Amrita Centre for Nanosciences and Molecular Medicine, Amrita Institute of Medical, Sciences and Research Centre, Amrita Vishwa Vidyapeetham University, Cochin 682 041, Kerala, India.
Small. 2012 Apr 23;8(8):1251-63. doi: 10.1002/smll.201102393. Epub 2012 Feb 15.
Graphene and its derivatives are being proposed for several important biomedical applications including drug delivery, gene delivery, contrast imaging, and anticancer therapy. Most of these applications demand intravenous injection of graphene and hence evaluation of its hemocompatibility is an essential prerequisite. Herein, both pristine and functionalized graphene are extensively characterized for their interactions with murine macrophage RAW 264.7 cells and human primary blood components. Detailed analyses of the potential uptake by macrophages, effects on its metabolic activity, membrane integrity, induction of reactive oxygen stress, hemolysis, platelet activation, platelet aggregation, coagulation cascade, cytokine induction, immune cell activation, and immune cell suppression are performed using optimized protocols for nanotoxicity evaluation. Electron microscopy, confocal Raman spectral mapping, and confocal fluorescence imaging studies show active interaction of both the graphene systems with macrophage cells, and the reactive oxygen species mediated toxicity effects of hydrophobic pristine samples are significantly reduced by surface functionalization. In the case of hemocompatibility, both types of graphene show excellent compatibility with red blood cells, platelets, and plasma coagulation pathways, and minimal alteration in the cytokine expression by human peripheral blood mononuclear cells. Further, both samples do not cause any premature immune cell activation or suppression up to a relatively high concentration of 75 μg mL(-1) after 72 h of incubation under in vitro conditions. This study clearly suggests that the observed toxicity effects of pristine graphene towards macrophage cells can be easily averted by surface functionalization and both the systems show excellent hemocompatibility.
石墨烯及其衍生物被提议用于几种重要的生物医学应用，包括药物输送、基因输送、对比成像和抗癌治疗。这些应用大多数都需要静脉内注射石墨烯，因此评估其血液相容性是一个必要的前提条件。在这里，我们对原始和功能化的石墨烯进行了广泛的表征，以研究它们与小鼠巨噬细胞 RAW 264.7 细胞和人原代血液成分的相互作用。通过优化的纳米毒性评估方案，对巨噬细胞的潜在摄取、对其代谢活性、膜完整性、诱导活性氧应激、溶血、血小板激活、血小板聚集、凝血级联、细胞因子诱导、免疫细胞激活和免疫细胞抑制的影响进行了详细分析。电子显微镜、共聚焦拉曼光谱映射和共聚焦荧光成像研究表明，两种石墨烯系统都与巨噬细胞发生了积极的相互作用，并且疏水性原始样品的活性氧物种介导的毒性作用通过表面功能化得到了显著降低。在血液相容性方面，两种类型的石墨烯都与红细胞、血小板和血浆凝血途径具有极好的相容性，并且对人外周血单核细胞的细胞因子表达的改变很小。此外，在体外条件下孵育 72 小时后，两种样品在相对较高的浓度 75 μg mL(-1)下，不会引起任何过早的免疫细胞激活或抑制。这项研究清楚地表明，通过表面功能化可以很容易地避免原始石墨烯对巨噬细胞的毒性作用，并且这两种系统都表现出优异的血液相容性。
