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Objective: The objective of this study was to prepare a controlled release system, which could be used in medical applications in order to treat infections at the host region. Methods: Microlayer films were prepared via solvent casting method. Films with 3-layers were prepared by using chitosan (CHI), gelatin (GEL) and alginate (ALG) in the form of CHI /ALG/CHI and CHI-GEL /ALG/CHI-GEL with or without ceftriaxone sodium (CS) which is loaded in the middle ALG layer. All films were crosslinked by exposing them to glutaraldehyde (GA) vapor for different (2 h, 10 h or 24 h) durations. Mechanical properties of the films and release kinetics of CS at three different pH conditions (pH 5.5, 7.4, and 10.0) were investigated. The antibacterial efficiency of the released CS against Escherichia coli was examined via agar spot test. Results: The results indicated that the presence of GEL in the upper and lower layers of the 3-layer construct prevented fragility and increased the mechanical strength of the films, whereas the presence of CS in the middle layer caused decrease in the mechanical properties. Crosslinking with GA did not demonstrate a significant effect on the release profile of CS, but due the interaction of chitosan with alginate and CS, the drug release was delayed. Antibacterial tests which were carried out by using the released media of drug loaded films displayed inhibition zone in agar spot tests against E. coli. Conclusion: It was concluded that multilayered films prepared by solvent casting can be good candidates as drug carrier devices in medical applications. [ABSTRACT FROM AUTHOR]