Division of Restorative Dentistry & Periodontology, Dublin Dental University Hospital, Trinity College Dublin, University of Dublin, D02 F859 Dublin, Ireland.
Department of Oral Sciences, Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, Dunedin 9016, New Zealand.
Int J Mol Sci. 2023 May 11;24(10):8631. doi: 10.3390/ijms24108631.
Within regenerative endodontics, exciting opportunities exist for the development of next-generation targeted biomaterials that harness epigenetic machinery, including microRNAs (miRNAs), histone acetylation, and DNA methylation, which are used to control pulpitis and to stimulate repair. Although histone deacetylase inhibitors (HDACi) and DNA methyltransferase inhibitors (DNMTi) induce mineralisation in dental pulp cell (DPC) populations, their interaction with miRNAs during DPC mineralisation is not known. Here, small RNA sequencing and bioinformatic analysis were used to establish a miRNA expression profile for mineralising DPCs in culture. Additionally, the effects of a HDACi, suberoylanilide hydroxamic acid (SAHA), and a DNMTi, 5-aza-2'-deoxycytidine (5-AZA-CdR), on miRNA expression, as well as DPC mineralisation and proliferation, were analysed. Both inhibitors increased mineralisation. However, they reduced cell growth. Epigenetically-enhanced mineralisation was accompanied by widespread changes in miRNA expression. Bioinformatic analysis identified many differentially expressed mature miRNAs that were suggested to have roles in mineralisation and stem cell differentiation, including regulation of the Wnt and MAPK pathways. Selected candidate miRNAs were demonstrated by qRT-PCR to be differentially regulated at various time points in mineralising DPC cultures treated with SAHA or 5-AZA-CdR. These data validated the RNA sequencing analysis and highlighted an increased and dynamic interaction between miRNA and epigenetic modifiers during the DPC reparative processes.
在再生牙髓学中，为开发利用表观遗传机制（包括 microRNAs（miRNAs）、组蛋白乙酰化和 DNA 甲基化）的下一代靶向生物材料提供了令人兴奋的机会，这些机制可用于控制牙髓炎和刺激修复。尽管组蛋白去乙酰化酶抑制剂（HDACi）和 DNA 甲基转移酶抑制剂（DNMTi）可诱导牙髓细胞（DPC）群体矿化，但它们在 DPC 矿化过程中与 miRNAs 的相互作用尚不清楚。在这里，使用小 RNA 测序和生物信息学分析来建立培养物中矿化 DPC 的 miRNA 表达谱。此外，分析了 HDACi 琥珀酰亚胺基羟肟酸（SAHA）和 DNMTi 5-氮杂-2'-脱氧胞苷（5-AZA-CdR）对 miRNA 表达、DPC 矿化和增殖的影响。两种抑制剂都增加了矿化。然而，它们降低了细胞生长。表观遗传增强的矿化伴随着 miRNA 表达的广泛变化。生物信息学分析鉴定了许多差异表达的成熟 miRNA，这些 miRNA 被认为在矿化和干细胞分化中具有作用，包括对 Wnt 和 MAPK 途径的调节。通过 qRT-PCR 证明，在用 SAHA 或 5-AZA-CdR 处理的矿化 DPC 培养物中，某些候选 miRNA 在不同时间点的表达存在差异。这些数据验证了 RNA 测序分析，并强调了 miRNA 和表观遗传修饰剂在 DPC 修复过程中的相互作用增加且具有动态性。
