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      International Journal of Nanomedicine (submit here)

      This international, peer-reviewed Open Access journal by Dove Medical Press focuses on the application of nanotechnology in diagnostics, therapeutics, and drug delivery systems throughout the biomedical field. Sign up for email alerts here.

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      Negatively charged silver nanoparticles with potent antibacterial activity and reduced toxicity for pharmaceutical preparations

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          Abstract

          Background

          The discovery of new solutions with antibacterial activity as efficient and safe alternatives to common preservatives (such as parabens) and to combat emerging infections and drug-resistant bacterial pathogens is highly expected in cosmetics and pharmaceutics. Colloidal silver nanoparticles (NPs) are attracting interest as novel effective antimicrobial agents for the prevention of several infectious diseases.

          Methods

          Water-soluble, negatively charged silver nanoparticles (AgNPs) were synthesized by reduction with citric and tannic acid and characterized by transmission electron microscopy, dynamic light scattering, zeta potential, differential centrifuge sedimentation, and ultraviolet–visible spectroscopy. AgNPs were tested with model Gram-negative and Gram-positive bacteria in comparison to two different kinds of commercially available AgNPs.

          Results

          In this work, AgNPs with higher antibacterial activity compared to the commercially available colloidal silver solutions were prepared and investigated. Bacteria were plated and the antibacterial activity was tested at the same concentration of silver ions in all samples. The AgNPs did not show any significant reduction in the antibacterial activity for an acceptable time period. In addition, AgNPs were transferred to organic phase and retained their antibacterial efficacy in both aqueous and nonaqueous media and exhibited no toxicity in eukaryotic cells.

          Conclusion

          We developed AgNPs with a 20 nm diameter and negative zeta potential with powerful antibacterial activity and low toxicity compared to currently available colloidal silver, suitable for cosmetic preservatives and pharmaceutical preparations administrable to humans and/or animals as needed.

          Most cited references24

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          Antibacterial activity and mechanism of action of the silver ion in Staphylococcus aureus and Escherichia coli.

          The antibacterial effect and mechanism of action of a silver ion solution that was electrically generated were investigated for Staphylococcus aureus and Escherichia coli by analyzing the growth, morphology, and ultrastructure of the bacterial cells following treatment with the silver ion solution. Bacteria were exposed to the silver ion solution for various lengths of time, and the antibacterial effect of the solution was tested using the conventional plate count method and flow cytometric (FC) analysis. Reductions of more than 5 log(10) CFU/ml of both S. aureus and E. coli bacteria were confirmed after 90 min of treatment with the silver ion solution. Significant reduction of S. aureus and E. coli cells was also observed by FC analysis; however, the reduction rate determined by FC analysis was less than that determined by the conventional plate count method. These differences may be attributed to the presence of bacteria in an active but nonculturable (ABNC) state after treatment with the silver ion solution. Transmission electron microscopy showed considerable changes in the bacterial cell membranes upon silver ion treatment, which might be the cause or consequence of cell death. In conclusion, the results of the present study suggest that silver ions may cause S. aureus and E. coli bacteria to reach an ABNC state and eventually die.
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            Silver nanoparticles: partial oxidation and antibacterial activities.

            The physical and chemical properties of silver nanoparticles that are responsible for their antimicrobial activities have been studied with spherical silver nanoparticles (average diameter approximately 9 nm) synthesized by the borohydride reduction of Ag+ ions, in relation to their sensitivity to oxidation, activities towards silver-resistant bacteria, size-dependent activities, and dispersal in electrolytic solutions. Partially (surface) oxidized silver nanoparticles have antibacterial activities, but zero-valent nanoparticles do not. The levels of chemisorbed Ag+ that form on the particle's surface, as revealed by changes in the surface plasmon resonance absorption during oxidation and reduction, correlate well with the observed antibacterial activities. Silver nanoparticles, like Ag+ in the form of AgNO3 solution, are tolerated by the bacteria strains resistant to Ag+. The antibacterial activities of silver nanoparticles are related to their size, with the smaller particles having higher activities on the basis of equivalent silver mass content. The silver nanoparticles aggregate in media with a high electrolyte content, resulting in a loss of antibacterial activities. However, complexation with albumin can stabilize the silver nanoparticles against aggregation, leading to a retention of the antibacterial activities. Taken together, the results show that the antibacterial activities of silver nanoparticles are dependent on chemisorbed Ag+, which is readily formed owing to extreme sensitivity to oxygen. The antibacterial activities of silver nanoparticles are dependent on optimally displayed oxidized surfaces, which are present in well-dispersed suspensions.
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              The inhibitory effects of silver nanoparticles, silver ions, and silver chloride colloids on microbial growth.

              Emerging nanomaterials are of great concern to wastewater treatment utilities and the environment. The inhibitory effects of silver nanoparticles (Ag NPs) and other important Ag species on microbial growth were evaluated using extant respirometry and an automatic microtiter fluorescence assay. Using autotrophic nitrifying organisms from a well-controlled continuously operated bioreactor, Ag NPs (average size=14+/-6 nm), Ag(+) ions (AgNO(3)), and AgCl colloids (average size=0.25 microm), all at 1mg/L Ag, inhibited respiration by 86+/-3%, 42+/-7%, and 46+/-4%, respectively. Based on a prolonged microtiter assay, at about 0.5mg/L Ag, the inhibitions on the growth of Escherichia coli PHL628-gfp by Ag NPs, Ag(+) ions, and AgCl colloids were 55+/-8%, 100%, and 66+/-6%, respectively. Cell membrane integrity was not compromised under the treatment of test Ag species by using a LIVE/DEAD Baclight bacterial viability assay. However, electron micrographs demonstrated that Ag NPs attached to the microbial cells, probably causing cell wall pitting. The results suggest that nitrifying bacteria are especially susceptible to inhibition by Ag NPs, and the accumulation of Ag NPs could have detrimental effects on the microorganisms in wastewater treatment.
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                Author and article information

                Journal
                Int J Nanomedicine
                Int J Nanomedicine
                International Journal of Nanomedicine
                International Journal of Nanomedicine
                Dove Medical Press
                1176-9114
                1178-2013
                2017
                31 March 2017
                : 12
                : 2517-2530
                Affiliations
                [1 ]Nanobiolab, Department of Biotechnology and Bioscience, University of Milano-Bicocca
                [2 ]Biological and Clinical Science Department, University of Milan, Milano
                [3 ]Surgery Department, Breast Unit, IRCCS S Maugeri Foundation, Pavia, Italy
                Author notes
                Correspondence: Davide Prosperi; Miriam Colombo, Department of Biotechnology and Bioscience, University of Milano-Bicocca, Piazza della Scienza 2, 20126 Milano, Italy, Tel +39 02 6448 3388, Fax +39 02 6448 3565, Email davide.prosperi@ 123456unimib.it ; miriam.colombo@ 123456unimib.it
                Article
                ijn-12-2517
                10.2147/IJN.S127799
                5383075
                28408822
                9f20a9c3-9527-4ebf-8f86-649ff06cf1e6
                © 2017 Salvioni et al. This work is published and licensed by Dove Medical Press Limited

                The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License ( http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed.

                History
                Categories
                Original Research

                Molecular medicine
                silver nanoparticles,antibacterial activity,long-term effect,nanoparticle toxicity,phase transfer

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