Chitosan is the most important derivative of chitin, the second most abundant biopolymer just after cellulose, which has received great attention because of its unique characteristics. Without doubt, its biomedical usages have gained more importance among the vast variety of chitosan applications owing to its good biocompatibility and biodegradability. In recent years, particular interest has been devoted to chitosan hydrogels as a promising alternative in competition with conventional sutures or bioadhesives. In the current work, we have investigated the effectiveness of chitosan hydrogel to stop bleeding and air leakaging of lung fistula. Polycationic chitosan was obtained with solubilization of chitosan powder in aqueous acidic media. Different parameters such as acid type and concentration, and degree of deacetylation (DD%) of chitosan, were altered to modify hydrogel properties including viscosity, pH, cohesive strength, and tissue bioadhesiveness. In vivo experiments have been conducted on sheep models which provide a convenient way to evaluate the efficacy of prepared samples. The lung was punctured in distinctive geometries and hydrogel then injected on. Bioadhesive strength as well as irritant effects were discussed. Samples with higher degree of deacetylation, includig Chs-16 (DD%=99, MW=230,000) and Chs-19 (DD% = 98, MW=300,000) that were dissolved in lactic media showed best sealing effect. Further studies are now conducted to optimize the sealing properties of chitosan based hydrogels.