Department of Endodontics, Dental Research Center, Hamadan University of Medical Sciences, Hamadan, Iran.
Department of Endodontics, Dental School, Hamadan University of Medical Sciences, Hamadan, Iran.
Mol Biol Rep. 2023 Nov;50(11):8959-8969. doi: 10.1007/s11033-023-08747-0. Epub 2023 Sep 15.
An experimental study was conducted to examine whether melatonin influences osteogenic/odontogenic differentiation of human stem cells derived from the apical papilla (hSCAPs).
In order to isolate hSCAPs, the undeveloped root of a third molar of a human tooth was used. Melatonin was administered to the experimental groups in an osteogenic medium. No treatment was administered to the control group. The methyl thiazolyl tetrazolium (MTT) assay was performed on days 1, 2, and 3 to assess cell viability (n = 8). A determination of odontogenic/osteogenic differentiation was accomplished using alkaline phosphatase (ALP) activity alizarin red staining (ARS) (n = 6), and the expression of osteogenic genes by real-time polymerase chain reaction (RT-PCR) (n = 3) on days 1, 2, and 7. Evaluation of the data was conducted using SPSS version 18. All experiments were conducted at least three times. The Mann Whitney U test, the ANOVA analysis, Tukey's test, and t-test was implemented to analyze the data (α = 0.05).
After 24 h, 48 h, and 72 h, No significant difference was observed between the control group and the melatonin treatment group in terms of viability of hSCAPs. (from 1 up to 10 µg/ml) (P > 0.05). The assessment of ARS and ALP activity showed that melatonin treatment enhanced osteogenic differentiation of hSCAPs (P < 0.001). Melatonin treatment caused hSCAPs to show an increase of genes related to osteogenic/odontogenic differentiation. These genes included ALP, dentin sialophosphoprotein (DSPP), dentin matrix protein 1 (DMP-1), and bone sialoprotein (BSP) (P < 0.001).
Melatonin treatment enhanced osteogenic/odontogenic differentiation of hSCAPs with a dose dependent effect on cell viability.
本实验旨在研究褪黑素是否影响人根尖乳头干细胞（hSCAPs）的成骨/成牙分化。
采用未发育的第三磨牙牙根分离 hSCAPs。实验组在成骨培养基中加入褪黑素，对照组不做任何处理。在第 1、2 和 3 天用噻唑蓝（MTT）法检测细胞活力（n=8）。用碱性磷酸酶（ALP）活性茜素红染色（ARS）（n=6）和实时聚合酶链反应（RT-PCR）（n=3）检测第 1、2 和 7 天的成牙/成骨分化。采用 SPSS 18 版进行数据评估。所有实验至少进行 3 次。采用 Mann-Whitney U 检验、方差分析、Tukey 检验和 t 检验进行数据分析（α=0.05）。
在 24、48 和 72 h，hSCAPs 活力方面，对照组和褪黑素处理组之间无显著差异（1-10μg/ml）（P＞0.05）。ARS 和 ALP 活性评估表明，褪黑素处理增强了 hSCAPs 的成骨分化（P＜0.001）。褪黑素处理导致 hSCAPs 中与成骨/成牙分化相关的基因增加。这些基因包括碱性磷酸酶（ALP）、牙本质涎磷蛋白（DSPP）、牙本质基质蛋白 1（DMP-1）和骨涎蛋白（BSP）（P＜0.001）。
褪黑素处理以剂量依赖的方式增强 hSCAPs 的成骨/成牙分化，对细胞活力无影响。
