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The knowledge of agarose (AG) oxidation using periodate as oxidizer has not been systematically explored. This paper synthesized oxidized agarose (OAG) using solid-sate and solution reaction methods; the reaction mechanism and the properties of OAG samples were systematically evaluated. Chemical structure analysis disclosed that the aldehyde group and carboxyl group contents in all OAG samples are extremely low. Meanwhile, crystallinity, dynamic viscosity and molecular weight of OAG samples is lower than that of the original AG. Reaction temperature, time and sodium periodate dosage are inversely proportional to the decline of the gelling temperature (Tg) and melting temperature (Tm); and the Tg and Tm for the OAG sample obtained are even 19 °C and 22 °C lower than that of the original AG. The as-synthesized OAG samples all possess excellent cytocompatibility and blood compatibility; and can promote the proliferation and migration of fibroblast cells. Last but not least, the gel strength, hardness, cohesiveness, springiness and chewiness of the OAG gel can be effectively regulated via oxidation reaction. In conclusions, both solid and solution oxidation can regulate the physical properties of OAG and enlarge its potential applications in wound dressing, tissue engineering and food areas., Competing Interests: Declaration of competing interest The authors report no declarations of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)