The role of the oral microbiome and its effect on dental diseases is gaining interest. Therefore, it has been sought to decrease the bacterial load to fight oral cavity diseases. In this study, composite materials based on chitosan, chitosan crosslinked with glutaraldehyde, chitosan with zinc oxide particles, and chitosan with copper nanoparticles were prepared in the form of thin films, to evaluate a new alternative with a more significant impact on the oral cavity bacteria. The chemical structures and physical properties of the films were characterized using by Fourier transform infrared spectroscopy (FTIR,) Raman spectroscopy, X-ray photoelectron spectroscopy (XPS), elemental analysis (EDX), thermogravimetric analysis (TGA), X-ray diffraction (XRD), scanning electron microscopy (SEM), and contact angle measurements. Subsequently, the antimicrobial activity of each material was evaluated by agar diffusion tests. No differences were found in the hydrophilicity of the films with the incorporation of ZnO or copper particles. Antimicrobial activity was found against S. aureus in the chitosan film crosslinked with glutaraldehyde, but not in the other compositions. In contrast antimicrobial activity against S. typhimurium was found in all films. Based on the data of present investigation, chitosan composite films could be an option for the control of microorganisms with potential applications in various fields, such as medical and food industry.