Hexabromocyclododecane (HBCD) induced PANoptosis of chondrocytes via activation of the NLRP3 inflammasome and decreased the exercise ability of mice in vivo

Hexabromocyclododecane (HBCD) is a persistent organic pollutant (POP). HBCD is found in the blood and tissues of most populations and causes a range of toxicological damage to tissues and cells. However, the toxicological effects of HBCD on chondrocytes are not fully understood. Here, we evaluated the toxicological effects of HBCD on chondrocytes and cartilaginous tissue. For this, a model of primary cartilage cells was established. Chondrocytes were exposed to different concentrations of HBCD. Western blot, indirect immunofluorescence, ELISA and other biochemical experiments were performed to analyze the toxicological effects of HBCD on chondrocytes/articular cartilage tissue. Cell proliferation assays showed that HBCD caused a reduction in the proliferative capacity of chondrocytes, and further work indicated that HBCD induces chondrocyte death. Further experiments demonstrated that HBCD caused an inflammatory response in chondrocytes by evaluating the levels of inflammatory factors. We found that HBCD led to PANoptosis in chondrocytes by detecting panapoptosis-related marker molecules, and experimental data indicated that apoptosis markers (cleaved caspase-3/7), pyroptosis markers (caspase-1/GSDMD-N) and necroptosis markers (pMLKL/RIPK3) were upregulated after HBCD treatment. Subsequent experiments illustrated that HBCD activated the DAMP sensor NLRP3, which then mediated ZBP1-induced PANoptosis. In the in vivo model, the experimental animals were administered HBCD at 25, 50 and 100 µg/kg/week for 15 weeks. We found that HBCD led to an inflammatory response in articular cartilage tissue. The safranin O-fast green assay showed a certain degree of damage to cartilage tissue under HBCD treatment. Furthermore, HBCD resulted in an increase in MMP13 expression and a downregulation of COL2 expression in chondrocytes/cartilaginous tissues. HBCD decreased the exercise ability of mice in vivo. These data indicate that HBCD leads to chondrocyte damage. In summary, this study lays the foundation for further exploration of the toxicological effects of HBCD on bone and joints.