Figure 8.

Osteogenic differentiation of DPSCs encapsulated in hydrogel (3D) or seeded on the surface coated with gelatin (2D) cultured in hypoxic (2% O₂) or normoxic (about 18% O₂) environment. (a) Quantitative analysis of mRNA expression for osteogenesis associated genes (Col1A, Runx2) in DPSCs on days 7, 14, and 21 of differentiation. Fold change in the expression of analysed genes in control cells before differentiation was calculated as 1.0 and marked by a solid line. Results are presented as mean ± SEM, n = 3 (every sample was analysed in duplicate). p < 0.05 (t-test). (b) Representative images of DPSCs differentiated into osteoblasts on days 7, 14, and 21 days of differentiation. Panel “Background” contains the representative images of 2D and 3D surfaces (standards plastic dish and hydrogel without cells, respectively) stained with Alizarin Red S solution to visualize background staining (on day 7 post surface preparation). Panels “Day 7”, “Day 14”, “Day 21” demonstrate representative images of DPSCs cultured in StemPro osteogenesis differentiation kit. On days 7, 14 and 21, DPSCs were fixed with paraformaldehyde and stained with Alizarin Red S (red staining of deposits of calcium phosphate is a characteristic for osteogenic differentiation).