Research of toxic behavior of copper nanoparticles: effect on electrosurface and biochemical parameters of bacterial cells

T. Gruzina, L. Rieznichenko, L. Yakubenko, V. I. Podolska, N. I. Grishchenko, Z. Ulberg, S. Dybkova
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Abstract

This research is aimed at the investigation the electrosurface and biochemical parameters of bacterial cells B. cereus B4368, L. plantarum, E. coli K-A, P. fluorescens B5040 under the influence of copper in ionic form and as nanoparticles in order to determine the nature and level of their toxic effect on bacteria. Copper nanoparticles synthesized in aqueous solution with NaBH4 and stabilized with dextran were used. Changes in membrane transport parameters were assessed by the value of ATPase activity; changes in transmembrane potential were assessed by the method of penetrating tetraphenylphosphonium cations (TPP+); and bacterial integrity was assessed by UV spectroscopy of cellular metabolites. A concentration-dependent inhibition of the membrane ATPase reaction and dissipation of the transmembrane potential under the action of both forms of copper was found, and the inhibitory effect in the case of the nanoparticles was on average 20 % higher than in the ionic form. As a result of heterocoagulation of dextran-stabilized copper nanoparticles and bacteria, a decrease in the negative ξ - potential of bacteria was observed, which was 40 % more effective under the action of copper nanoparticles compared to Cu2+ ions. The most significant changes in membrane parameters were observed in the range 10–60 μM of copper concentrations. With B. cereus B4368 cells taken as an example, we found a violation of the barrier function of their cell membrane under the influence of both copper preparations. In the case of copper nanoparticles, nucleic acid leakage from the bacterial cytoplasm was detected, which was confirmed by the absorption band at 260 nm. The results obtained indicate a high level of sensitivity of the studied electrosurface and biochemical parameters of bacterial cells to the effects of ionic and nanoparticle copper, which allows them to be used as indicators of the toxicity of metal nanoparticles in the development of metal-containing probiotic preparations.
铜纳米颗粒毒性行为的研究:对细菌细胞电表面和生化参数的影响
本研究旨在研究离子铜和纳米铜对蜡样芽孢杆菌B4368、植物乳杆菌、大肠杆菌K-A、荧光杆菌B5040等细菌细胞的电表面和生化参数的影响,以确定它们对细菌的毒性作用的性质和水平。用NaBH4在水溶液中合成铜纳米粒子,用右旋糖酐稳定。通过atp酶活性值评估膜运输参数的变化;采用穿透四苯基磷离子(TPP+)法测定跨膜电位的变化;并用紫外光谱法测定细胞代谢物的细菌完整性。在两种形式的铜的作用下,膜atp酶反应和跨膜电位的耗散都有浓度依赖性的抑制,并且纳米粒子的抑制效果比离子形式的平均高20%。由于右旋糖酐稳定的铜纳米粒子与细菌的异凝聚作用,观察到细菌的负ξ电位下降,与Cu2+离子相比,铜纳米粒子的作用效率提高了40%。铜浓度在10 ~ 60 μM范围内,膜参数变化最为显著。以蜡样芽孢杆菌B4368细胞为例,我们发现两种铜的制备都破坏了其细胞膜的屏障功能。在铜纳米粒子的情况下,检测到核酸从细菌细胞质中泄漏,并通过260 nm的吸收带证实了这一点。结果表明,所研究的细菌细胞的电表面和生化参数对离子铜和纳米铜的影响具有很高的敏感性,这使得它们可以作为金属纳米颗粒毒性的指标,用于含金属益生菌制剂的开发。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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