Demineralized Dentin Matrix Induces Odontoblastic Differentiation of Dental Pulp Stem Cells

Abstract

The aim of this study was to investigate the effect of demineralized dentin matrix (DDM) on dental pulp stem cells (DPSCs) and the potential of complexes with DPSCs and DDM for mineralized tissue formation. Stem cells derived from the dental pulp of healthy pigs aged 18 months were isolated and cultured. DPSCs were incubated with alpha-minimum essential medium treated with DDM extract at 1 mg/ml (DDM1) or 10 mg/ml (DDM10). The concentrations of 3 growth factors in DDM extract was measured by enzyme-linked immunosorbent assay. Adhesion of DPSCs on DDM and hydroxyapatite-tricalcium phosphate (HA-TCP) surfaces was observed using scanning electron microscopy. Cell proliferation was evaluated with cell counting kit-8 and migration by Transwell migration assays. Odontoblastic differentiation was assessed by alkaline phosphatase (ALP) and alizarin red staining, ALP activity and real-time polymerase chain reaction analysis of markers of ALP, runt-related transcription factor 2, type I collagen, dentin matrix acidic phosphoprotein-1, osteonectin and dentin sialophosphoprotein (DSPP). Finally, DPSCs were combined with DDM and placed subcutaneously in nude mice for 12 weeks; DPSCs combined with HA-TCP and DDM alone served as controls. DDM could promote DPSC adhesion, migration and odontoblastic differentiation. Mineralized tissue formation was observed with the DPSC and DDM combination and the DPSC and HA-TCP combination. The mineralized tissue of the DPSC + DDM combination stained positive for DSPP, similar to the dentin tissue. These results indicate that DDM induces DPSC odontoblastic differentiation, suggesting applications for dentin regeneration.

Retracted: The Role of MicroRNA‐143‐5p in the Differentiation of Dental Pulp Stem Cells into Odontoblasts by Targeting Runx2 via the OPG/RANKL Signaling Pathway

“…This finding was in concert with that of a previously conducted study, which demonstrated that a new direct pulp‐capping material containing an antibacterial resin monomer as well as Portland cement could potentially enhance the adhesion and migration of DPSCs [Yu et al, ]. A demineralized dentin matrix has displayed its ability in enhancing DPSC adhesion and migration [Liu et al, ]. SOX2 has been shown to promote the proliferation, migration and adhesion of DPSCs, evidence of this was elucidated using similar parameters and mechanisms demonstrated in our study [Liu et al, ].…”

Analysing the bioactive makeup of demineralised dentine matrix on bone marrow mesenchymal stem cells for enhanced bone repair

“…However, at low concentrations of 0.1 µg/mL, DDM presented a chemo-attractant role, inducing migration of cells through a collagen matrix, whilst 10 µg/mL inhibited cell migration. Similar actions for DDM preparations have been noted for dental pulp stem cells (Lee et al, 2015;Gonçalves et al, 2016;Liu et al, 2016) although it should be noted that for these studies the demineralised dentine extract was prepared using different protocols that varied around the theme of demineralising the mineralised tissue and thus would lead to DDM of different composition to that identified in the present study. As a direct comparison, for DDM prepared following the same protocol (Lee et al, 2015), the up-regulation of bone forming markers in dental pulp stem cells has been shown to be induced with DDM at 1 µg/ mL, with 10 µg/mL DDM significantly increasing cell proliferation, thus highlighting differential effects of DDM on cells from different tissue sources.…”

“…Placement of dentine in vivo has been reported to stimulate reparative bone formation in rabbit critical defects (Gomes et al, 2008) and in rat tooth sockets (de Oliveira et al, 2013). In vitro, DDM has been demonstrated to induce mineralisation and increased expression of bone forming markers of human DPSCs (Lee et al, 2015;Gonçalves et al, 2016;Liu et al, 2016) and murine bone marrow MSCs (BMMSCs) (Yu et al, 2014). Proteomic studies of human dentine have identified the presence of up to 289 different components, with TGF-β1 identified as a prominent growth factor (Jágr et al, 2012;Chun et al, 2011;Park et al, 2009).…”

Effect of grape seed extract (GSE) on functional activity and mineralization of OD-21 and MDPC-23 cell lines

“…ALP plays a special role in the mineralization of bone and dentin matrix, by exhibiting gradual increase at the beginning of odontoblastic differentiation. 32 In our study, GSE did not interfere in the secretion of ALP in either cell line investigated, and could be evidenced by staining in culture wells. In addition, the extract increased ALP deposition significantly in both cell lines.…”