Division of Oral Medicine and Pathology, Department of Human Biology and Pathophysiology, School of Dentistry, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-Tobetsu, Hokkaido, 061-0293, Japan.
Division of Disease Control and Molecular Epidemiology, Department of Oral Growth and Development, School of Dentistry, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-Tobetsu, Hokkaido, 061-0293, Japan.
Sci Rep. 2021 Jan 20;11(1):1852. doi: 10.1038/s41598-020-79426-4.
The DNA demethylating agent, 5-Azacytidine (5Aza), and histone deacetylase inhibitor, valproic acid (Vpa), can improve the reprogramming efficiencies of pluripotent cells. This study aimed to examine the roles of 5Aza and Vpa in the dedifferentiation of epithelial cell rests of Malassez (ERM) into stem-like cells. Additionally, the ability of stem-like cells to differentiate into mesenchymal cells was evaluated. ERM was cultured in embryonic stem cell medium (ESCM) with 1 µM of 5Aza, or 2 mM of Vpa, or a combination of 5Aza and Vpa. The cells stimulated with both 5Aza and Vpa were named as progenitor-dedifferentiated into stem-like cells (Pro-DSLCs). The Pro-DSLCs cultured in ESCM alone for another week were named as DSLCs. The stem cell markers were significantly higher in the DSLCs than the controls (no additions). The mRNA and protein levels of the endothelial, mesenchymal stem, and osteogenic cell markers were significantly higher in the Pro-DSLCs and DSLCs than the controls. The combination of a demethylating agent and a deacetylated inhibitor induced the dedifferentiation of ERM into DSLCs. The Pro-DSLCs derived from ERM can be directly reprogrammed into mesenchymal-like cells without dedifferentiation into stem-like cells. Isolated ERM treated with epigenetic agents may be used for periodontal regeneration.
DNA 去甲基化剂 5-氮杂胞苷(5Aza)和组蛋白去乙酰化酶抑制剂丙戊酸(Vpa)可提高多能细胞的重编程效率。本研究旨在探讨 5Aza 和 Vpa 在牙囊细胞间充质细胞(ERM)去分化为干细胞样细胞中的作用。此外,还评估了干细胞样细胞向间充质细胞分化的能力。将 ERM 培养在含有 1µM 5Aza 的胚胎干细胞培养基(ESCM)中,或培养在含有 2mM Vpa 的 ESCM 中,或培养在含有 5Aza 和 Vpa 的 ESCM 中。用 5Aza 和 Vpa 刺激的细胞被命名为祖细胞-去分化为干细胞样细胞(Pro-DSLCs)。在 ESCM 中单独培养一周的 Pro-DSLCs 被命名为 DSLCs。DSLCs 中的干细胞标志物明显高于对照组(无添加物)。Pro-DSLCs 和 DSLCs 中的内皮细胞、间充质干细胞和成骨细胞标志物的 mRNA 和蛋白水平明显高于对照组。去甲基化剂和去乙酰化抑制剂的组合诱导 ERM 去分化为 DSLCs。源自 ERM 的 Pro-DSLCs 可以直接重编程为间充质样细胞,而无需先去分化为干细胞样细胞。用表观遗传药物处理的分离 ERM 可能用于牙周组织再生。