Figure 6.

Skeletal stem cell (SSC) viability, proliferation, and functionality over time. (A) Merged Calcein AM, Ethidium homodimer-1, and Brightfield images of SSCs on ethylene oxide sterilization (EOS) templates at days 1 and 14 demonstrated good cell viability over time, with a concomitant increase in cell density and stellate morphology. Dotted white circles indicate the area visualized under 10× magnification. Yellow scale bar represents 500 µm, and white scale bar represents 200 µm. (B) WST-1 proliferation assay demonstrated a higher proliferative rate of SSCs on TCP when compared to EOS templates over the 21 day culture period (p < 0.001). SSCs cultured in osteogenic media had a lower proliferative rate compared to SSCs cultured in basal media between days 7 and 21 (p < 0.001), reflecting osteogenic differentiation under 1,25-VitD₃ induction. (C) Alkaline phosphatase (ALP) activity peaked at day 7 in all three groups, with an expected stepwise reduction over the following 2 weeks of culture.