Recently, the central role of microRNAs (miRNAs) and epigenetic modifiers in biological and pathological processes, such as stem cell differentiation and inflammation, has stimulated interest. In particular, their influence in dental pulp stem cell (DPSC) differentiation has been highlighted as an exciting avenue of research in the field of Regenerative Endodontics. Although specific miRNAs have been shown to be altered in expression during dental pulp mineralization and repair processes, their interaction with epigenetic modifiers, such as histone deacetylase inhibitors (HDACi) or DNA methyltransferase inhibitors (DNMTi), has not been explored. Currently available next-generation sequencing (NGS) technologies offer the potential to explore in detail the miRNA expression profile of cells, and to investigate the effects of pharmacological inhibitors such as epigenetic modifiers on this profile. This chapter describes the experimental methods required to induce mineralization of DPCs in the presence and absence of epigenetic modifiers and analyze the resulting miRNA expression profiles using RNA sequencing (RNAseq), with a focus on bioinformatic analysis.
Keywords: DNMTi; DPSCs; Epigenetics; HDACi; Non-coding RNA; RNA sequencing; Target gene analysis; microRNA.
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.