Nanocellulose/Fe₃O₄/Ag Nanozyme with Robust Peroxidase Activity for Enhanced Antibacterial and Wound Healing...

Peroxidase memetic nanozymes with their free radical-mediated catalytic actions proved as efficacious antibacterial agents for combating bacterial resistance. Herein, nanocellulose (NC) extracted from Eragrostis teff straw was used to prepare NC/Fe₃O₄/Ag peroxidase nanozyme as an antibacterial and wound healing agent. Characterization of the nanozyme with XRD, FTIR, SEM-EDX, and XPS confirmed the presence of silver NPs and the magnetite phase of iron oxide dispersed on nanocellulose. The peroxidase activity of the prepared nanozyme was examined using TMB and H₂O₂ as substrates which turned blue in acidic pH (λ_max = 652 nm). With a lower K_m (0.387 mM), the nanozyme showed a comparable affinity for TMB with that reported for the HRP enzyme. Furthermore, the nanozyme remained efficient over a broader temperature range while maintaining 61.53% of its activity after the fourth cycle. In vitro, antibacterial tests against Escherichia coli (Gram-negative) and Staphylococcus aureus (Gram-positive) bacterial strains showed that NC/Fe₃O₄/Ag exhibits concentration-dependent and enhanced antibacterial effect for Escherichia coli compared to NC and NC-Fe₃O₄ and negative control. Furthermore, the wound-healing performance of the NC-Fe₃O₄-Ag nanozyme was investigated in vivo using an animal model (mice). The nanozyme showed 30% higher wound healing performance compared to the control base ointment and is comparable with the commercial nitrofurazone ointment. The results show the potential of the prepared nanozyme for wound-healing purposes.