Esnaashari Fatemeh, Nikzad Ghazaleh, Zahmatkesh Hossein, Zamani Hojjatolah
Department of Biology, Faculty of Basic Sciences, Lahijan Branch, Islamic Azad University, Lahijan, Guilan, Iran.
Department of Biology, University of Guilan, Rasht, Iran.
Bioorg Chem. 2025 Jul 1;161:108527. doi: 10.1016/j.bioorg.2025.108527. Epub 2025 Apr 26.
The dense biofilm architecture and efflux pump activity play critical roles in Pseudomonas aeruginosa infections by hindering the accumulation and long-term efficacy of antibacterial agents within bacterial cells. The development of engineered nanoparticles capable of penetrating the polysaccharide matrix of biofilms represents a promising strategy for addressing bacterial infections. This is the first report on the synthesis of quercetin-functionalized PEGylated ZnO nanoparticles (ZnO-PEG-QUE NPs) and the evaluation of their anti-biofilm activity against pathogenic strains of P. aeruginosa. The synthesized NPs exhibited spherical shapes with an average size of 59.52 nm. ZnO-PEG-QUE NPs demonstrated biofilm inhibitory levels between 49 % and 67 %, and significantly reduced the production of total exopolysaccharides, alginate, and pellicle by 64.61 %-71.69 %, 30.47 %-45.36 %, and 24.22 %-85.97 %, respectively. ZnO-PEG-QUE NPs not only inhibited early-stage biofilm formation but also disrupted mature biofilms, indicating a dual mode of action against both biofilm development and persistence. Based on our findings, ZnO-PEG-QUE NPs effectively eradicated mature biofilms by 67.2 %-72 % and significantly reduced the metabolic activity and viable cells of preformed biofilms to 34.12 %-55.57 % and 6.25-8.15 log CFU, respectively. Electron and fluorescence microscopy analyses also confirmed the antibiofilm potential of ZnO-PEG-QUE NPs. Furthermore, bacterial adhesion and invasion to HDF cells were significantly diminished in the NP-treated groups. The attenuation of efflux pump activity in the NP-treated strains was confirmed using the EtBr-agar cartwheel assay. Taken together, these findings highlight the therapeutic potential of ZnO-PEG-QUE NPs as a novel and effective strategy to combat biofilm-associated infections, warranting further investigation in preclinical models.
致密的生物膜结构和外排泵活性在铜绿假单胞菌感染中起着关键作用，它们会阻碍抗菌剂在细菌细胞内的积累以及长期疗效。开发能够穿透生物膜多糖基质的工程纳米颗粒是应对细菌感染的一种有前景的策略。这是关于槲皮素功能化聚乙二醇化氧化锌纳米颗粒（ZnO-PEG-QUE NPs）的合成及其对铜绿假单胞菌致病菌株的抗生物膜活性评估的首次报告。合成的纳米颗粒呈球形，平均尺寸为59.52纳米。ZnO-PEG-QUE NPs表现出49%至67%的生物膜抑制水平，并分别显著降低了总胞外多糖、藻酸盐和菌膜的产生，降幅分别为64.61% - 71.69%、30.47% - 45.36%和24.22% - 85.97%。ZnO-PEG-QUE NPs不仅抑制早期生物膜形成，还能破坏成熟生物膜，表明其对生物膜形成和持续存在具有双重作用模式。基于我们的研究结果，ZnO-PEG-QUE NPs能有效根除67.2% - 72%的成熟生物膜，并将预先形成的生物膜的代谢活性和活细胞分别显著降低至34.12% - 55.57%和6.25 - 8.15 log CFU。电子显微镜和荧光显微镜分析也证实了ZnO-PEG-QUE NPs的抗生物膜潜力。此外，纳米颗粒处理组中细菌对人皮肤成纤维细胞（HDF）的黏附和侵袭显著减少。使用溴化乙锭 - 琼脂车轮试验证实了纳米颗粒处理菌株中外排泵活性的减弱。综上所述，这些发现突出了ZnO-PEG-QUE NPs作为对抗生物膜相关感染的一种新颖且有效策略的治疗潜力，值得在临床前模型中进一步研究。
