A Comparative In Vitro Analysis of the Osteogenic Potential of Human Dental Pulp Stem Cells Using Various Differentiation Conditions

Abstract

Dental pulp stem cells (DPSCs) have excellent proliferative properties, mineralization potential and can be easily obtained from third molar teeth. Recently, many studies have focused on isolation and differentiation of DPSCs. In our study, we focused on biological properties of non-differentiated DPSCs in comparison with osteogenic differentiated cells from DPSCs. We analyzed morphology as well as mineralization potential using three varied osteogenic differentiation media. After fifteen days of differentiation, calcium deposit production was observed in all three osteogenic differentiation media. However, only one osteogenic medium, without animal serum supplement, showed rapid and strong calcification—OsteoMAX-XF™ Differentiation Medium. Therefore, we examined specific surface markers, and gene and protein expression of cells differentiated in this osteogenic medium, and compared them to non-differentiated DPSCs. We proved a decrease in expression of CD9 and CD90 mesenchymal stem cell surface markers, as well as downregulation in the expression of pluripotency genes (NANOG and OCT-4) and increased levels of expression in osteogenic genes (ALP, BSP, OCN and RUNX2). Moreover, osteogenic proteins, such as BSP and OCN, were only produced in differentiated cells. Our findings confirm that carefully selected differentiation conditions for stem cells are essential for their translation into future clinical applications.

Biological Characteristics and Osteogenic Differentiation of Ovine Bone Marrow Derived Mesenchymal Stem Cells Stimulated with FGF-2 and BMP-2

“…This observation suggests that the decrease of CD90 gene expression is a result of FGF-2 and BMP-2 activity. This phenomenon may be the effect of an ongoing osteogenic differentiation of the examined ovine BM-MSCs, as demonstrated in studies on the osteogenic differentiation of human dental pulp stem cells and a decreased expression of the MSC surface marker CD90 [ 40 ]. The data showed little variation in the expression level of the negative MSC surface markers, especially HLA DR.…”

Cannabidiol and Vitamin D3 Impact on Osteogenic Differentiation of Human Dental Mesenchymal Stem Cells

“…There are successfully used several mechanisms of inducing osteogenic differentiation of DPSCs, like gene expression inhibition [ 45 ], identification of circular RNA mechanisms, which are recognized to have a crucial role in bone regeneration correlated with stem cells [ 46 ]. Okajacekova et al [ 47 ] tested the osteoblast differentiation of DPSCs by keeping them under different conditions and found that only one osteogenic medium xeno-free without animal serum supplement (OsteoMAX-XF™ Differentiation Medium) showed a rapid and strong calcification [ 47 ]. DFSCs and APSCs have also proven their osteoblast differentiation potential in different conditions, as bone regeneration on titanium implants [ 48 ] or cranial defects [ 49 ], for DFSCs or on 3D scaffolds [ 50 ] and Silk Fibroin Scaffolds [ 51 ] for APSC.…”

Comparative Proteomic and Metabolomic Analysis of Human Osteoblasts, Differentiated from Dental Pulp Stem Cells, Hinted Crucial Signaling Pathways Promoting Osteogenesis

“…Bone remodeling is a complex process controlled by multiple local and systemic factors. In order to study the mechanisms that regulate osteodifferentiation, DPSCs were grown in OsteoMAX-XF differentiation medium selected in our previous study [ 21 ]. The efficiency of differentiation was demonstrated (i) microscopically, (ii) spectrophotometrically, and (iii) immunocytochemically.…”

“…Various cell types (in particular MSCs) and multiple extracellular and intracellular signaling networks are involved in this well-organized physiological process [ 16 , 17 ]. Osteogenic microenvironment is often imitated in vitro by specific induction media [ 21 ]. The exact molecular mechanism by which cells differentiate and form a mineralized extracellular matrix is not yet fully understood.…”