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Updated: Dec 23, 2025

Evaluation of Antimicrobial Activities of Nanoparticles and Nanostructured Surfaces In Vitro
Published on: April 21, 2023
Efflux pump inhibitory activity of biologically synthesized silver nanoparticles against multidrug-resistant
Reyhaneh Behdad1, Minoo Pargol1, Amir Mirzaie2
1Department of Biology, Varamin-Pishva Branch, Islamic Azad University, Varamin, Iran.
Abstract:
This study was carried out to investigate the possible efflux pump inhibitory activity of biologically synthesized silver nanoparticles (AgNPs) against multidrug-resistant (MDR) Acinetobacter baumannii isolates. In this study, the physicochemical characteristics of synthesized AgNPs were investigated using scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), and Fourier transform infrared spectrophotometer (FTIR) methods. Subsequently, MDR A. baumannii isolates were recovered from clinical samples and the phenotypic cartwheel efflux assay and polymerase chain reaction (PCR) were used to elucidate the possible presence of efflux pump in MDR strains. After treatment of MDR strains with sub-minimum inhibitory concentration (MIC) concentration of AgNPs, the expression level of efflux pump genes was evaluated using a quantitative real-time PCR technique. The synthesized AgNPs had a spherical nanostructure, with mean size 38.89 nm, according to SEM and TEM data. XRD and FTIR results confirmed the synthesis of AgNPs. The results of PCR revealed that among 50 strains, 12 A. baumannii strains had efflux pump genes and the expression level of AdeA, AdeC, AdeS, AdeR, AdeI, AdeJ, and AdeK efflux pump genes was downregulated significantly after the treatment with AgNPs. In addition, the inhibitory effect of AgNPs on efflux pumps can be detected when the MIC of ethidium bromide (EtBr) with AgNPs is lower than that of EtBr alone. According to the results, the biologically synthesized AgNPs exhibit efflux pumps inhibitory activity, which may be one of the possible mechanisms of their antibacterial activity against MDR A. baumannii strains.
Insights
Biologically synthesized silver nanoparticles (AgNPs) show efflux pump inhibitory activity against multidrug-resistant (MDR) Acinetobacter baumannii. This suggests AgNPs may combat MDR bacteria by downregulating efflux pump genes.
Area of Science:
- Nanotechnology
- Microbiology
- Biochemistry
Background:
- Multidrug-resistant (MDR) Acinetobacter baumannii poses a significant clinical challenge.
- Efflux pumps are a major mechanism contributing to MDR in bacteria.
- Novel strategies are needed to overcome antibiotic resistance.
Purpose of the Study:
- To investigate the efflux pump inhibitory activity of biologically synthesized silver nanoparticles (AgNPs).
- To evaluate the effect of AgNPs on efflux pump gene expression in MDR A. baumannii.
Main Methods:
- Physicochemical characterization of AgNPs using SEM, TEM, XRD, and FTIR.
- Identification of efflux pump genes in MDR A. baumannii using PCR.
- Quantitative real-time PCR to assess gene expression after AgNP treatment.
- Ethidium bromide (EtBr) synergy assay to confirm efflux pump inhibition.
Main Results:
- Synthesized AgNPs exhibited spherical nanostructures (mean size 38.89 nm).
- AgNP treatment significantly downregulated the expression of key efflux pump genes (AdeA, AdeC, AdeS, AdeR, AdeI, AdeJ, AdeK).
- AgNPs reduced the minimum inhibitory concentration (MIC) of EtBr, indicating efflux pump inhibition.
Conclusions:
- Biologically synthesized AgNPs possess significant efflux pump inhibitory activity against MDR A. baumannii.
- This inhibitory activity is a potential mechanism underlying the antibacterial effects of AgNPs.
- AgNPs represent a promising therapeutic agent to combat MDR bacterial infections.
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