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Silver nanoparticles are widely used in different fields of medicine despite the lack of information on their influence on animal's reproductive system, mammalian gametes and embryos. We investigated the effect of different concentrations of silver nanoparticles (0, 0.01, 0.1, 1 and 10 µg/mL) on maturation of rabbit's oocytes co-culture with granulosa cells in vitro. For this purpose, we synthesized small (11.28±0.32 nm) spherical silver nanoparticles with different composite agents: polyvinylpyrrolidone and bovine serum albumin. Our results have shown that silver nanoparticles at the concentration of 10 µg/ml inhibited granulosa cells proliferation, but did not influence the oocytes maturation to metaphase-2. The loss of granulosa cells viability was confirmed by the release of calcium and lactate dehydrogenase in the culture medium. Analysis of the data showed that silver nanoparticles in concentration of 0-10 µg/mL did not influence on progesterone and cholesterol concentration in culture medium. We have hypothesized that less toxic effect of silver nanoparticles on oocytes is caused by the presence of zona pellucida with different mechanisms of cellular uptake.

Three-dimensional (3D) cultures models may more accurate representation of the in vivo environment than two-dimensional (2D) cultures while maintaining the cytoarchitecture of in situ tissue that supports cells differentiation or maturation [1, 2]. Cell adhesion is depended upon surface hydrophilicity, surface charge density, surface morpho logy, specific chemical groups present on the surface of the scaffold [3]. Given that surface chemistry is crucial for the biocompatibility of the nanolayers, specific surface modifications are used with different polymers [4, 5]. In particular, there is an increased interest in the polymeric surfaces which can change their affinity towards proteins and cells under external stimuli [6, 7] and therefore have potential applications in biology and medicine. Despite various investigations, specific and complex mechanisms govern the reactions that occur between the biomaterial and the cellular environment are still incomplete understanding. The objective of this study was to establish and comparison cells line B16F10 viability cultured onto different coatings. We used surfaces obtained by grafting APTES, dextran, albumin and their combinations to the surface of the modified glass plates. Materials and Methods Preparation of coatings. Glass plates (2020) were dipped into 0.2 % (w/w) methanolic solution of (3-aminopropyl)triethoxysilane (APTES) for 24 h. After the incubation, loosely-attached silane molecules were removed with methanol in Soxhlet's apparatus. Then the plates functionalized with APTES

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The activity of dissolved enzyme preparations during storage decreases, what leads to the loss of their biological activity and, as a result, reduces the effectiveness of the drugs. Therefore, the development of compositions that are able to maintain high activity of the hormone in dissolved form during long-term storage is relevant. The results of studies have shown that using sucrose as a stabilizing component for maintain gonadotropin activity is effective. It was found that during eight weeks of storage the best results on the preservation of gonadotropin activity during storage at 40°C were obtained in samples containing 75 mg/ml of sucrose compared to the sample of the control group. However, the highest gonadotropin activity was found when — 10 mg/ml L-lysine and 75 mg/ml sucrose were used as stabilizers. Studies of the dynamics of gonadotropin activity during long-term storage at 18–20°C showed that the addition of L-lysine and sucrose as stabilizing substances in the form of liposomal emulsion increases the preservation of chorionic hormone activity for 2 weeks of storage by 11.4% compared to similar composition pharmacological composition of the drug in aqueous form.

In reproductive medicine it is important to determine the quality of embryo development without interference in their function and viability. The surface plasmon resonance of silver nanoparticles makes them promising candidates for optical sensing, molecular labeling and imaging applications. Furthermore unique optical properties of silver nanoparticles provide an opportunity to use them as real time analytic tools in living state especially for observation of dynamic processes in gametes and embryos. The main aim of the study was to investigate the physicochemical properties and biological activities of novel silver nanoparticles with prospect of their use for the determining the quality of embryo development. For this purpose, we investigated the optical properties of new silver nanoparticles in biological mediums during cultivation and their influence on rabbit's embryos development in vitro. The physicochemical and biological properties of novel silver nanoparticles were compared with silver nanoparticles identical in size and shapes but with different chemical surfaces modifications by polyvinylpyrrolidone and bovine serum albumin. The results suggest that silver nanoparticles with hyaluronic acid were disintegrated with the formation of new complexes with proteins in biological mediums. This property with strong optical surface plasmon resonance of novel silver nanoparticles with hyaluronan makes them promising candidates in diagnostic area and gives reasons to explore them as biomarkers of target molecules. Nevertheless novel silver nanoparticles with hyaluronan at the concentrations of 0.1-1 µg/ml have no toxic effect on rabbit's embryos development and can be successfully applied in reproductive medicine.

The more stable among the tested samples were samples with saccharose in the concentration of 50–75 mg per cm3. While adding of L- lysine to samples the most stable activity was discovered in the experimental series of samples with the content of lysine of 10 mg per cm3 – activity increased by 54 % as compared to theoretical initial activity of HCG during 8 weeks. While storing gonadotropin with L-glycine fluctuations of hormone activity in all series of samples were observed. Adding of 0.2 mg per cm3 of L-glycine had a more expressed stabilizing effect. Adding of 0.2 mg per cm3 of L-methionine produced relatively high and stable activity of gonadotropin during the 6 weeks storage. Adding of 0.25 mg / cm3 of L- glycine and 75.50 mg / cm3 of saccharose to experimental samples during 2 weeks at 40 °C provided 69.8 % and 60.7 % saving activity of hCG respectively. Activity of gonadotropin in a series of samples with the addition of L- glycine and mannitol was significantly lower and at the end of the study was at an appropriate rate with the control series models. The highest activity of gonadotropin was detected while adding fillers – 10 mg / cm3 L-lysine and 75 mg / cm3 saccharose and mannitol – to recipes as a stabilizer.