FIGURE 4.

CoNPs‐induced osteoblasts ferroptosis in vitro. (A) Effect of gradient concentrations (0, 25, 50, 100, 200, 400 μg/ml) of CoNPs on the osteoblast survival was observed by CCK‐8. (B) Transmission electron microscopy of osteoblasts after treatment with gradient concentrations of CoNPs for 24 h. Scale bar: 20 μm (upper) and 2 μm (lower). (C) Western blots performed after MC3T3‐E1 cells were treated with CoNPs for 24 h (n = 3). (D) ImageJ software was used to quantify the density of the western blot bands shown in (C). (E) Relative mRNA expression of Gpx4, Slc7a11, Acsl4, and Ptgs2 in each group. (F) ROS generation was detected by flow cytometry with DCFH‐DA (10 mM) in each group. (G) Lipid peroxide MDA level in each group. (H) The GSH level in each group. (I) Cell lipid peroxidation was detected by BODIPY 581/591 C11 staining using flow cytometry in each group. ^{*, #} indicates p < 0.05, ^{**, ##} indicates p < 0.01, ^ns indicates not significant