Peruzynska Magdalena, Cendrowski Krzysztof, Barylak Martyna, Tkacz Marta, Piotrowska Katarzyna, Kurzawski Mateusz, Mijowska Ewa, Drozdzik Marek
Department of Experimental & Clinical Pharmacology, Pomeranian Medical University,Powstancow Wlkp. 72, 70-111 Szczecin, Poland.
Institute of Chemical and Environment Engineering, West Pomeranian University of Technology, Pulaskiego 10, 70-322 Szczecin, Poland.
Toxicol In Vitro. 2017 Jun;41:205-213. doi: 10.1016/j.tiv.2017.03.005. Epub 2017 Mar 18.
In recent years, graphene and its derivatives have been extensively investigated because of their unique properties, which can be used in many fields including biomedical applications. Therefore, detailed biological study is required. In the current paper the detailed toxicological studies on single and four layer graphene oxide (GO) nanoflakes is presented. The morphology and size of the nanomaterials were characterized via atomic force microscopy. Cytotoxicity, proliferation and internalization study were performed using various methods, including optical, confocal and Raman microscopy imaging, flow cytometry analysis, colorimetric and luminescent cell assays. Our first findings undeniably show that the nanomaterials' functionalization has a considerable impact on their behavior in a biological environment. The cytotoxicity assay confirmed comparable, dose dependent cytotoxicity of single and four layers GO flakes. The differences between these two nanomaterials became more distinct during cell proliferation study and ROS detection. Namely, markedly stronger inhibition of cell proliferation and higher ROS generation by one-layer GO-PEG than four-layer GO-PEG were observed. Cell imaging revealed efficient internalization of the both GO nanoflakes in a time dependent manner. These findings emphasize the role of number of layer and functionalization in GO toxicological characteristics and may provide helpful information for their further biomedical applications.
近年来，石墨烯及其衍生物因其独特的性质而受到广泛研究，这些性质可应用于包括生物医学应用在内的许多领域。因此，需要进行详细的生物学研究。在当前论文中，展示了对单层和四层氧化石墨烯（GO）纳米片的详细毒理学研究。通过原子力显微镜对纳米材料的形态和尺寸进行了表征。使用各种方法进行了细胞毒性、增殖和内化研究，包括光学、共聚焦和拉曼显微镜成像、流式细胞术分析、比色法和发光细胞测定。我们的初步研究结果无可否认地表明，纳米材料的功能化对其在生物环境中的行为有相当大的影响。细胞毒性测定证实了单层和四层GO薄片具有可比的剂量依赖性细胞毒性。在细胞增殖研究和活性氧检测过程中，这两种纳米材料之间的差异变得更加明显。具体而言，观察到单层GO-PEG对细胞增殖的抑制作用明显强于四层GO-PEG，且活性氧生成量更高。细胞成像显示两种GO纳米片均以时间依赖性方式有效内化。这些发现强调了层数和功能化在GO毒理学特性中的作用，并可能为其进一步的生物医学应用提供有用信息。
