<p>Zinc oxide nanoparticles (ZnONPs) have garnered significant attention in the biomedical field due to their unique properties and versatile applications. This review provides a comprehensive overview of various synthesis methods for ZnONPs, including chemical, physical, and green synthesis approaches, which yield nanoparticles of diverse sizes, shapes, and functional characteristics. Characterization techniques are crucial for understanding the structural and morphological properties of ZnONPs, facilitating the development of innovative designs for biomedical applications. The review highlights recent advancements in the use of ZnONPs for drug delivery systems, tissue regeneration, bioimaging, and cancer treatment protocols. Additionally, their notable antimicrobial activities against a range of pathogens position ZnONPs as promising candidates for infection control and wound healing therapies. Although ZnONPs have great potential, safety concerns must be addressed, and their properties should be optimized to reduce risks. Future research should focus on establishing industry, governmental, and academic consensus on safety protocols to ensure public health protection while maximizing the benefits of ZnONPs in various biomedical domains. This review aims to provide insights into the multifaceted applications of ZnONPs and their role in advancing medical science. </p> Graphical Abstract <p></p>