Induction of cell death by graphene in Arabidopsis thaliana (Columbia ecotype) T87 cell suspensions

J Hazard Mater. 2013 Sep 15;260:1032-41. doi: 10.1016/j.jhazmat.2013.06.063. Epub 2013 Jun 29.

Abstract

The toxicity of graphene on suspensions of Arabidopsis thaliana (Columbia ecotype) T87 cells was investigated by examining the morphology, mitochondrial dysfunction, reactive oxygen species generation (ROS), and translocation of graphene as the toxicological endpoints. The cells were grown in Jouanneau and Péaud-Lenoel (JPL) media and exposed to graphene at concentrations 0-80 mg/L. Morphological changes were observed by scanning electron microscope and the adverse effects such as fragmented nuclei, membrane damage, mitochondrial dysfunction was observed with fluorescence microscopy by staining with Hoechst 33342/propidium iodide and succinate dehydrogenase (mitochondrial bioenergetic enzyme). Analysis of intracellular ROS by 2',7'-dichlorofluorescein diacetate demonstrated that graphene induced a 3.3-fold increase in ROS, suggesting that ROS are key mediators in the cell death signaling pathway. Transmission electron microscopy verified the translocation of graphene into cells and an endocytosis-like structure was observed which suggested graphene entering into the cells by endocytosis. In conclusion, our results show that graphene induced cell death in T87 cells through mitochondrial damage mediated by ROS.

Keywords: Arabidopsis thaliana; Endocytosis; Mitochondrial dysfunction; Necrosis; ROS.

MeSH terms

  • Apoptosis / drug effects*
  • Arabidopsis / drug effects*
  • Arabidopsis / ultrastructure
  • Benzimidazoles / chemistry
  • Cell Membrane / drug effects
  • Cell Nucleus / drug effects
  • Cells, Cultured
  • Culture Media / chemistry
  • Dose-Response Relationship, Drug
  • Endocytosis
  • Graphite / chemistry
  • Graphite / toxicity*
  • Membrane Potential, Mitochondrial / drug effects
  • Microscopy, Atomic Force
  • Microscopy, Fluorescence
  • Oxidative Stress
  • Propidium / chemistry
  • Reactive Oxygen Species
  • Succinate Dehydrogenase / chemistry
  • Water / chemistry

Substances

  • Benzimidazoles
  • Culture Media
  • Reactive Oxygen Species
  • Water
  • Propidium
  • Graphite
  • Succinate Dehydrogenase
  • bisbenzimide ethoxide trihydrochloride