMD处理的细胞中,这些基因的表达没有明显降低。与其他组相比,EMD和辛伐他汀/ EMD治疗组的osterix表达更高。结论:辛伐他汀通过ERK信号通路促进hDPC的成牙本质细胞分化。此外,辛伐他汀诱导的分化可通过与EMD共同治疗而得到促进。总的来说,这些结果表明了一种诱导hDPC的齿质分化的新策略。 ? 2013美国牙医学院会员协会。

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