BACKGROUND: Pain reduction, immunomodulation, and cartilage repair are key therapeutic goals in osteoarthritis (OA) treatment. In this study, we evaluated the therapeutic effects of porcine cartilage acellularized matrix (pCAM) derived from naive tissue and compared it with the synthetic material polynucleotides (PN) for OA treatment. METHODS: pCAM was produced from porcine cartilage through physicochemical processing. LC-MS protein profiling identified the key proteins. In vitro experiments involved treating human synovial cell with pCAM and PN to assess cell viability and reductions in pro-inflammatory cytokines (IL-1β and IL-6). In vivo studies utilized a rat DMM-induced OA model. Pain was evaluated in weight-bearing tests, and inflammation reduction was confirmed using specific macrophage markers of CD68, CD86, and CD163 in immunohistochemical staining of synovial tissue. Cartilage regeneration was evaluated by histopathological analyses. RESULTS: The major protein components of pCAM include factors integral to cartilage and ECM integrity. They also contain proteins that help reduce inflammation. In vitro studies revealed a decrease in pro-inflammatory cytokines and survival of synovial cells were observed. In vivo treatment with pCAM resulted in a reduction of pain and inflammation, while promoting cartilage regeneration, thereby accelerating the healing process in OA. CONCLUSION: Our findings suggest that pCAM may contribute to the treatment of OA by alleviating synovial inflammation and supporting cartilage regeneration, thereby addressing both the inflammatory and degenerative aspects of the disease.