氧化石墨烯-银纳米复合材料及其原始对应物对巨噬细胞的体外比较毒性
Comparative in vitro toxicity of a graphene oxide-silver nanocomposite and the pristine counterparts toward macrophages.
作者信息
de Luna Luis Augusto Visani, de Moraes Ana Carolina Mazarin, Consonni Sílvio Roberto, Pereira Catarinie Diniz, Cadore Solange, Giorgio Selma, Alves Oswaldo Luiz
机构信息
Laboratory of Solid State Chemistry (LQES), Institute of Chemistry, University of Campinas, Campinas, Brazil.
Laboratory of Leishmaniasis (Lableish), Institute of Biology, University of Campinas, Campinas, Brazil.
出版信息
J Nanobiotechnology. 2016 Feb 24;14:12. doi: 10.1186/s12951-016-0165-1.
BACKGROUND
Graphene oxide (GO) is a highly oxidized graphene form with oxygen functional groups on its surface. GO is an excellent platform to support and stabilize silver nanoparticles (AgNP), which gives rise to the graphene oxide-silver nanoparticle (GOAg) nanocomposite. Understanding how this nanocomposite interacts with cells is a toxicological challenge of great importance for future biomedical applications, and macrophage cells can provide information concerning the biocompatibility of these nanomaterials. The cytotoxicity of the GOAg nanocomposite, pristine GO, and pristine AgNP was compared toward two representative murine macrophages: a tumoral lineage (J774) and peritoneal macrophages collected from Balb/c mouse. The production of reactive oxygen species (ROS) by J774 macrophages was also monitored. We investigated the internalization of nanomaterials by transmission electron microscopy (TEM). The quantification of internalized silver was carried out by inductively coupled plasma mass spectrometry (ICP-MS). Nanomaterial stability in the cell media was investigated overtime by visual observation, inductively coupled plasma optical emission spectrometry (ICP OES), and dynamic light scattering (DLS).
RESULTS
The GOAg nanocomposite was more toxic than pristine GO and pristine AgNP for both macrophages, and it significantly induced more ROS production compared to pristine AgNP. TEM analysis showed that GOAg was internalized by tumoral J774 macrophages. However, macrophages internalized approximately 60 % less GOAg than did pristine AgNP. The images also showed the degradation of nanocomposite inside cells.
CONCLUSIONS
Although the GOAg nanocomposite was less internalized by the macrophage cells, it was more toxic than the pristine counterparts and induced remarkable oxidative stress. Our findings strongly reveal a synergistic toxicity effect of the GOAg nanocomposite. The toxicity and fate of nanocomposites in cells are some of the major concerns in the development of novel biocompatible materials and must be carefully evaluated.
背景
氧化石墨烯（GO）是一种在其表面带有氧官能团的高度氧化的石墨烯形式。GO是用于支撑和稳定银纳米颗粒（AgNP）的优良平台，由此产生了氧化石墨烯 - 银纳米颗粒（GOAg）纳米复合材料。了解这种纳米复合材料如何与细胞相互作用是未来生物医学应用中一项极为重要的毒理学挑战，巨噬细胞能够提供有关这些纳米材料生物相容性的信息。将GOAg纳米复合材料、原始GO和原始AgNP对两种具有代表性的小鼠巨噬细胞的细胞毒性进行了比较：一种是肿瘤谱系（J774），另一种是从Balb/c小鼠收集的腹腔巨噬细胞。还监测了J774巨噬细胞产生活性氧（ROS）的情况。我们通过透射电子显微镜（TEM）研究了纳米材料的内化过程。通过电感耦合等离子体质谱（ICP-MS）对内化银进行定量分析。通过肉眼观察、电感耦合等离子体发射光谱（ICP OES）和动态光散射（DLS）对细胞培养基中纳米材料的稳定性进行了长期研究。
结果
GOAg纳米复合材料对两种巨噬细胞的毒性均高于原始GO和原始AgNP，并且与原始AgNP相比，它显著诱导产生了更多的ROS。TEM分析表明，GOAg被肿瘤性J774巨噬细胞内化。然而，巨噬细胞内化的GOAg比原始AgNP少约60%。图像还显示了细胞内纳米复合材料的降解情况。
结论
尽管GOAg纳米复合材料被巨噬细胞内化的程度较低，但它比原始对应物毒性更大，并诱导了显著的氧化应激。我们的研究结果有力地揭示了GOAg纳米复合材料的协同毒性作用。纳米复合材料在细胞中的毒性和归宿是新型生物相容性材料开发中的一些主要问题，必须仔细评估。
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