Direct reprogramming of epithelial cell rests of malassez into mesenchymal-like cells by epigenetic agents

Abstract

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.

Analysis of the cells isolated from epithelial cell rests of Malassez through single-cell limiting dilution

“…PDL outgrowth explant. PDL outgrowth explant culture was performed to isolate epithelial-like cells as described previously 44,45 . Two 6-month-old porcine jaws were obtained from the livestock sales division (HOKUREN Federation of Agricultural Co-operatives; Hokkaido, Japan) and transported to our laboratory on ice.…”

Epigenetics in susceptibility, progression, and diagnosis of periodontitis

“…Then, subsequent BMP2 treatment promoted osteogenic differentiation of the de-methylated cells. Co-stimulation of epithelial cell rests of Malassez with 5-aza-2`-deoxycytidine and valproic, a histone deacetylase inhibitor, was sufficient to induce the trans-differentiation into osteogenic mesenchymal-like cells [113] . These results showed that enforced epigenetic alteration is an attractive strategy to efficiently generate osteogenic cells ex vivo for regenerative medicine.…”

Application and prospects of somatic cell reprogramming technology for spinal cord injury treatment

“…Histone deacetylases play an important role in chromatin remodeling and epigenetic regulation, and their inhibitors can access chromatin to promote the transcription of silenced genes ( Zhao et al, 2020 ). VPA, a histone deacetylase inhibitor, can increase cell reprogramming efficiency and promote normal neurogenesis and the maturation of neuroblasts ( Yoshida et al, 2021 ). Studies have shown that applying the transcription factor Sox2 combined with VPA can further convert neurons into GABAergic-like neurons in vivo ( Su et al, 2014 ).…”