Fig. 1: Inhibition of osteoclastogenesis by ASMSCs compared with HDMSCs.

CD14 + monocytes were cultured with HDMSCs or ASMSCs in the presence of M-CSF and RANKL. a Representative images of TRAP staining of osteoclasts co-cultured at different time points (× 100). b The number of TRAP⁺ osteoclasts in each well from cultures at different time points is shown. c Representative images of osteoclasts stained with FITC-phalloidin at different time points (× 200). d Representative images for bone resorption assays at different time points ( × 200). Cells cultured with HDMSCs or ASMSCs on bovine cortical slides were stained with toluidine blue. e Pit formation on each slide was assessed. f mRNA expression levels of TRAP, CTSK, and NFATc1 in osteoclasts were determined by qPCR on day 9. g Protein levels of TRAP, CTSK, and NFATc1 in osteoclasts were determined by western blot analyses on day 9. h Quantitative data of TRAP, CTSK, and NFATc1 protein levels determined by western blot analyses are shown. i Activation of signaling pathways involved in osteoclastogenesis was determined by western blot analyses on day 9. j Quantitative data for activation of signaling pathways determined by western blot analyses are shown. Values are the mean ± SD of 30 samples per group. The results represent three independent experiments. *, p < 0.05; HDMSCs, mesenchymal stem cells from healthy donors; ASMSCs, mesenchymal stem cells from patients with ankylosing spondylitis