Nox2 Activity Is Required in Obesity-Mediated Alteration of Bone Remodeling

Figure 4

Effect of Nox2 on high-fat-diet- (HFD-) induced changes in bone marrow osteoclastogenesis. Eight-week-old WT and Nox2-KO mice were fed with an SD or an HFD for 3 months. (a) Bone marrow cells were isolated from the left femur and tibia of each group. Bone marrow cells (1 × 10⁶) were cultured in the presence of RANKL and MCSF for 5 days. Cells were then stained for TRAP and counted under a microscope. TRAP-positive multinucleated cells (MNCs) containing more than 3 nuclei were considered osteoclasts. Bars show the number of TRAP-positive MNCs in each group expressed as a number of cells per field. (b) Right femurs were crushed under liquid nitrogen, and RNA was isolated and analyzed for mRNA levels for TNF-α, IL-6, RANKL, and cathepsin K (CTSK) by qRT-PCR. mRNA Ct values for these genes were normalized to the housekeeping gene 18S. Data are expressed as mean ± SEM (, 5 mice/group). Two-way ANOVA followed by post hoc Tukey’s multiple comparison test was used to assess differences among groups with significance defined as . ^§Significantly different from WT-SD. ^#Significantly different from WT-HFD. Significantly different from SD-KO.

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