Abstract:
The utilization of canine mesenchymal stem cells (cMSCs) with multipotent capabilities has been regarded for possible therapy of incorrigible bone disease. Although various sources of cMSCs show similar characteristics, they are different in osteogenic potential due to their original cellular sources. This study was designed to globally explore and analyze the in vitro differentiation potential and behavior of canine bone-marrow derived mesenchymal stem cells (cBM-MSCs) and canine dental pulp stem cells (cDPSCs) toward osteogenic lineage. An in vitro osteogenic differentiation potential of the cells was preliminarily compared in terms of alkaline phosphatase activity assay and Von Kossa staining. Global study of an in vitro osteogenic differentiation potential of the isolated cells was performed using proteomic-based analysis through mass spectrometry with dimethyl labelling method at day 7 and 14 post-induction, comparing with undifferentiated cells. The result presented that cBMSCs and cDPSCs contained osteogenic differentiation potential but had differences in their alkaline phosphatase activity level and mineralization. Proteomics profiling revealed that cBM-MSCs and cDPSCs showed the differences in their protein expression of signaling pathways, extracellular matrix organization, cell cycle, metabolism, transport of small molecules, and vesicle-mediated transport which have been shown to involve in bone regeneration mechanisms. Basing on database analysis and functional assay confirmation, there were four potential osteogenic-regulating pathways; Wnt signaling, Notch signaling, bone-morphogenetic protein (BM-related signaling and transforming growth factor (TGF-related signaling, which played the crucial regulating of cBM-MSCs and cDPSCs toward osteogenic lineage. The obtained results could be used as a comprehensive data and principal knowledge of the osteogenic differentiation potential of cBM-MSCs and cDPSCs in vitro and the trend of MSC-based tissue engineering for osteogenic regenerative therapy, concentrating on cMSCs application.
