Diseases caused by infection are a threat to human health and the world economy, with bacterial infections being responsible for a large portion of hospitalizations, morbidity, and mortality, which necessitates the quest for advanced medications and/or sustainable antibacterial strategies. This study aims to develop bioderived chitin nanofibers (ChNFs) and ZnO nanoparticles to produce non-toxic nanohybrid materials with improved aqueous stability and enhanced antibacterial properties. These nanohybrids were formed via either (i) an ex situ route by mixing the ChNFs with ZnO nanoparticles prepared by flame spray pyrolysis or (ii) an in situ route resulting in ZnO nanoparticles being formed and embedded into ChNFs by a simple aqueous hydrothermal process, utilizing a low-cost Zn inorganic precursor. The ChNFs, the ZnO nanoparticles, and the nanohybrids were physicochemically characterized for their size, morphology, charge and stability. Their antibacterial activity was evaluated against Gram (−) E. coli and Gram (+) S. aureus bacteria, while their cytocompatibility was assessed against mammalian cell lines. The obtained results reveal a balance between antibacterial activity and cytocompatibility, as both nanohybrids exhibited satisfactory antibacterial activity (MIC 200–300 μg/mL) combined with low cytotoxicity against mammalian cell lines (cell viability 80–100%), indicating that their further application as safe and effective antibacterial agents is promising.

The development of eco-friendly antimicrobial materials is essential for addressing antibiotic resistance, while reducing environmental impact. In this study, bio-derived anionic and cationic cellulose nanofibers (a-CNF and c-CNF) were employed as templating matrices for the in situ hydrothermal synthesis of cellulose/ZnO nanohybrids. Physicochemical characterization confirmed efficient cellulose functionalization and high-quality nanofibrillation, as well as the formation of uniformly dispersed ZnO nanoparticles (≈10–20 nm) strongly integrated within the cellulose network. The ZnO content was 30 and 20 wt. % for a-CNF/ZnO and c-CNF/ZnO, respectively. Antibacterial evaluation against Escherichia coli and Staphylococcus aureus revealed enhanced activity for both hybrids, with c-CNF/ZnO displaying the lowest MIC/MBC values (50/100 μg/mL). Antiviral assays revealed complete feline calicivirus inactivation at 100 μg/mL for c-CNF/ZnO, while moderate activity was observed against bovine coronavirus, highlighting the role of surface charge. Cytotoxicity assays on mammalian cells demonstrated high biocompatibility at antimicrobial concentrations. Life cycle assessment showed that c-CNF/ZnO exhibits a lower overall environmental burden than a-CNF/ZnO, with electricity demand being the main contributor, indicating clear opportunities for further reductions through process optimization and scale-up. Overall, these results demonstrate that CNF/ZnO nanohybrids effectively combine renewable biopolymers with ZnO antimicrobial functionality, offering a sustainable and safe platform for biomedical and environmental applications.