Abstract
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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Farges J-C, Alliot-Licht B, Renard E et al (2015) Dental pulp defence and repair mechanisms in dental caries. Mediat Inflamm 2015:1–16. https://doi.org/10.1155/2015/230251
Yang J, Yuan G, Chen Z (2016) Pulp regeneration: current approaches and future challenges. Front Physiol 7:58. https://doi.org/10.3389/fphys.2016.00058
Smith AJ, Duncan HF, Diogenes A et al (2016) Exploiting the bioactive properties of the dentin-pulp complex in regenerative endodontics. J Endod 42:47–56
Murray PE, Garcia-Godoy F, Hargreaves KM (2007) Regenerative endodontics: a review of current status and a call for action. J Endod 33:377–390. https://doi.org/10.1016/j.joen.2006.09.013
Duncan HF, Smith AJ, Fleming GJP, Cooper PR (2016) Epigenetic modulation of dental pulp stem cells: implications for regenerative endodontics. Int Endod J 49:431–446. https://doi.org/10.1111/iej.12475
Duncan HF, Smith AJ, Fleming GJP, Cooper PR (2012) Histone deacetylase inhibitors induced differentiation and accelerated mineralization of pulp-derived cells. J Endod 38:339–345. https://doi.org/10.1016/j.joen.2011.12.014
Zhang D, Li Q, Rao L et al (2015) Effect of 5-Aza-2′-deoxycytidine on odontogenic differentiation of human dental pulp cells. J Endod 41:640–645. https://doi.org/10.1016/j.joen.2014.12.006
Chuang D-M, Leng Y, Marinova Z et al (2009) Multiple roles of HDAC inhibition in neurodegenerative conditions. Trends Neurosci 32:591–601. https://doi.org/10.1016/j.tins.2009.06.002
Kearney M, Cooper PR, Smith AJ, Duncan HF (2018) Epigenetic approaches to the treatment of dental pulp inflammation and repair: opportunities and obstacles. Front Genet. https://doi.org/10.3389/fgene.2018.00311
Yang W, Harris MA, Cui Y et al (2012) Bmp2 is required for odontoblast differentiation and pulp vasculogenesis. J Dent Res 91:58. https://doi.org/10.1177/0022034511424409
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Kearney, M., Duncan, H.F. (2023). Micro-RNA Profiling in Dental Pulp Cell Cultures. In: Seymour, G.J., Cullinan, M.P., Heng, N.C., Cooper, P.R. (eds) Oral Biology. Methods in Molecular Biology, vol 2588. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2780-8_20
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2780-8_20
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2779-2
Online ISBN: 978-1-0716-2780-8
eBook Packages: Springer Protocols