The aim of this study was the preparation and characterization of chitosan sponges including a model antibiotic (i.e., norfloxacin). The chitosan sponges were prepared by a solvent evaporation method. The matrix was also cross-linked during the preparation. The results indicated that the chitosan sponges were in the fibrillar structure. The swelling behavior, norfloxacin loading, in vitro release characteristics, and antibacterial activity were determined. The effects of cross-linker concentration, norfloxacin/chitosan ratio, chitosan molecular weight, and base concentration were investigated. The most effective parameter was found to be the degree of neutralization. It was also observed that the equilibrium swelling ratio decreased with increasing cross-linking density. The norfloxacin release was found to be swelling controlled initially and diffusion controlled at the extended release periods. It was also found that the antibacterial activity was directly proportional to the release rate.