Biochemical and Biophysical Research Communications
ROS and NF-κB are involved in upregulation of IL-8 in A549 cells exposed to multi-walled carbon nanotubes
Section snippets
Materials and methods
Materials. Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum, and F12 were products of Gibco BRL (Grand Island, NY, USA). NF-κB consensus oligonucleotide was purchased from Promega (Madison, WI, USA). γ-[32P]ATP was the product of NEN Life Science (Boston, MA, USA). Antibodies against p65, IκBa, PARP, β-actin, and secondary antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Escherichia coli 0111:B4 lipopolysaccharide (LPS) was obtained from Sigma
Cytotoxicity, cell size and complexity analysis
Among the widely used formazan-based cytotoxicity assays, WST-1 has been shown not to interact with CNTs, in contrast to 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) [21]. Therefore, cytotoxicity induced by MWCNTs was assessed with the WST-1 assay. After 72 h incubation with A549 and BEAS-2A cells, MWCNTs elicited a concentration-dependent toxic effect on cell viability (Fig. 1A). The cell viability results were confirmed by a live–dead cell assay, which indicated that
Discussion
In the present study, we evaluated the toxicological potential of MWCNTs on pulmonary epithelial cells by studying their effects on cell viability, cell size and complexity, induction of oxidative stress, release of inflammatory mediators, and activation of NF-κB, which led to a number of interesting findings.
MWCNTs appeared to be water-miscible and dispersed at the concentration range used in the presence of the Pluronic surfactant as a dispersing agent, not toxic by itself (data not shown).
Acknowledgments
Project supported by the Science and Technology Innovation Project of Fujian Province for Young Scientific Researchers, China (2008F3097) and the Special Program for Key Basic Research of the Ministry of Science and Technology, China (2006CB93300).
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