Fig. 5.

Osteogenic differentiation effects of the RSF-based hydrogel samples on the BMSCs and their immunomodulatory effect on macrophage polarization in RAW264.7 cells with or without ES. (A, B) Representative photographs of ALP staining on day 7 and ARS staining on day 21. (C–F) Representative immunofluorescence staining and quantitative analysis of Runx-2 and Col-1 proteins after 7 days. (G) Real-time PCR results for the mRNA expression of OCN, Runx-2, Col-1, and ALP on day 7 and 14. (H) Flow cytometry results of the percentage of CD206⁺F4/80⁺cells treated with different interventions for 48 h. (I) Representative immunofluorescence staining of Arginase-1. (J) Schematic diagram showing the process of osteogenic differentiation of BMSCs regulated by macrophage M2 polarization. (K) Quantitative analysis of ALP staining of BMSCs after being cultured using the conditioned medium of RAW264.7 cells for 7 days. **P < 0.01, *P < 0.05, n = 3.