Figure 6.

Blocking of autophagy inhibits wear particle induced osteoclastogenesis in vivo. The air pouch mold was established, and Ti wear particles with or without 3‐MA were injected in the air pouch. (a) After 7 days, the implanted bones were harvested to detect the mRNA expression of TRAP and Netrin‐1 with real‐time PCR. (b) The harvested bones were also subjected to immunohistochemical staining to detect the expression of TRAP and Netrin‐1. (c) The mRNA expression of RANKL and OPG in pouch wall tissues was detected by real‐time PCR. (d) The protein expression of RANKL and OPG in pouch wall tissues was detected by western blotting. (e) The serum levels of RANKL and OPG were detected by ELISA. The mRNA expression of proinflammatory factors (IL‐1β, IL‐6, and TNF‐α) in pouch wall tissues was measured to assess the degree of inflammation. Data represent mean ± SEM; n = 5 (*p < .05). ELISA, enzyme‐linked immunosorbent assay; IL, interleukin; mRNA, messenger RNA; TNF, tumor necrosis factor