Comparative evaluation of biologically and chemically synthesized zinc oxide nanoparticles for preventing Candida auris biofilm.

Candidozyma auris (formerly Candida auris) is a multidrug-resistant yeast that poses a significant global health threat due to its ability to form biofilms and resist various antifungal treatments. This study evaluates and compares the antifungal efficacy of biologically synthesized zinc oxide nanoparticles (ZnO-NP-B) and chemically synthesized ZnO nanoparticles (ZnO-NP-C1 and ZnO-NP-C2), developed using the dry-wet chemical method and sol-gel method, respectively. ZnO-NP-B was synthesized using Lactobacillus gasseri. The nanoparticles were characterized for size, charge, and morphology using Particle Size Analyzer, photon correlation spectroscopy with a 90 Plus Zetasizer, and scanning electron microscopy (SEM), respectively. The antifungal activity was assessed through minimum inhibitory concentration (MIC) determination, biofilm inhibition assays by XTT assay, and gene expression analysis. ZnO-NP-C1 exhibited the highest antifungal activity against C. auris planktonic cells, with a MIC value of 61.9 ± 3.3 µg/ml, followed by ZnO-NP-C2 (151 ± 7.83 µg/ml), whereas ZnO-NP-B showed limited efficacy (MIC = 1 mg/ml). Chemically synthesized ZnO-NPs, particularly ZnO-NP-C2, did not induce overexpression of resistance genes (CDR1, MDR1, ERG2, ERG11, FKS1, CHS1), whereas ZnO-NP-B triggered their upregulation, potentially promoting resistance. ZnO-NP-C1 was the most effective in preventing biofilm formation, reducing C. auris adhesion by 67.9 ± 2.35% at 150 µg/ml, while ZnO-NP-B exhibited negligible inhibition. Gene expression analysis further confirmed that ZnO-NP-C1 significantly downregulated adhesive genes (ALS5, IFF4, CSA1) by up to 0.37 ± 0.006, 0.043 ± 0.002, and 0.06 ± 0.0004, respectively. These findings highlight the potential of ZnO-NP-C1 as a promising antifungal agent for preventing C. auris biofilms, emphasizing the critical role of synthesis methods in optimizing nanoparticle properties for antifungal applications.