Prucksakorn Thitapat, Mutirangura Apiwat, Pavasant Prasit, Subbalekha Keskanya
Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand.
Center of Excellence in Molecular Genetics of Cancer and Human Diseases, Department of Anatomy, Faculyzety of Medicine, Chulalongkorn University, Bangkok, Thailand.
Int Dent J. 2025 Apr;75(2):1269-1276. doi: 10.1016/j.identj.2024.09.009. Epub 2024 Oct 5.
Long interspersed nuclear element-1 (LINE-1) and Alu elements are major targets of methylation, an epigenetic mechanism that is associated with several biological processes. Alterations of methylation of LINE-1 and Alu have been reported in cancers, diseases, and ageing. However, these alterations have not been studied in osteogenic differentiation of dental pulp stem cells (DPSCs), which are a promising source of tissue regeneration.
This study was performed to investigate the methylation level of LINE-1 and Alu in dental pulp stem cell-derived osteoblasts (DPSC-DOs). By using the combined bisulfite restriction analysis, the levels of total methylation and 4 patterns of methylated cytosine-phosphate-guanine (CpG) dinucleotides of LINE-1 and Alu were compared between DPSC-DOs and DPSCs.
The levels of total methylation and hypermethylated CpG dinucleotides of LINE-1 were significantly lower (P = .015 and .021, respectively), whilst levels of one pattern of partial methylated CpG dinucleotides were significantly higher in DPSC-DOs than DPSCs (P = .021). The methylation of Alu was not significantly different between DPSCs and DPSC-DOs.
Methylation alterations of LINE-1 but not Alu were found in osteogenic differentiation of DPSCs. The results of this study offer foundational insights into osteoblast differentiation from an epigenetic perspective and may contribute to advancements in bone regeneration therapy in the future.
长散在核元件1（LINE-1）和Alu元件是甲基化的主要靶点，甲基化是一种与多种生物学过程相关的表观遗传机制。LINE-1和Alu甲基化的改变已在癌症、疾病和衰老中被报道。然而，这些改变尚未在牙髓干细胞（DPSC）的成骨分化中进行研究，牙髓干细胞是组织再生的一个有前景的来源。
本研究旨在调查牙髓干细胞来源的成骨细胞（DPSC-DO）中LINE-1和Alu的甲基化水平。通过使用联合亚硫酸氢盐限制性分析，比较了DPSC-DO和DPSC中LINE-1和Alu的总甲基化水平以及4种甲基化胞嘧啶-磷酸-鸟嘌呤（CpG）二核苷酸模式。
LINE-1的总甲基化水平和高甲基化CpG二核苷酸水平在DPSC-DO中显著低于DPSC（分别为P = 0.015和0.021），而一种部分甲基化CpG二核苷酸模式在DPSC-DO中的水平显著高于DPSC（P = 0.021）。DPSC和DPSC-DO之间Alu的甲基化没有显著差异。
在DPSC的成骨分化中发现了LINE-1而非Alu的甲基化改变。本研究结果从表观遗传学角度为成骨细胞分化提供了基础见解，并可能有助于未来骨再生治疗的进展。
