Effect of Metal Nanoparticles on EBV-Associated Cell Culture

Q4 Biochemistry, Genetics and Molecular Biology Mikrobiolohichnyi zhurnal Pub Date : 2023-02-17 DOI:10.15407/microbiolj84.05.030
S. Zahorodnia, K. Naumenko, O. Zaychenko, P. Zaremba, G. Baranova, A. V. Holovan'
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Abstract

Today, one of the topical areas of research is the search for antiviral drugs to fi ght against virus-associated oncological manifestations. One of the viruses for which a role in the transformation of cells is proved is Epstein-Barr virus (EBV), which is associated with a variety of lymphoproliferative diseases. The use of drugs that not only inhibit the replication of the virus but also stimulate the elimination of tumor cells is important for the treatment of tumors associated with the viruses. The purpose of this work was to investigate the ability of silver and gold nanoparticles to inhibit EBV replication under conditions of chronic infection. Methods. The objects of the study were 5 to 20 nm gold and silver nanoparticles stabilized with tryptophan, sodium dodecyl sulfate, and citrate. Th e investigations were performed in P3HR-1 (virus-productive) lymphoblastoid cells. MTT-assay, neutral red and trypan blue dyeing were used to study cell viability. Antiviral activity was estimated by the real-time polymerase chain reaction (RT-PCR). Transmissive electron microscopy was used to visualize nanoparticle-virus binding. Results. It was found that nanoparticles of silver and gold stabilized by tryptophan and citrate were low-toxic for the used cell cultures; the vitality of the cells was in the range of 65—100%. Silver nanoparticles in a citrate buffer were more effective against EBV because the used concentrations inhibited replication of the virus up to 70%. Gold nanoparticles reduced the amount of EBV DNA by a maximum of 16% at the lowest concentration of 0.00001 μg/mL, indicating a dose-dependent effect. The virucidal effect of gold nanoparticles against EBV was shown using transmissive electron microscopy. It was found that the interaction of the virus with 5 nm gold nanoparticles for 2 hr leads to damage of EBV virion, which indicates their virus-static effect. Conclusions. Thus, the cytotoxic and antiviral activity of silver and gold nanoparticles in different stabilizers was analyzed. Citrate buffer-stabilized silver and gold NPs were more effective against EBV.
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金属纳米颗粒对ebv相关细胞培养的影响
今天,一个热门的研究领域是寻找抗病毒药物来对抗病毒相关的肿瘤表现。在细胞转化中起作用的病毒之一是eb病毒(EBV),它与多种淋巴增生性疾病有关。使用既能抑制病毒复制又能刺激肿瘤细胞消除的药物对于治疗与病毒相关的肿瘤很重要。这项工作的目的是研究银和金纳米颗粒在慢性感染条件下抑制EBV复制的能力。方法。研究对象为5 ~ 20 nm的金、银纳米粒子,用色氨酸、十二烷基硫酸钠和柠檬酸盐稳定。研究在P3HR-1(产生病毒)淋巴母细胞样细胞中进行。采用mtt法、中性红和台盼蓝染色研究细胞活力。采用实时聚合酶链反应(RT-PCR)测定抗病毒活性。利用透射电子显微镜观察纳米颗粒与病毒的结合。结果。经色氨酸和柠檬酸稳定的银和金纳米颗粒对使用过的细胞培养物具有低毒作用;细胞活力在65-100%之间。柠檬酸缓冲液中的银纳米颗粒对EBV更有效,因为所使用的浓度可抑制病毒复制高达70%。在最低浓度为0.00001 μg/mL时,金纳米颗粒最多可使EBV DNA减少16%,且呈剂量依赖性。透射电镜观察了金纳米颗粒对eb病毒的毒力。结果表明,5 nm的金纳米粒子与EBV病毒粒子相互作用2小时,可导致EBV病毒粒子的损伤,表明其具有病毒静态作用。结论。因此,我们分析了银和金纳米颗粒在不同稳定剂中的细胞毒性和抗病毒活性。柠檬酸缓冲稳定的银和金NPs对EBV更有效。
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Mikrobiolohichnyi zhurnal
Mikrobiolohichnyi zhurnal Medicine-Microbiology (medical)
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