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Zinc oxide (ZnO) is an inorganic compound widely used in everyday applications. ZnO is currently listed as a generally recognized as safe (GRAS) material by the Food and Drug Administration and is used as food additive. The advent of nanotechnology has led the development of materials with new properties for use as antimicrobial agents. Thus, ZnO in nanoscale has shown antimicrobial properties and potential applications in food preservation. ZnO nanoparticles have been incorporated in polymeric matrices in order to provide antimicrobial activity to the packaging material and improve packaging properties. This review presents the main synthesis methods of ZnO nanoparticles, principal characteristics and mechanisms of antimicrobial action as well as the effect of their incorporation in polymeric matrices. Safety issues such as exposure routes and migration studies are also discussed.