Asgary Saeed, Nazarian Hamid, Khojasteh Arash, Shokouhinejad Noushin
Iranian Center for Endodontic Research, Research Institute of Dental Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Dental Research Center, Research Institute of Dental Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Biology and Anatomical Sciences, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
J Endod. 2014 Mar;40(3):387-92. doi: 10.1016/j.joen.2013.09.017. Epub 2013 Dec 15.
Mineral trioxide aggregate (MTA) and calcium-enriched mixture (CEM) have shown osteogenic/cementogenic/dentinogenic activities; however, their mechanism of action is not fully understood. We aimed to evaluate the effect of these biomaterials on odontogenic differentiation of human dental pulp stem cells (DPSCs).
Flow cytometry with stem cell markers for the confirmation of stemness and homogeneity was first performed. Then isolated DPSCs were seeded on prepared discs of MTA, CEM, differentiation medium (DM), and growth medium (GM) and incubated up to 14 days. Concentrations of transforming growth factor-β1, bone morphogenetic protein (BMP)2, BMP4, and fibroblast growth factor 4 were measured at each interval using an enzyme-linked immunosorbent assay reader. Gene expression of dentin sialophosphoprotein, dentin matrix protein 1, and the cytokines were evaluated by reverse-transcription polymerase chain reaction. To evaluate the cell morphology, scanning electron micrographs were taken; mineralization potential was evaluated using alizarin red S staining.
Scanning electron micrographs showed that DPSCs spread/adhered/proliferated similarly on MTA and CEM. On day 14, alizarin red S staining confirmed that mineralization occurred in all groups except GM. Expressions of dentin matrix protein 1 and dentin sialophosphoprotein genes were similar in the CEM, MTA, and DM groups; they were significantly higher compared with the GM group (P < .05). A greater amount of transforming growth factor-β1 gene was expressed in MTA compared with the other groups (P < .05). However, the expression of fibroblast growth factor 4 and BMP2 genes was significantly greater in the CEM group (P < .05). In all the tested groups, the expression of BMP4 was less than GM (P < .01); however, CEM and DM were similar but more than MTA (P < .05). Concentrations of protein product detected using an enzyme-linked immunosorbent assay reader confirmed these gene expressions.
MTA and CEM can induce osteo-/odontogenic-like phenotype differentiation of human DPSCs; however, they stimulate different gene expressions and growth factor release.
三氧化矿物凝聚体（MTA）和富钙混合物（CEM）已显示出成骨/成牙骨质/成牙本质活性；然而，它们的作用机制尚未完全明确。我们旨在评估这些生物材料对人牙髓干细胞（DPSC）牙源性分化的影响。
首先进行流式细胞术检测干细胞标志物以确认干性和同质性。然后将分离的DPSC接种到制备好的MTA、CEM、分化培养基（DM）和生长培养基（GM）圆盘上，培养14天。每隔一段时间使用酶联免疫吸附测定仪测量转化生长因子-β1、骨形态发生蛋白（BMP）2、BMP4和成纤维细胞生长因子4的浓度。通过逆转录聚合酶链反应评估牙本质涎磷蛋白、牙本质基质蛋白1和细胞因子的基因表达。为评估细胞形态，拍摄扫描电子显微镜照片；使用茜素红S染色评估矿化潜力。
扫描电子显微镜照片显示，DPSC在MTA和CEM上的铺展/黏附/增殖情况相似。在第14天，茜素红S染色证实除GM组外所有组均发生矿化。牙本质基质蛋白1和牙本质涎磷蛋白基因在CEM、MTA和DM组中的表达相似；与GM组相比显著更高（P <.05）。与其他组相比，MTA中转化生长因子-β1基因的表达量更高（P <.05）。然而，成纤维细胞生长因子4和BMP2基因在CEM组中的表达显著更高（P <.05）。在所有测试组中，BMP4的表达均低于GM组（P <.01）；然而，CEM和DM组相似但高于MTA组（P <.05）。使用酶联免疫吸附测定仪检测的蛋白质产物浓度证实了这些基因表达。
MTA和CEM可诱导人DPSC向骨/牙源性样表型分化；然而，它们刺激不同的基因表达和生长因子释放。
