Abstract
BackgroundExtracellular matrix secretion and odontoblastic differentiation in human dental pulp stem cells (hDPSCs) are the cellular bases for reparative dentinogenesis. Osteomodulin (OMD) is a member of the small leucine-rich proteoglycan family distributed in the extracellular matrix but little is known about its role in osteo/odontogenic differentiation. The objective of this study was to investigate the role of OMD during osteo/odontoblastic differentiation of hDPSCs.MethodshDPSCs were selected using immune-magnetic beads and their capability of multi-differentiation was identified. OMD knockdown was achieved using short hairpin RNA (shRNA) lentivirus and was confirmed by western blot. Gene expression was measured by real-time qPCR and osteo/odontoblastic differentiation of hDPSCs was determined by alizarin red S staining.ResultsCompared with uninduced cells, the transcription of OMD was up-regulated by 35-fold at the late stage of osteo/odontogenic differentiation. shRNA-mediated gene silencing of OMD decreased the expression of odontoblastic genes, such as alkaline phosphatase (ALP), dentin matrix acidic phosphoprotein 1 (DMP1) and dentin sialophosphoprotein (DSPP). Besides, knockdown of OMD attenuated the mineralized nodules formation induced by osteo/odontogenic medium.ConclusionsThese results implied that OMD may play a pivotal role in modulating the osteo/odontoblastic differentiation of hDPSCs.
Highlights
Extracellular matrix secretion and odontoblastic differentiation in human dental pulp stem cells are the cellular bases for reparative dentinogenesis
Flow cytometric analysis was one of the methods based on cell surface molecules. human dental pulp stem cells (hDPSCs) showed the characteristic pattern of mesenchymal stem cells (MSCs)-associated surface markers, including CD73, CD90, CD105 and CD166 and were negative for hematopoietic stem cell surface markers CD34 and CD45
Calcium deposition was confirmed by alizarin red S staining (Fig. 1b) and lipid formation was revealed by oil red O staining (Fig. 1c), which indicated hDPSCs’ differentiation into cells like osteoblasts and adipocytes
Summary
Extracellular matrix secretion and odontoblastic differentiation in human dental pulp stem cells (hDPSCs) are the cellular bases for reparative dentinogenesis. Osteomodulin (OMD) is a member of the small leucine-rich proteoglycan family distributed in the extracellular matrix but little is known about its role in osteo/ odontogenic differentiation. The objective of this study was to investigate the role of OMD during osteo/ odontoblastic differentiation of hDPSCs. The dental pulp contains a unique precursor population of mesenchymal stem cells (MSCs) [1]. Cultured dental pulp stem cells can differentiate into odontoblast-like cells and form calcium. Dentinogenesis is highly regulated by the expression of the extracellular matrix (ECM) proteins. An important family of molecules with regulatory functions is the small leucine-rich proteoglycans (SLRPs), which are extensively involved in the dentinal biomineralization [8]. It has been confirmed that biglycan and decorin are identified in the matrices of dentin and implicated in dentinogenesis [9]
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