The objective of this study was to synthesize and evaluate the biological effectiveness of cerium oxide (CeO2) against various bacterial and fungal strains. The chitosan‐CeO2 nanocomposite was comprehensively characterized. The Fourier transform infrared (FTIR) spectra displayed a combination of bands that provided evidence for the existence of a composite complex between the chitosan (CS) structure and the Ce–O bands originating from CeO2 nanoparticles. SEM measurements showed uniform dispersion across the whole CS surface, and the morphological changes were clearly observed as the CeO2 nanoparticle content was increased. In contrast, CS‐CeO2 demonstrates enhanced antibacterial efficacy against both Gram‐positive and Gram‐negative pathogens. The MTT method was utilized to assess the cytotoxicity of the examined CS‐CeO2, with the aim of evaluating the cell viability of various cancer cell lines under varying ultraviolet (UV) exposures. The enhanced penetration of CS and CeO2 nanocomposites into cancer cells during their anticancer activity, even in the absence of UV light, led to an increased rate of cell death. The administration of greater dosages of CS and its CeO2 nanocomposites led to an increased generation of reactive oxygen species (ROS), which was observed to be associated with an enhancement in the efficacy of anticancer activity. The impact was additionally amplified through exposure to UV radiation. The nanocomposite thin films demonstrated much higher antioxidant activity against the DPPH radical as compared to ordinary vitamin C. The aforementioned observation was derived from an evaluation of the antioxidant properties of the subject utilizing the DPPH free radical scavenging technique.