Biodegradable chitosan films were produced using lactic and acetic acid solutions and glutaraldehyde as crosslinking agent by casting technique. The aim of this study was to evaluate the influence of different acids and the crosslinking on the characteristics and biodegradability of chitosan films. The films were analyzed through thickness, swelling degree, mechanical and thermal properties, chemical structure and biodegradability. The glutaraldehyde release was also evaluated. All films' properties are deeply affected by the acid type and glutaraldehyde crosslinking. Crosslinking reduced the swelling degree of films, also increasing their fragility. The glutaraldehyde release from the films to the simulant solutions was not observed after the incubation period. Through biodegradation tests, it was observed that the crosslinking does not prevent films' degradability but longer time is required. The qualitative analysis of ecotoxicity of the films suggests the possibility of composting the developed films. The films presented potential for application as membranes and in packaging, and different formulations can be used according to the desired final characteristics.