In Vitro Hemocompatibility and Toxic Mechanism of Graphene Oxide on Human Peripheral Blood T Lymphocytes and Serum Albumin
In Vitro Hemocompatibility and Toxic Mechanism of Graphene Oxide on Human Peripheral Blood T Lymphocytes and Serum Albumin
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DOI：
10.1021/am505084s
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发表时间：
2014-11-26
影响因子：
9.5
通讯作者：
Chen, Yanyan
中科院分区：
文献类型：
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作者：
Ding, Zhijia;Zhang, Zhijun;Chen, Yanyan
氧化石墨烯（GO）作为一种生物医用材料显示出巨大的应用潜力。然而，它与血液成分的相互作用尚未得到很好的理解。在这项工作中，我们评估了原始GO（p-GO）和功能化GO（GO-COOH和GO-PEI）对原代人外周血T淋巴细胞和人血清白蛋白（HSA）的毒性，并研究了潜在的毒性机制。我们的结果表明，p-GO和GO-COOH在浓度低于25 μ g mL（-1）时对T淋巴细胞具有良好的生物相容性，但在浓度高于50 μ g mL（-1）时具有显著的细胞毒性。相比之下，GO-PEI甚至在1.6 μ g/mL（-1）时也表现出显著的毒性。进一步的研究表明，虽然p-GO不会进入细胞或损伤细胞膜，但它的存在会导致活性氧（ROS）的增加，中度DNA损伤和T淋巴细胞凋亡。有趣的是，尽管p-GO引起细胞凋亡，但在该过程中观察到对T淋巴细胞免疫应答抑制的影响很小。其毒性机制是p-GO直接与蛋白受体相互作用，抑制其配体结合能力，通过B细胞淋巴瘤-2（Bcl-2）途径导致ROS依赖性被动凋亡。与p-GO相比，GO-COOH对T淋巴细胞的毒性作用相似，但保持正常的ROS水平。一种被提出的毒性机制是GO-COOH抑制蛋白质受体-配体结合，并通过ROS非依赖性机制将被动凋亡信号传递到细胞核DNA。另一方面，GO-PEI通过诱导膜损伤对T淋巴细胞显示严重的血液毒性。对于血浆蛋白HSA，GO-COOH的结合导致最小的构象变化，并且HSA与胆红素的结合能力保持不受影响，而p-GO和GO-PEI的结合对HSA表现出强毒性。这些关于二维纳米材料与生物系统相互作用的研究结果，沿着其作用机制的探讨，将为GO生物医学应用的进一步安全性评价提供必要的支持。
Graphene oxide (GO) has shown tremendous application potential as a biomedical material. However, its interactions with blood components are not yet well understood. In this work, we assess the toxicity of pristine GO (p-GO) and functionalized GO (GO-COOH and GO-PEI) to primary human peripheral blood T lymphocytes and human serum albumin (HSA), and also study the underlying toxic mechanism. Our results indicate that p-GO and GO-COOH have good biocompatibility to T lymphocytes at the concentration below 25 mu g mL(-1), but notable cytotoxicity above 50 mu g mL(-1). By contrast, GO-PEI exhibits significant toxicity even at 1.6 mu g mL(-1). Further investigations show that although p-GO does not enter into the cell or damage the membrane, its presence leads to the increase in reactive oxygen species (ROS), moderate DNA damage, and T lymphocyte apoptosis. Interestingly, little effect on T lymphocyte immune response suppression is observed in this process despite p-GO inflicting cell apoptosis. The toxic mechanism is that p-GO interacts directly with the protein receptors to inhibit their ligand-binding ability, leading to ROS-dependent passive apoptosis through the B-cell lymphoma-2 (Bcl-2) pathway. Compared with p-GO, GO-COOH exhibits a similar toxic effect on T lymphocytes except keeping a normal ROS level. A proposed toxic mechanism is that GO-COOH inhibits protein receptor-ligand binding, and passes the passive apoptosis signal to nucleus DNA through a ROS-independent mechanism. On the other hand, GO-PEI shows severe hematotoxicity to T lymphocytes by inducing membrane damage. For plasma protein HSA, the binding of GO-COOH results in minimal conformational change and HSA's binding capacity to bilirubin remains unaffected, while the binding of p-GO and GO-PEI exhibits strong toxicity on HSA. These findings on the interactions of two-dimensional nanomaterials and biological systems, along with the enquiry of the mechanisms, would provide essential support for further safety evaluation of the biomedical applications of GO.