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.
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.
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.
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