Ferroptosis-related genes and pathways in knee osteoarthritis cartilage degeneration: based on bioinformatics technology and vivo experimental verification

Objective To thoroughly explore the function and control networks of genes related to ferroptosis in the breakdown of knee osteoarthritis (OA) cartilage, and to identify potential therapeutic targets for the disease. Methods This study combined comprehensive bioinformatics analysis with in vitro and in vivo experiments. Weighted gene co-expression network analysis (WGCNA) was used to analyze gene expression data sourced from the Gene Expression Omnibus (GEO) database.differential gene expression screening, and pathway enrichment analysis. An OA mouse model was established through surgical induction, and pathological and biochemical changes were assessed using micro-CT, ELISA, histological staining, and immunohistochemistry. Clinical cartilage samples from OA patients and controls were also analyzed using immunofluorescence, Western blot, and real-time PCR. Results Bioinformatics analysis identified 62 ferroptosis-related genes, with key genes including MYC, MMP2, SPP1, and BNIP3. Pathway analysis revealed significant enrichment of the ferroptosis pathway, mTOR, and HIF-1 signaling pathways. In the OA mouse model, significant pathological changes in subchondral bone and articular cartilage were observed, including increased iron deposition, reduced mitochondrial cristae, and upregulation of oxidative stress markers. Immunohistochemical and immunofluorescence analyses showed that the negative regulator GPX4 was downregulated, while the positive regulator p53 was upregulated in the OA group. Western blot and RT-PCR confirmed the differential expression of key ferroptosis-related genes in both animal and clinical samples. Conclusion Our findings indicate that ferroptosis may promote the development of OA by influencing iron metabolism, oxidative stress responses, and cartilage matrix degradation. Key ferroptosis-related genes, such as MYC, MMP2, SPP1, and BNIP3, and their associated pathways, were identified as potential therapeutic targets.