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S. M. Dybkova, V. I. Podolska, N. I. Gryshchenko, Z. R. Ulberg
«Nanobiocomposite Based on Ultradispersed Silver for the Production of Probiotics»
189–204 (2020)

PACS numbers: 81.07.Pr, 81.16.Fg, 87.15.N-, 87.16.Gj, 87.19.xb, 87.64.-t, 87.85.jj

This report presents the experimental results for nanobiocomposite material (NBC) based on Lactobacillus plantarum cells and ultrafine silver particles, which is fabricated by means of the ‘green synthesis’ method. Synthesized NBC samples are investigated with energy dispersive x-ray spectroscopy analysis, transmission and scanning electron microscopies, and spectroscopy methods. The formation of ultrafine silver particles in a lactobacilli cell wall with an average particles’ size equal to 4–6 nm is confirmed and corresponds to the plasmon absorption band at 390 nm. The strong signal on EDS spectrum at 3 keV from atoms of exogenous silver is observed. That is typical for silver nanoparticle absorption. NBC material is investigated on cytotoxicity and genotoxicity in the range of concentrations (2.5–40.0)\(\cdot\)10\(^{-5}\) µg/ml respectively. Cytotoxicity tests are carried out by the visual growth-rate indicators of monolayer of the eukaryotic cells and by the indicator of viability of the passed eukaryotic cells under microscopic analysis of cells’ monolayer and their staining with 0.4% vital dye of trypan blue. Genotoxicity is evaluated by the method of Comet assay in alkaline conditions in vitro. The culture of calf kidney MDBK is served as the test object when evaluating the cito- and genotoxicity. The full set of experimental investigations of NBC shows the low level of their cytotoxic and genotoxic effects. The lyophilized preparations of probiotic L. plantarum culture with addition of NBC material based on the same culture are made by the freeze-drying method. As estimated, the growth of lyophilized preparations after their rehydration in physiological solution depends on the concentration of added ultrafine silver particles in protective medium. At least, triple increase of L. plantarum colonies is observed for the examples containing 8.0\(\cdot\)10\(^{-5}\) µg/mL of silver nanoparticles (as silver concentration). The effect of significant increase of the viability of the lyophilized L. plantarum cells with the addition of NBC additives based on the same culture, which contain ultrafine silver, can be used for manufacturing of lactobacilli probiotics. Probably, the observed behaviour of NBC fabrication under investigation is due to its prebiotic properties. Besides biogenic silver, this composite containing fragments of bacterial cells, which is served as a source of nutrition for rehydrated probiotic bacteria.

Keywords: nanobiocomposite, silver nanoparticles, probiotics, biosafety, lyophilisation

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