Ferutinin directs dental pulp-derived stem cells towards the osteogenic lineage by epigenetically regulating canonical Wnt signaling

Abstract

Osteoporosis is a silent systemic disease that causes bone deterioration, and affects over 10 million people in the US alone. This study was undertaken to develop a potential stem cell therapy for osteoporosis. We have isolated and expanded human dental pulp-derived stem cells (DPSCs), characterized them, and confirmed their multipotential differentiation abilities. Stem cells often remain quiescent and require activation to differentiate and function. Herein, we show that ferutinin activates DPSCs by modulating the Wnt/β-catenin signaling pathway and key osteoblastsecreted proteins osteocalcin and collagen 1A1 both mRNA and protein levels. To confirm that ferutinin modulates the Wnt pathway, we inhibited glycogen synthase kinase 3 (GSK3) and found that protein expression patterns were similar to those found in ferutinin-treated DPSCs. To evaluate the role of ferutinin in epigenetic regulation of canonical Wnt signaling, the pathway molecules Wnt3a and Dvl3 were analyzed using chromatin immunoprecipitation (ChIP)quantitative PCR approaches. We confirmed that active marks of both H3K9 acetylation and H3K4 trimethylation were significantly enhanced in the promoter sites of the WNT3A and DVL3 genes

KLF2 regulates dental pulp-derived stem cell differentiation through the induction of mitophagy and altering mitochondrial metabolism

“…Increased levels of WNT5A, CTNNB1 (β-Catenin), DVL3, and LRP6 were observed during OB differentiation (up to 7 days), confirming the involvement of the Wnt pathway. These findings support our previous data showing ferutinin directs OB differentiation of DPSCs through the Wnt signaling pathway [ 4 ], and involvement of Wnt signaling-induced Warburg effect, which is an activator of OB differentiation [ 45 ]. This data also indicates possible crosstalk among autophagy, KLF2, and Wnt signaling during OB differentiation.…”

“…The human dental pulp contains precursor cells known as dental pulp stem cells (DPSC) also exhibit self‐renewal and multilineage differentiation abilities, and shares phenotypic and morphological characteristics with MSCs. Osteogenic differentiation of DPSCs can be achieved upon stimulation with a phytoestrogen, ferutinin by modulating Wnt/β-catenin signaling as recently demonstrated [ 4 ].…”

Epigenetic Regulation of Dental Pulp Stem Cell Fate

“…The osteogenic differentiation of DPSCs is also regulated by the active mark trimethylation of lysine 4 of histone 3 (H3K4me3). Ferutinin, a daucane phytoestrogen, enhances the levels of H3K4me3 and H3K9ac on the promoters of Wnt3a and DVL3 genes in DPSCs and improves the osteogenic differentiation by activating the Wnt/ β -catenin signaling pathway [ 114 ]. These results demonstrated that histone modifications, such as H3K9me3, H3K27me3, and H3K4me3, are closely related to the differentiation process of DPSCs, especially the osteo/odontogenic differentiation.…”

Transcriptional Regulation of Osteoclastogenesis: The Emerging Role of KLF2

“…Moreover, they inhibit osteoclastic function by secreting osteoprotegerin (OPG) (3). Osteoblasts regulate osteoclasts via mechanisms distinct from the classical means of osteoclastic induction; they also exert their effects via the Wnt pathway, through Semaphorin 3A signaling, and IL-34, among others (4,5). Osteocytes are bone resident cells; they are derived from osteoblasts that have become embedded in the matrix they produced.…”