Estimation of cytotoxicity and genotoxicity of selenium nanoparticles stabilized with polyvinylpyrrolidone

E. A. Tsiganovich, S. Dybkova, E. A. Sirik, A. P. Golodyuk, V. A. Prokopenko, A. M. Zhovnir
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Abstract

SeNPs lead to increased production of antibodies and anti-inflammatory cytokines, respiratory activity of lymphocytes and peritoneal macrophages and survival of animals. The goal of the work was to determine the biosafety of promising SeNPs for their using in the biotechnology of veterinary immunological means. Materials and methods. SeNP synthesis performed by reducing selenic acid with ascorbic acid in the presence of polyvinylpyrrolidone. Three sols with SeNP with different selenium concentrations were made: № 1 – 0.05; № 2 – 0.5; № 3 – 0.25 mmol/l. The test object in evaluation toxic effects was the calf kidney cell culture (MDBK). SeNP cytotoxicity was assessed in vitro by cell viability with trypan blue dye. Genotoxicity estimation in vitro has been analyzed by the method of DNA comets in alkaline conditions. Results and discussions. Morphological characteristics of SeNPs determined by electron microscopy. It was found that in the sample № 1 spherical particles have a size of 20-80 nm, № 2 – 70-90 nm and № 3 – 80-90 nm. The results of cytotoxicity studies on test cells showed that at a dilution of 1:50 the survival rate of cells in samples №2 and №3 were 99%. Obtained results of genotoxicity studies showed that SeNP samples in dilution 1:50 were not genotoxic and may have a protective effect. Conclusions and prospects for further research. Studies performed to evaluate biosafety of SeNPs indicate the absence of cytotoxic and genotoxic effects on test cells. Therefore, the synthesized SeNPs are safe and biocompatible and can be used for manufacturing of experimental veterinary immunological means.
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聚乙烯吡咯烷酮稳定纳米硒的细胞毒性和遗传毒性评价
SeNPs导致抗体和抗炎细胞因子的产生增加,淋巴细胞和腹膜巨噬细胞的呼吸活性增加,动物存活率增加。这项工作的目的是确定有前途的SeNPs的生物安全性,用于兽医免疫手段的生物技术。材料和方法。在聚乙烯吡咯烷酮的存在下,用抗坏血酸还原硒酸合成SeNP。制备了三种不同硒浓度的SeNP溶胶:№1 - 0.05;№2 - 0.5;3 - 0.25 mmol/l。评价其毒性作用的试验对象为小牛肾细胞培养物(MDBK)。用台盼蓝染色法测定SeNP体外细胞毒性。在碱性条件下,用DNA彗星法对体外遗传毒性进行了分析。结果和讨论。用电子显微镜测定SeNPs的形态特征。结果发现,样品№1中的球形颗粒尺寸为20-80 nm,№2为70-90 nm,№3为80-90 nm。对试验细胞的细胞毒性研究结果表明,在1:50的稀释下,样品№2和№3的细胞存活率为99%。已获得的遗传毒性研究结果表明,稀释1:50的SeNP样品没有遗传毒性,可能具有保护作用。结论及进一步研究的展望。评估SeNPs生物安全性的研究表明,对测试细胞没有细胞毒性和基因毒性作用。因此,合成的SeNPs具有安全性和生物相容性,可用于制备实验性兽医免疫工具。
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