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The wide application of carbon nanomaterials in various fields urges in-depth understanding of the toxic effects and underlying mechanisms of these materials on biological systems. Cell autophagy was recently recognized as an important lysosome-based pathway of cell death, and autophagosome accumulation has been found to be associated with the exposure of various nanoparticles, but the underlying mechanisms are still uncertain due to the fact that autophagosome accumulation can result from autophagy induction and/or autophagy blockade. In this study, we first evaluated the toxicity of acid-functionalized single-walled carbon nanotubes and graphene oxides, and found that both carbon nanomaterials induced adverse effects in murine peritoneal macrophages, and GOs were more potent than AF-SWCNTs. Both carbon nanomaterials induced autophagosome accumulation and the conversion of LC3-I to LC3-II. However, degradation of the autophagic substrate p62 protein was also inhibited by both nanomaterials. Further analyses on lysosomes revealed that both carbon nanomaterials accumulated in macrophage lysosomes, leading to lysosome membrane destabilization, which indicates reduced autophagic degradation. The effects of AF-SWCNTs and GOs on cell autophagy revealed by this study may shed light on the potential toxic mechanism and suggest caution on their utilization., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

It is increasingly important to understand the single-walled carbon nanotubes' (SWCNTs) immune response as their increasingly biomedical researches and applications. Macrophages and T cells play important roles in scavenging foreign materials and pathogens and regulating immune response. In this work, primarily cultured murine peritoneal macrophages and purified splenic T cells were utilised to determine the toxic effects of SWCNTs and acid-functionalised SWCNTs (AF-SWCNTs) on the immune system, especially on macrophage functions. Macrophages were exposed to 0-50 μg/ml of CNTs for 24 h and no significant cytotoxicity was found by live/dead and annexin-V-FITC/PI analyses. The TEM images revealed that AF-SWCNTs were engulfed mostly through phagocytosis and located in lysosomes of macrophages. Measurement of mitochondrial membrane potential and proteasome subunit gene expression demonstrated that 10 and 50 μg/ml AF-SWCNTs could damage mitochondrial function and proteasome formation in a concentration-dependent manner. Functional analyses revealed that the percentage of phagocytic cells were affected significantly by 20 μg/ml CNTs, and 5 μg/ml AF-SWCNTs inhibited the phagocytic efficiency of latex beads in macrophages. The accessory cell function was affected by both AF-SWCNTs and SWCNTs at concentrations of 10 and 50 μg/ml, respectively. Furthermore, AF-SWCNT biased naïve T-cell differentiation to Th1 type by inducing the production of IFN-γ and TNF, implying the potential risk of Th1-associated diseases (e.g. autoimmune diseases and inflammation) on AF-SWCNT exposure.

Single-walled carbon nanotubes (SWCNTs) are widely used in industrial and medical sectors, and the increasing exposure of SWCNTs necessitates the studies of their potential environmental and health effects. Considerable efforts have been made to improve the dispersion of SWCNTs by chemical modifications. However, the toxicological effects of such modifications on SWCNTs are mostly unknown. This study was designed to determine the influences of acid functionalization on SWCNT toxicity and to understand the molecular toxic mechanisms. RAW264.7 cells were exposed to 0-50 μg/mL of as-synthesized SWCNTs or acid-functionalized SWCNTs (AF-SWCNTs) for 24 hours and then their toxicities were compared via viability analysis. After that the global gene expression profiles of cells exposed to AF-SWCNTs were obtained and analyzed. The results showed that AF-SWCNTs penetrated cell membrane and aggregated in cell cytoplasm and nuclear areas, resulting in enhanced toxicity. In addition, AF-SWCNTs altered the expression of genes related to ribosome, mitochondria, inflammatory response, cell cycle/apoptosis, and proteasome pathway. The gene expression study excluded the interference of metallic impurities and suggested similar toxic mechanism to that of ultra-fine particulate matters.