Fig. 2

TOD inhibits RANKL-induced osteoclast differentiation and bone resorption without cytotoxicity in vitro. A The cell viability of BMMs exposed to TOD was measured by CCK-8. B The cell viability of RAW264.7 cells exposed to TOD was assessed by CCK-8. C BMMs were cultured with M-CSF (25 ng/ml), RANKL (50 ng/ml), and the indicated concentrations (0, 5, 10, and 20 µM) of TOD for 5 days. The effect of TOD on BMMs differentiation was detected using TRAP staining. Scale bar: 100 μm. D The area and number of osteoclasts were quantified per well. E BMMs were cultured with M-CSF (25 ng/ml) and RANKL (50 ng/ml) for 5 days, and TOD (20 µM) was added at different stages during osteoclast differentiation. The effect of TOD on BMMs differentiation was detected using TRAP staining. Scale bar: 100 μm. F The area and number of osteoclasts were quantified per well. G BMMs were cultured with M-CSF (25 ng/ml), RANKL (50 ng/ml), and the indicated concentrations (0, 5, 10, and 20 µM) of TOD for 5 days. Scanning electron microscopy was used to observe bone resorption pits. Scale bar: 100 μm. H The resorption area and number of rings were quantified. I BMMs were cultured with M-CSF (25 ng/ml) and RANKL (50 ng/ml) for 5 days, and TOD (20 µM) was added at different stages during osteoclast differentiation. Scanning electron microscopy was used to observe bone resorption pits. Scale bar: 100 μm. J The resorption area and number of rings were quantified. Data represent means ± SD of triplicate independent experiments. ^{*, #} indicates p < 0.05, ^**,## indicates p < 0.01, ^ns indicates not significant