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<p><i>Pseudomonas aeruginosa</i> is a highly adaptable opportunistic pathogen frequently associated with chronic and hard-to-treat infections, particularly in burn units and immunocompromised patients. Its intrinsic and acquired resistance to multiple antibiotics poses a major therapeutic challenge. While ZnO nanoparticles conjugated with thiosemicarbazone (TSC) have shown promise in general antimicrobial applications, their potential for simultaneously inhibiting biofilm formation and pyocyanin production—key virulence factors—in clinical <i>P. aeruginosa</i> strains remains unexplored. In this study, ZnO nanoparticles were synthesized via a hydrothermal route and conjugated with a glutamine-modified TSC ligand (ZnO@Glu-TSC) to enhance their antimicrobial efficacy. The nanoconjugate was comprehensively characterized using UV–Vis spectroscopy, X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDX). Functional evaluations were conducted against clinical isolates of <i>P. aeruginosa</i>, including minimum inhibitory concentration (MIC), fractional inhibitory concentration (FIC) index, biofilm inhibition, and pyocyanin suppression assays. ZnO@Glu-TSC nanoparticles exhibited a sharp UV–Vis absorption peak at 380 nm with a band gap of 3.26 eV, and XRD confirmed a hexagonal wurtzite structure with an average crystallite size of ~ 19.8 nm. The nanoconjugate demonstrated significantly enhanced antibacterial activity with MIC values ranging from 128 to 512 µg/mL and synergistic effects in 70% of clinical isolates (FIC ≤ 0.5, <i>p</i> &lt; 0.01). Biofilm inhibition assays revealed an 80% reduction in biomass (OD values approaching those of the negative control), while pyocyanin production decreased by more than 75% at 512 µg/mL (<i>p</i> &lt; 0.001). These results represent the first demonstration of ZnO@Glu-TSC's dual antivirulence action against clinical <i>P. aeruginosa</i> strains, underscoring its therapeutic promise as a potent, multi-targeted nanoantimicrobial candidate and warranting further development for translational nanomedicine applications in combating persistent infections.</p>

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Novel dual-targeting of biofilm formation and pyocyanin production in clinical Pseudomonas aeruginosa isolates using glutamine-modified thiosemicarbazone-conjugated ZnO nanoparticles

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年11月10日

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  • Hananeh Mahboob Rabani,
  • Khosro Isazadeh,
  • Mohammad Faezi Ghasemi,
  • Alireza Habibi

摘要

Pseudomonas aeruginosa is a highly adaptable opportunistic pathogen frequently associated with chronic and hard-to-treat infections, particularly in burn units and immunocompromised patients. Its intrinsic and acquired resistance to multiple antibiotics poses a major therapeutic challenge. While ZnO nanoparticles conjugated with thiosemicarbazone (TSC) have shown promise in general antimicrobial applications, their potential for simultaneously inhibiting biofilm formation and pyocyanin production—key virulence factors—in clinical P. aeruginosa strains remains unexplored. In this study, ZnO nanoparticles were synthesized via a hydrothermal route and conjugated with a glutamine-modified TSC ligand (ZnO@Glu-TSC) to enhance their antimicrobial efficacy. The nanoconjugate was comprehensively characterized using UV–Vis spectroscopy, X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDX). Functional evaluations were conducted against clinical isolates of P. aeruginosa, including minimum inhibitory concentration (MIC), fractional inhibitory concentration (FIC) index, biofilm inhibition, and pyocyanin suppression assays. ZnO@Glu-TSC nanoparticles exhibited a sharp UV–Vis absorption peak at 380 nm with a band gap of 3.26 eV, and XRD confirmed a hexagonal wurtzite structure with an average crystallite size of ~ 19.8 nm. The nanoconjugate demonstrated significantly enhanced antibacterial activity with MIC values ranging from 128 to 512 µg/mL and synergistic effects in 70% of clinical isolates (FIC ≤ 0.5, p < 0.01). Biofilm inhibition assays revealed an 80% reduction in biomass (OD values approaching those of the negative control), while pyocyanin production decreased by more than 75% at 512 µg/mL (p < 0.001). These results represent the first demonstration of ZnO@Glu-TSC's dual antivirulence action against clinical P. aeruginosa strains, underscoring its therapeutic promise as a potent, multi-targeted nanoantimicrobial candidate and warranting further development for translational nanomedicine applications in combating persistent infections.