This study explores the green synthesis of zinc oxide (ZnO) nanoparticles (NPs) using Aloe vera leaf extract (ALE) as both the reducing and capping agents. The ZnO NPs were characterized using UV, FTIR, HR-TEM, and XRD, revealing a pure crystalline state with an average crystallite size of approximately 25 nm. A regenerated cellulose film (RCF) incorporating these ZnO NPs was prepared through the immersion phase-conversion method. The structure and morphology of the ZnO/RCF were characterized using SEM and EDS mapping, showing well-dispersed and stabilized ZnO NPs in the cellulose matrix. The antimicrobial properties of the ZnO/RCF were evaluated against *Kosakonia Cowanii* and *Alternaria alternata* strains, with the highest inhibition zone observed for ZnO-10/RCF (22 ± 0.4 mm). Additionally, the ZnO/RCF exhibited antioxidant activity as a DPPH free radical scavenger. The study suggests that ZnO/RCF has potential applications in food packaging, pharmaceuticals, agriculture, and cosmetics.This study explores the green synthesis of zinc oxide (ZnO) nanoparticles (NPs) using Aloe vera leaf extract (ALE) as both the reducing and capping agents. The ZnO NPs were characterized using UV, FTIR, HR-TEM, and XRD, revealing a pure crystalline state with an average crystallite size of approximately 25 nm. A regenerated cellulose film (RCF) incorporating these ZnO NPs was prepared through the immersion phase-conversion method. The structure and morphology of the ZnO/RCF were characterized using SEM and EDS mapping, showing well-dispersed and stabilized ZnO NPs in the cellulose matrix. The antimicrobial properties of the ZnO/RCF were evaluated against *Kosakonia Cowanii* and *Alternaria alternata* strains, with the highest inhibition zone observed for ZnO-10/RCF (22 ± 0.4 mm). Additionally, the ZnO/RCF exhibited antioxidant activity as a DPPH free radical scavenger. The study suggests that ZnO/RCF has potential applications in food packaging, pharmaceuticals, agriculture, and cosmetics.