Figure 4.

Melatonin restores oxidative stress-inhibited osteoblast differentiation of human MSCs by activating AMPK signaling. (A) Human MSCs were treated with 100 μM H₂O₂ and incubated in ODM with the indicated concentrations of melatonin for 21 days. The calcium deposition of human MSCs was assessed by Alizarin Red S staining. (B) The mineralized layers were dissolved and quantified using a microplate reader at 570 nm. (C) Total proteins were subjected to Western blot analysis using the specified antibodies. Data are represented as mean ± SEM of three individual experiments (n = 3). Statistical significance was determined using Student's t-test (^*p < 0.05 and ^**p < 0.01 in contrast to the group treated with H₂O₂ alone).