Banijamali Seyedeh Niloofar, Irani Shiva, Bakhtiar Hengameh, Askarizadeh Nahid
Department of Pediatric Dentistry, Faculty of Dentistry, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Dent Res J (Isfahan). 2022 Jan 28;19:4. doi: 10.4103/1735-3327.336689. eCollection 2022.
Stem cells from human exfoliated deciduous teeth (SHEDs) can transform into odontoblasts and . The role of 1α, 25-dihydroxyvitamin D3 (1α,25 vitD3) has been reported in the mineralization of hard tissues and teeth, as well as osteoblastic differentiation. This study aimed to assess the effect of different concentrations of 1α,25 vitD3 on odontogenic differentiation of SHEDs.
In this experimental study, second-passage SHEDs were exposed to odontogenic medium along with 0, 10, 50, 100, and 150 nmol concentrations of in 1α, 25 vitD3 to determine its optimal concentration for odontogenic differentiation. The methyl thiazolyl tetrazolium (MTT) assay was performed. Odontogenic differentiation was evaluated by QRT- polymerase chain reaction for dentin matrix protein (DMP1) and dentin sialophosphoprotein (DSPP) genes. Morphology of differentiated cells was studied by Scanning Electron Microscopy. Data were analyzed using the Kruskal-Wallis, Mann-Whitney, Friedman, and Chi-square test. < 0.05 is considered statistically significant.
MTT test result showed the two groups of odontogenic medium + 10 nm 1α,25 vitD3 and odontogenic medium + 150 nm 1α,25 vitD3 provided the most suitable conditions for cell viability at 72 h. Expression of both genes significantly increased in the presence of 1α,25 vitD3 ( < 0.001). Expression of both genes was significantly higher at 14 days compared with 7 days ( < 0.01). At both time points, expression of both genes was significantly higher in the presence of 150 nm 1α,25 vitD3 compared with 10 nm ( < 0.01). The accumulation of cells with odontoblastic morphology, cell interactions, and calcifications were evident.
1α,25 vitD3 upregulates DMP1 and DSPP and results in odontogenic differentiation of SHEDs in odontogenic medium. This upregulation increases with time and by an increase in concentration of 1α,25 vitD3.
人脱落乳牙干细胞（SHEDs）可转化为成牙本质细胞。1α,25 - 二羟基维生素D3（1α,25 vitD3）在硬组织和牙齿矿化以及成骨细胞分化中的作用已有报道。本研究旨在评估不同浓度的1α,25 vitD3对SHEDs成牙分化的影响。
在本实验研究中，将第二代SHEDs暴露于成牙培养基中，并分别添加0、10、50、100和150 nmol浓度的1α,25 vitD3，以确定其促进成牙分化的最佳浓度。进行了甲基噻唑基四氮唑（MTT）测定。通过定量逆转录聚合酶链反应检测牙本质基质蛋白（DMP1）和牙本质涎磷蛋白（DSPP）基因来评估成牙分化情况。通过扫描电子显微镜研究分化细胞的形态。使用Kruskal - Wallis、Mann - Whitney、Friedman和卡方检验分析数据。P < 0.05被认为具有统计学意义。
MTT测试结果显示，成牙培养基 + 10 nmol 1α,25 vitD3组和成牙培养基 + 150 nmol 1α,25 vitD3组在72小时时为细胞活力提供了最适宜的条件。在1α,25 vitD3存在的情况下，两个基因的表达均显著增加（P < 0.001）。与7天时相比，两个基因在14天时的表达均显著更高（P < 0.01）。在两个时间点，与10 nmol相比，150 nmol 1α,25 vitD3存在时两个基因的表达均显著更高（P < 0.01）。具有成牙本质细胞形态的细胞积累、细胞间相互作用和钙化明显可见。
1α,25 vitD3上调DMP1和DSPP，并导致SHEDs在成牙培养基中发生成牙分化。这种上调随着时间和1α,25 vitD3浓度的增加而增加。
