BACKGROUND: Osteoarthritis (OA) is the most prevalent joint degenerative disease. MF is considered as a first-line treatment for OA. In the long term, the cartilage tissue regenerated after MF is fibrocartilage. In this study, we examine whether combined treatment of MF and Platelet lysate (PL) can inhibit promotion of cartilage repair and antifibrosis. METHODS: OA rat model established by the modified Hulth method. Rat PL injected into treated knee joints after MF surgery. The expression levels of metabolic and fibrosis molecules (Col2, Mmp13, Col1, Col3, α-SMA, and Ctgf) of chondrocytes were examined by immunohistochemistry. Cell immunofluorescence was used to assess bone marrow MSCs (BMSCs) proliferation. Transwell assays evaluated BMSCs migration, and qPCR and Western blot analyzed the mechanisms of PL. Moreover, a retrospective analysis was conducted to determine the clinical efficacy and safety of the combined treatment of MF and PL on OA patients. RESULTS: In vivo data showed that the combined treatment of MF and PL significantly alleviated joint pain, protected chondrocytes and inhibited synovial fibrosis on OA rats, as was confirmed by upregulation of Collagen II and downregulation of Mmp13, Col1, Col3, α-SMA, and Ctgf. Such anti-OA and antifibrosis effects of the combined treatment of MF and PL were superior to MF alone. In vitro data showed that PL induced cellular chondrogenic differentiation and migration of BMSCs, suggesting that PL facilitated stem cell homing to the cartilage injury sites and promoted cartilage repair and regeneration. Furthermore, the clinical data showed significant improvements of pain reduction and cartilage repair in OA patients. CONCLUSION: This study demonstrated the anti-OA and antifibrosis effects of the combination of MF and PL, providing a promising synergistic therapeutic option for the treatment of OA.