This investigation aims to highlight the applicability of a potent eco-friendly developed composite film to combat the Escherichia coli biofilm formed in a model food system. ZnO nanoparticles (NPs) synthesized using green methods were anchored on the surface of cellulose nanocrystals (CNCs). Subsequently, nano-chitosan (NCh) solutions were used to disperse the synthesized nanoparticles and cinnamon essential oil (CEO). These solutions, containing various concentrations of CNC@ZnO NPs and CEO, were sequentially coated onto cellulosic papers to inhibit Escherichia coli biofilms on grey zucchini slices. Six films were developed, and Fourier transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, biodegradation, and mechanical properties were assessed. The film containing 5 % nano-emulsified CEO + 3 % dispersed CNC@ZnO nano-hybrid in an NCh solution was selected for further testing since it exhibited the largest zone of inhibition (34.32 mm) against E. coli and the highest anti-biofilm activity on biofilms developed on glass surfaces. The efficacy of the film against biofilms on zucchini surfaces was temperature-dependent. During 60 h, the selected film resulted in log reductions of approximately 4.5 logs, 2.85 logs, and 1.57 logs at 10 °C, 25 °C, and 37 °C, respectively. Applying the selected film onto zucchini surfaces containing biofilm structures leads to the disappearance of the distinctive three-dimensional biofilm framework. This innovative anti-biofilm film offers considerable potential in combatting biofilm issues on food surfaces. The film also preserved the sensory quality of zucchini evaluated for up to 60 days.