Abstract
To understand the functions of secretory proteins in odontogenesis and to further the understanding of the different molecular events during odontogenesis and osteogenesis, we induced the odonto/osteogenic differentiation of stem cells from dental apical papilla (SCAPs) and bone marrow–derived stem cells (BMSCs) in vitro and compared the expression of secretory proteins during early odonto/osteogenic differentiation using high-performance liquid chromatography with tandem mass spectrometry. The results revealed significant changes by at least 50% in 139 SCAP proteins and 203 BMSC proteins during differentiation. Of these, 92 were significantly upregulated and 47 were significantly downregulated during the differentiation of SCAPs. Most of these proteins showed the same trend during the differentiation of BMSCs. Among the proteins that showed significantly changes during the differentiation of SCAPs and BMSCs, we found that transforming growth factor-β2 (TGFβ2) is a key protein in the network with powerful mediation ability. TGFβ2 was secreted more by SCAPs than BMSCs, was significantly upregulated during the differentiation of SCAPs and was significantly downregulated during the differentiation of BMSCs. Furthermore, the effects of recombinant human TGFβ2 and TGFβ1 on the odonto/osteogenic differentiation of SCAPs and BMSCs were investigated. Real-time reverse transcription polymerase chain reaction (RT-PCR) and western blotting data revealed that TGFβ2 enhanced the odontogenic-related markers [dentin sialophosphoprotein (DSPP), dentin matrix protein 1 (DMP1)] and inhibited the osteogenic-related marker bone sialoprotein (BSP) in SCAPs, whereas TGFβ1 enhanced the BSP expression and inhibited the DSPP and DMP1 expression at early odonto/osteogenic differentiation of SCAPs. However, in BMSCs, TGFβ2 enhanced the expression of alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2), DSPP, and DMP1, whereas TGFβ1 enhanced the expression of ALP and RUNX2, with no significant intergroup difference of DSPP at the early odonto/osteogenic differentiation of BMSCs. TGFβ2 is a potentially important molecule with a distinct function in the regulation of odontogenesis and osteogenesis.
Highlights
The treatment of immature permanent teeth with necrotic pulp is a challenging task for dental professionals
To explore the secretory proteins during their early odonto/osteogenic differentiation, the conditioned media from SCAP, bone marrow–derived stem cells (BMSCs), SCAP-inducing, and BMSC-inducing groups were analyzed by HPLC-MS/MS (Figure 2A)
TGFβ2 and tumor necrosis factor receptor superfamily member 1A were significantly upregulated during the differentiation of SCAPs and significantly downregulated during the differentiation of BMSCs
Summary
The treatment of immature permanent teeth with necrotic pulp is a challenging task for dental professionals. During revascularization/regeneration procedures, the evoked-bleeding step leads to a substantial influx of stem cells from dental apical papilla (SCAPs) into the root canal, which contributes to root development and apical closure (Cao et al, 2015). Several histological findings reported that most tissues formed in the canal of the revitalized immature permanent tooth lack organized pulp–dentin complex and consist of ectopic bone, fibrous tissues, and cementum (Martin et al, 2012; Saoud et al, 2015; Peng et al, 2017). It is presumed that stem cells from alveolar bone would migrate into the root canal and initiate ectopic calcification deposition (Cao et al, 2015; Saoud et al, 2015). It is necessary to find some important factors to regulate the differentiation of stem cells into odontoblasts and osteoblasts
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