Evaluation of biosynthesized silver nanoparticles effects on expression levels of virulence and biofilm‐related genes of multidrug‐resistant Klebsiella pneumoniae isolates

Abstract

AbstractThe emergence of multidrug‐resistant (MDR) strains of Klebsiella pneumoniae is associated with high morbidity and mortality due to limited treatment options. This study attempts to biologically synthesize silver nanoparticles (AgNPs) and investigate their effect on expression levels of virulence and biofilm‐related genes in clinically isolated K. pneumoniae. In this study, biofilm formation ability, antibiotic resistance pattern, extended‐spectrum β‐lactamases (ESBLs), and carbapenemases production were investigated for 200 clinical isolates of K. pneumoniae using phenotypic methods. Polymerase chain reaction (PCR) was used to detect virulence and biofilm‐related genes, ESBL‐encoding genes, and carbapenem resistance genes. AgNPs were synthesized using the bio‐reduction method. The antibacterial effects of AgNPs were investigated by microdilution broth. In addition, the cytotoxic effect of AgNPs on L929 fibroblast cell lines was determined. The effects of AgNPs on K. pneumoniae virulence and biofilm‐related genes (fimH, rmpA, and mrkA) were determined using quantitative real‐time PCR. Thirty percent of the isolates produced a strong biofilm. The highest and lowest levels of resistance were observed against amoxicillin/clavulanic acid (95.4%) and tigecycline (96%), respectively. About 31% of isolates were considered positive for carbapenemases, and 75% of the isolates produced an ESBLs enzyme. Different frequencies of mentioned genes were observed. The synthesized AgNPs had a spherical morphology and varied in size. AgNPs inhibited the growth of MDR K. pneumoniae at 128 µg/ml. In addition, AgNPs downregulated the expression of fimH, rmpA, and mrkA genes by 10, 7, and 14‐fold, respectively (p < 0.05), also exerted no cytotoxic effect on L929 fibroblast cell lines. It was revealed that AgNPs lead to a decrease in expression levels of virulence and biofilm‐related genes; therefore, it was concluded that AgNPs had an excellent antibacterial effect on MDR K. pneumoniae.