Biofilm inhibition of multidrug-resistant Pseudomonas aeruginosa using green-synthesized silver nanoparticles and colistin

We aimed to investigate the synergistic effects of colistin and green-synthesized silver nanoparticles on the biofilm formation and expression of Quorum Sensing regulated and related genes in clinical isolates of P. aeruginosa. Ten clinical P. aeruginosa isolates collected from patients with burn wound infections were investigated. The antibiotic sensitivity pattern of the isolates was determined using disk diffusion and microbroth dilution tests. The silver nanoparticles (AgNPs) were synthesized using propolis and characterized. The microtiter plate method and scanning electron microscopy (SEM) were used to evaluate the synergistic effects of colistin and silver nanoparticles combination (AgNPs@CL) on the inhibition of biofilm formation. The effect of AgNPs@CL on the expression of genes controlled by QS was evaluated using RT-PCR. All isolates were strong biofilm formers. Confronting AgNPs@CL, all isolates were either synergistic or additive and effectively decrease the minimum inhibitory concentration (MIC) and minimum biofilm inhibitory concentration (MBIC) values of Carbapenem-Resistant P. aeruginosa (CRPA) isolates. The SEM analysis corroborated the enhanced biofilm inhibition observed with the combined treatment compared to individual AgNPs or colistin treatments. When exposed to AgNPs@CL, the expression levels of lasI, lasR, rhlI, rhlR, pelA, and pslA genes significantly decreased in P. aeruginosa ATCC 27,853 and clinical isolate No. #354, which displayed synergistic activity. In contrast, with additive activity, clinical isolate No. #30 showed no significant decrease. Targeting critical components of QS could effectively inhibit biofilm production. The results of our study suggest AgNPs@CL as an auxiliary to antibiotic therapy.