A water-soluble, blocked diisocyante was used to cross-link chitosan under various degrees of solvation, including hydration to form hydrogels. Thermal cross-linking of films cast from various amounts of organic cosolvents was found to increase with increased level of cosolvent up to a solvation level of 17% (w/w) and to be more efficient than for films prepared without cosolvent. Rheological studies revealed that gel modulus increased and gel time decreased with increasing cross-linker content and that gelation kinetics were consistent with a process having an activation energy of 103 kJ/mol. Swelling of hydrogels indicated that, even at high levels of hydration, the increased molecular mobility of reactants allowed for efficient network formation in a concentration-dependent manner. The extent of solvation via equilibrium swelling correlated well with degradative properties of chitosan networks in the presence of Chitinase (E. C. 3.2.1.14) from Streptomyces griseus with stability increasing with decreasing swelling (i.e., increased cross-linking).