Effect of graphene oxide nanoparticles on apoptosis of T-lymphocytes and Jurkat cells

D. Usanina, S. Uzhviyuk, S. Zamorina
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Abstract

Graphene and its derivatives are materials with unique physicochemical properties. A detailed study of these materials allows to consider them prospective biomedical agents for targeted drug and gene delivery, photothermal therapy of cancer, bioimaging, etc. However, this requires a comprehensive studies of their effects on the body tissues, including cells of the immune system. The aim of our research was to stydy the effects of nanoparticles based on pegylated graphene oxide (GO) upon apoptosis of T lymphocytes derived from blood of healthy donors and Jurkat 5332 cell line. Comparison of these cells will extend our knowledge of the effects of nanomaterials on the cells, and to respond the question, what results obtained with continuous cell lines are valid for normal non-malignant cells. In this work, we used GO nanoparticles (100-200 nm, 1-5 m) coated with linear (LP-GO) and branched (BP-GO) polyethylene glycol (PEG). The cells were cultured for 24 hours at 37 C and 5% CO2with nanoparticles at concentrations of 5 and 25 g/mL. Viability and early and late apoptosis of incubated Jurkat cells and CD3+cells from healthy donors were assessed by flow cytometry. It was found that the small nanoparticles coated with linear PEG at high concentrations (25 g/mL) could significantly reduce the number of live cells and increase the number of cells in late apoptosis. At the same time, large nanoparticles coated with branched PEG at high concentrations (25 g/mL) increased the percentage of T cells in early apoptosis. Meanwhile, the GO nanoparticles at both concentrations did not affect the viability and apoptosis of Jurkat cells, regardless of the size, concentration, and type of surface function of the particles. The obtained results suggest that GO nanoparticles exert different effects upon normal and malignant lymphocytes of T lineage. One may assume that these discrepancies could be explained by greater resistance of tumor cells compared to normal T cells. These findings suggests that studies of nanomaterials upon living cells should not be limited to experiments on cell lines, since their properties may significantly differ from those of non-malignant cells.
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氧化石墨烯纳米颗粒对t淋巴细胞和Jurkat细胞凋亡的影响
石墨烯及其衍生物是具有独特物理化学性质的材料。对这些材料的详细研究可以考虑将其用于靶向药物和基因递送、癌症光热治疗、生物成像等生物医学制剂。然而,这需要全面研究它们对身体组织的影响,包括免疫系统的细胞。我们的研究目的是研究基于聚乙二醇化氧化石墨烯(GO)的纳米颗粒对来自健康供者血液和Jurkat 5332细胞系的T淋巴细胞凋亡的影响。这些细胞的比较将扩展我们对纳米材料对细胞的影响的认识,并回答这样一个问题,用连续细胞系获得的结果对正常的非恶性细胞有效。在这项工作中,我们使用了涂有线性(LP-GO)和支链(BP-GO)聚乙二醇(PEG)的氧化石墨烯纳米颗粒(100-200 nm, 1-5 m)。细胞在37℃和5% co2条件下培养24小时,纳米颗粒浓度分别为5和25 g/mL。流式细胞术检测健康供体Jurkat细胞和CD3+细胞的活力及早期和晚期凋亡情况。结果发现,经线状聚乙二醇包被的小纳米颗粒在高浓度(25 g/mL)下可显著减少活细胞数量,增加晚期凋亡细胞数量。同时,高浓度(25 g/mL)包被支链聚乙二醇的大纳米颗粒增加了T细胞早期凋亡的百分比。同时,无论氧化石墨烯纳米颗粒的大小、浓度和表面功能类型如何,两种浓度的氧化石墨烯纳米颗粒都不会影响Jurkat细胞的活力和凋亡。结果表明氧化石墨烯纳米颗粒对T系正常淋巴细胞和恶性淋巴细胞的作用不同。人们可能认为这些差异可以解释为肿瘤细胞比正常T细胞具有更大的抵抗力。这些发现表明,纳米材料在活细胞上的研究不应局限于细胞系上的实验,因为它们的特性可能与非恶性细胞的特性有很大不同。
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