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BACKGROUND: During healing process of chronic wounds, excessive production of reactive oxygen species can impair the function of L929 fibroblasts, thereby delaying wound repair. Therefore, protecting fibroblasts from oxidative stress is important to promote wound healing. OBJECTIVE: To assess the protective effects of carboxymethyl chitosan-oxidized chondroitin sulfate/platelet-rich plasma (CMC-OCS/PRP) hydrogel on L929 cells under H2O2 stimulation. METHODS: CMC-OCS/PRP hydrogels were prepared, and the micromorphology, degradation performance, scavenging ability of H2O2 and hydroxyl radical and biocompatibility of the hydrogels were characterized. L929 cells with good growth state were taken and cultured in five groups. The control group was cultured conventionally. H2O2 was added to the H2O2 group. Carboxymethyl chitosan-oxidized chondroitin sulfate hydrogel extract + H2O2 was added to the CMC- OCS group. Platelet-rich plasma gel extract + H2O2 was added to the PRP group. The CMC-OCS/PRP group was treated with carboxymethyl chitosan-oxidized chondroitin sulfate/platelet-rich plasma hydrogel extract + H2O2. Each group was treated with hydrogel extract for 6 hours, and then H2O2 for 24 hours. After culture, the levels of active oxygen and malondialdehyde, apoptosis and expression of collagen fiber I protein were detected. In the presence of H2O2, the above hydrogel extracts were directly or indirectly co-cultured with L929 fibroblasts for 36 hours, respectively. Migration ability of the cells was detected by scratch test and Transwell chamber test. RESULTS AND CONCLUSION: (1) CMC-OCS/PRP hydrogels had uniform and interrelated porous structure and good degradation ability, could effectively remove H2O2 and hydroxyl radicals in vitro, and had good biocompatibility. (2) Compared with the control group, the apoptosis rate, reactive oxygen species, and malondialdehyde levels were increased (P < 0.05); the spread area of cells was decreased (P < 0.05), and the expression of collagen fiber I protein had no significant changes (P > 0.05) in the H2O2 group. Compared with the H2O2 group, reactive oxygen species level was decreased in the CMC-OCS group (P < 0.05), malondialdehyde level was decreased (P < 0.05), and cell spread area was increased (P <0.05) in the PRP group, CMC-OCS group, and CMC-OCS/PRP group; apoptosis rate was decreased in the CMC-OCS/PRP group (P < 0.05), and collagen fiber I protein expression was increased in the PRP group, CMC-OCS group, and CMC-OCS/PRP group (P < 0.05). (3) Compared with the control group, the number of cell migration was decreased (P < 0.05), and the migration area had no significant change (P > 0.05) in the H2O2 group. Compared with the H2O2 group, the number and area of cell migration were increased in the PRP group, CMC-OCS group, and CMC-OCS/PRP group (P < 0.05), and the increase was most significant in the CMC-OCS/PRP group. (4) Under oxidative stress, CMC-OCS/ PRP hydrogel can improve the migration ability of fibroblasts, resist cell apoptosis, and preserve cell extension function [ABSTRACT FROM AUTHOR]