A systematic and detailed study for size-specific antibacterial efficacy of silver nanoparticles (AgNPs) synthesized using a co-reduction approach is presented here. Nucleation and growth kinetics during the synthesis process was precisely controlled and AgNPs of average size 5, 7, 10, 15, 20, 30, 50, 63, 85, and 100 nm were synthesized with good yield and monodispersity. We found the bacteriostatic/bactericidal effect of AgNPs to be size and dose-dependent as determined by the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of silver nanoparticles against four bacterial strains. Out of the tested strains, Escherichia coli MTCC 443 and Staphylococcus aureus NCIM 5201 were found to be the most and least sensitive strains regardless of AgNP size. For AgNPs with less than 10 nm size, the antibacterial efficacy was significantly enhanced as revealed through delayed bacterial growth kinetics, corresponding MIC/MBC values and disk diffusion tests. AgNPs of the smallest size, i.e., 5 nm demonstrated the best results and mediated the fastest bactericidal activity against all the tested strains compared to AgNPs having 7 nm and 10 nm sizes at similar bacterial concentrations. TEM analysis of AgNP treated bacterial cells showed the presence of AgNPs on the cell membrane, and AgNPs internalized within the cells.
Antimicrobial materials with immobilized/entrapped silver nanoparticles (AgNPs) are of considerable interest. There is significant debate on the mode of bactericidal action of AgNPs, and both contact killing and/or ion mediated killing have been proposed. In this study, AgNPs were immobilized on an amine-functionalized silica surface and their bactericidal activity was studied concurrently with the silver release profile over time. This was compared with similar studies performed using colloidal AgNPs and AgCl surfaces that released Ag ions. We conclude that contact killing is the predominant bactericidal mechanism and surface immobilized nanoparticles show greater efficacy than colloidal AgNPs, as well as a higher concentration of silver ions in solution. In addition, the AgNP immobilized substrate was used multiple times with good efficacy, indicating this immobilization protocol is effective for retaining AgNPs while maintaining their disinfection potential. The antibacterial surface was found to be extremely stable in aqueous medium and no significant leaching (∼1.15% of total silver deposited) of the AgNPs was observed. Thus, immobilization of AgNPs on a surface may promote reuse, reduce environmental risks associated with leaching of AgNPs and enhance cost effectiveness.
Hydrogels are water-insoluble crosslinked hydrophilic networks capable of retaining a large amount of water. The present work aimed to develop a novel chitosan-PVA-based hydrogel which could behave both as a nanoreactor and an immobilizing matrix for silver nanoparticles (AgNPs) with promising antibacterial applications. The hydrogel containing AgNPs were prepared by repeated freeze-thaw treatment using varying amounts of the crosslinker, followed by in situ reduction with sodium borohydride as a reducing agent. Characterization studies established that the hydrogel provides a controlled and uniform distribution of nanoparticles within the polymeric network without addition of any further stabilizer. The average particle size was found to be 13 nm with size distribution from 8 to 21 nm as per HR-TEM studies. Swelling studies confirmed that higher amount of crosslinker and silver incorporation inside the gel matrices significantly enhanced the porosity and chain entanglement of the polymeric species of the hydrogel, respectively. The AgNP-hydrogel exhibited good antibacterial activity and was found to cause significant reduction in microbial growth (Escherichia coli) in 12 h while such activity was not observed for the hydrogel without AgNPs.
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