Silver nanoparticles (Ag-NPs) possess excellent antibacterial properties and are considered to be an alternative material for treating antibiotic-resistant bacteria. The present study was aimed at enhancing the antibacterial efficiency of Ag-NPs using visible laser light against Escherichia coli and Staphylococcus aureus in vitro. Four concentrations of Ag-NPs (12.5, 25, 50, and 100 μg/ml), synthesized by the chemical reduction method, were utilized to conduct the antibacterial activity of prepared Ag-NPs. The antibacterial efficiencies of photoactivated Ag-NPs against both bacteria were determined by survival assay after exposure to laser irradiation. The mechanism of interactions between Ag-NPs and the bacterial cell membranes was then evaluated via scanning electron microscopy (SEM) and reactive oxygen species analysis to study the cytotoxic action of photoactivated Ag-NPs against both bacterial species. Results showed that the laser-activated Ag-NP treatment reduced the surviving population to 14% of the control in the E. coli population, while the survival in the S. aureus population was reduced to 28% of the control upon 10 min exposure time at the concentration of 50 μg/ml. However, S. aureus showed lower sensitivity after photoactivation compared to E. coli. Moreover, the effects depended on the concentration of Ag-NPs and exposure time to laser light. SEM images of treated bacterial cells indicated that substantial morphological changes occurred in cell membranes after treatment. The results suggested that Ag-NPs in the presence of visible light exhibit strong antibacterial activity which could be used to inactivate harmful and pathogenic microorganisms.

Aim: This work reports a new method for the use of lasers for the selective killing of bacteria targeted using light-absorbing Silver nanoparticles (Ag-NPs) conjugated with a specific antibody against the Gram-positive bacterium Staphylococcus aureus (S. aureus). Methods and Results: Ag-NPs were synthesized using a chemical reduction method and characterized with respect to their surface plasmon resonance, surface morphology via transmission electron microscopy (TEM) and dynamic light scattering (DLS). The bacterial surface was targeted using 20 nm Ag-NPs conjugated with an anti-protein A antibody. Labelled bacteria were irradiated with blue visible laser at 2Á04 W/cm 2 . The antibacterial activity of functionalized Ag-NPs was investigated by fluorescence microscopy after irradiation, and morphological changes in S. aureus after laser treatment were assessed using scanning electron microscopy (SEM). The laser-irradiated, functionalized Ag-NPs exhibited significant bactericidal activity, and laserinduced bacterial damage was observed after 10 min of laser irradiation against S. aureus. The fluorescence microscopic analysis results supported that bacterial cell death occurred in the presence of the functionalized Ag-NPs. Conclusions: The results of this study suggest that a novel method for the preparation of functionalized nanoparticles has potential as a potent antibacterial agent for the selective killing of resistant disease-causing bacteria. Significance and Impact of the Study: This study shows that Ag-NPs functionalized with a specific antibody, could be used in combination with laser radiation as a novel treatment to target resistant bacterial and fungal pathogens with minimal impact on normal microflora.