Genome-wide DNA (hydroxy) methylation reveals the individual epigenetic landscape importance on osteogenic phenotype acquisition in periodontal ligament cells

Author(s)

Ferreira, Rogerio S. ^[1] ; Assis, Rahyza I. F. ^[2] ; Feltran, Georgia da S. ^[3] ; do Rosario Palma, Iasmin Caroline ^[4] ; Francoso, Beatriz G. ^[1] ; Zambuzzi, Willian F. ^[3] ; Andia, Denise C. ^[1] ; da Silva, Rodrigo A. ^{[4, 5]}

Affiliation

^[1] Univ Paulista, Hlth Sci Inst, Sch Dent, Dr Bacelar St 1212, BR-04026002 Sao Paulo - Brazil

^[2] Univ Estadual Campinas, Piracicaba Dent Sch, Dept Prosthodont & Periodont, Piracicaba - Brazil

^[3] Sao Paulo State Univ, Inst Biosci, Dept Chem & Biol Sci, Botucatu, SP - Brazil

^[4] Univ Taubate, Dept Dent, Taubate, SP - Brazil

^[5] Univ Paulista, Program Environm & Expt Pathol, Sao Paulo - Brazil

• Document type: Journal article
• Source: Journal of Periodontology; AUG 2021.
• Web of Science Citations: 0

Abstract

Background Mesenchymal cells' biology has been an important investigative tool to maximize bone regeneration through tissue engineering. Here we used mesenchymal cells from periodontal ligament (PDLCs) with high (h-) and low (l-) osteogenic potential, isolated from different donors, to investigate the impact of the individual epigenetic and transcriptional profiles on the osteogenic potential. Methods Genome-wide and gene-specific DNA (hydroxy) methylation, mRNA expression and immunofluorescence analysis were carried out in h- and l-PDLCs at DMEM (non-induced to osteogenesis) and OM (induced-3rd and 10th days of osteogenic differentiation) groups in vitro. Results Genome-wide results showed distinct epigenetic profile among PDLCs with most of the differences on 10th day of OM; DMEMs showed higher concentrations (xOM) of differentially methylated probes in gene body, intronic and open sea (3rd day), increasing this concentration in TSS200 and island regions, at 10 days. At basal levels, h- and l-PDLCs showed different transcriptional profiles; l-PDLCs demonstrated higher levels of NANOG/OCT4/SOX2, BAPX1, DNMT3A, TET1/3, and lower levels of RUNX2 transcripts, confirmed by NANOG/OCT4 and RUNX2 immunofluorescence. After osteogenic induction, the distinct transcriptional profile of multipotentiality genes was maintained among PDLCs. In l-PDLCs, the anti-correlation between DNA methylation and gene expression in RUNX2 and NANOG indicates methylation could play a role in modulating both transcripts. Conclusions Epigenetic and transcriptional distinct profiles detected at basal levels among PDLCs were maintained after osteogenic induction. We cannot discard the existence of a complex that represses osteogenesis, suggesting the individual donors' characteristics have significant impact on the osteogenic phenotype acquisition. (AU)

• FAPESP's process: 13/09650-8 - Epigenetic regulation in human mesenchymal stem cells
• Grantee: Denise Carleto Andia
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 17/12158-9 - Epigenetic regulation of osteogenic potential in mesenchymal stem cells derived from periodontal ligament
• Grantee: Rahyza Inacio Freire de Assis
• Support Opportunities: Scholarships in Brazil - Doctorate