Fig. 4

Y-27632 and paclitaxel impact osteogenic/adipogenic gene expression. a Gene expression analyses for markers of adipogenesis (PPARγ and C/EPBα) and osteogenesis (RUNX2 and BGLAP) in hMSC cultured for 72 h with basal culture medium (−Y) or treated with 10 μM Y-27632 (+Y, n ≥ 8). Expression levels (mean + s.d.) are shown as fold change normalized to expression in undifferentiated hMSC (set to 1). b Representative micrograph of hMSC (5 × 10⁶ cells mL⁻¹) encapsulated in a 1:0.75 hydrogel after 28 days showing fibronectin, α₅ integrin, and DAPI. Fibronectin co-localized with punctate α₅ integrin by Manders’ coefficients: Manders’ tM1 = 0.83 ± 0.18 and Manders’ tM2 = 0.84 ± 0.10 (n = 4, 25 cells). c Representative micrographs of hMSC within hydrogels after 72 h and stained with Phalloidin-TRITC (actin) and DAPI. d Gene expression analyses for markers of adipogenesis (PPARγ and C/EPBα) and osteogenesis (RUNX2 and BGLAP) in hMSC cultured for 72 h with basal culture medium (−PTX) or treated with 50 nM paclitaxel (+PTX, n ≥ 8). Expression levels (mean + s.d.) are shown as fold change normalized to expression in undifferentiated hMSC (set to 1). e Representative micrographs of hMSC (5 × 10⁶ cells mL⁻¹) within a 1:0.75 hydrogel for 72 h and stained for tubulin and DAPI. Scale bars in b, c, and e are 100 µm, and in insets = 10 µm. In a and d a Mann–Whitney test (two-tailed) was used to detect statistical significance, *p < 0.05, **p < 0.01, ***p < 0.001