Chitosan is a potential material for biomedical applications. Recently, it is widely applied in bone tissue regeneration, drug delivery systems and wound dressings. Chitosan has outstanding biological properties, including biocompatible, biodegradable and antibacterial ability. Moreover, chitosan has hemostatic ability for promoting blood to coagulate together. However, the applications of chitosan are limited for its low mechanical properties and rapid degradation trend, especially in the environment containing lysozyme. Chitosan existing in acid environment becomes a cationic polyelectrolyte due to its amine group, it can easily form polyelectrolyte complexes with other anionic polyelectrolytes and growth factors, or form crosslink structure with carboxylic group by composing amide bond. Through this strategy, the properties of chitosan can be modified and enhanced. In this research, in order to enhance the mechanical properties and to ease the biodegradation rate, the glutamic acid, succinic acid and succinic anhydride are applied to form crosslink structure with chitosan separately, with the repeating unit ratio of 80:20 to 50:50. Besides, this study investigates physicochemical characterization and biological properties of the crosslinked hydrogels. Via the implant study, animal test was conducted to evaluate the possibility that crosslinked hydrogels could be a potential wound dressing material. Results of wound closure and histological tissue observation showed that chitosan/diacid crosslinked hydrogels could accelerate the process of wound healing and tissue regeneration. The study reveals chitosan could suppress the infiltration of inflammatory cells and accelerate fibroblast proliferation while the diaicds could enhance epithelial migration. Thus, chitosan/diacid crosslinked hydrogels could be potential materials for wound dressing applications.