Abstract
Recently, nanocomposite materials containing nanoparticles of metals such as silver, copper and zinc oxide have attracted most attention due to their pronounced pharmacological properties, such as antimicrobial, antiviral, antiinflammatory, immunomodulatory ability and high stability in extreme conditions. Polyelectrolyte complexes based on polymers of natural origin, namely polysaccharides of chitosan and pectin, which can stabilize nanoparticles of a smaller size than individual polymers have significant potential for creation of silver-containing nanocomposites. The aim of this article is to study the antimicrobial and antiviral activity of silver-containing nanocomposites based on polyelectrolyte complexes. Methods. Peculiarities of the structural organization of silver-containing nanocomposites were investigated by the method of wide-angle scattering on an XRD-7000 diffractometer. The morphology of Ag nanoparticles in polymer matrixes were studied by transmission electron microscopy method (transmission electron microscope JEOL 100 CXII). The antimicrobial activity of silver-containing nanocomposites was determined by agar diffusion assays against opportunistic pathogens Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Candida albicans. Cytotoxicity and antiviral activity were investigated using the MTT method and staining by gentian violet. Results. Analysis of wide-angle X-ray diffractograms of silver-containing nanocomposites based on polyelectrolyte complexes Na-CMC (pectin) – chitosan showed that at reduction of Ag+ ions to metallic silver, there are two low-intensity diffraction maxima at 2θm ~ 380 and 440 in the diffractograms. These maxima correspond to the crystallographic planes of the face-centered cubic lattice of silver, are characterized by indices (111) and (200), respectively, and confirm the presence of metallic silver in the polymer system. Analysis of micrographs of silver-containing nanocomposites based on Na-CMC and chitosan showed that larger nanoparticles are formed with increasing the molecular weight of chitosan. The dependence of the size of silver nanoparticles on the wavelength of ultraviolet radiation at reduction of silver ion in polyelectrolyte-metal complexes Na-CMC–Ag+–chitosan of low molecular weight was also revealed. In particular, smaller particles are formed under irradiation by light with a shorter wavelength (λ =254 nm) than at λ=365 nm. Silver-containing nanocomposites Na-СMC-Ag-chitosan and pectin citrus-Ag-chitosan, obtained by reduction of Ag+ ions under ultraviolet irradiation at a wavelength λ = 365 nm and λ = 254 nm, exhibit high antimicrobial activity against the test cultures of microorganisms S. aureus, E. coli, and P. aeruginosa C. albicans. No significant dependence of antimicrobial activity on the molecular weight of the studied samples was noted: the obtained data were within close limits and had close values. In addition, no dependence of antimicrobial activity on the type of investigated test cultures of microorganisms was observed either. Nanocomposites based on Na-CMC-chitosan (λ = 365 nm) inhibited infection titer HSV-1 by (3.72–5.45) lgTCID50/mL, whereas the decrease in titer during incubation with samples based on citrus pectin-chitosan was within (2.39–2.42) lgTCID50/mL. A dose-dependent relationship between molecular weight of chitosan and reduction of infection titer was observed. It was found that silver-containing nanocomposites formed by reduction of silver ions in polyelectrolyte-metal complexes under ultraviolet radiation of different wavelengths had no cytotoxic effect on cells of MDCK and BHK. Conclusions. The investigated silver-containing nanocomposites based on Na-CMC (pectin)-chitosan polyelectrolyte complexes show antimicrobial activity against test cultures of S. aureus, E. coli P. aeruginosa, and C. albicans along with antiviral activity against herpes simplex virus type 1 and influenza virus. It was established that the obtained nanocomposites did not show a cytotoxic effect on MDCK and BHK cells. The obtained data allow us to assert that investigated silver-containing nanocomposites are promising antimicrobial means for the development of new effective strategies against microorganisms and viruses and improvement of the population health.
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