title: Oxidative stress and inflammatory response in dermal toxicity of single-walled carbon nanotubes.
abstract: Single-walled carbon nanotubes (SWCNT) represent a novel material with unique electronic and mechanical properties. The extremely small size ( approximately 1 nm diameter) renders their chemical and physical properties unique. A variety of different techniques are available for the production of SWCNT; however, the most common is via the disproportionation of gaseous carbon molecules supported on catalytic iron particles (high-pressure CO conversion, HiPCO). The physical nature of SWCNT may lead to dermal penetration following deposition on exposed skin. This dermal deposition provides a route of exposure which is important to consider when evaluating SWCNT toxicity. The dermal effects of SWCNT are largely unknown. We hypothesize that SWCNT may be toxic to the skin. We further hypothesize that SWCNT toxicity may be dependent upon the metal (particularly iron) content of SWCNT via the metal's ability to interact with the skin, initiate oxidative stress, and induce redox-sensitive transcription factors thereby affecting/leading to inflammation. To test this hypothesis, the effects of SWCNT were assessed both in vitro and in vivo using EpiDerm FT engineered skin, murine epidermal cells (JB6 P+), and immune-competent hairless SKH-1 mice. Engineered skin exposed to SWCNT showed increased epidermal thickness and accumulation and activation of dermal fibroblasts which resulted in increased collagen as well as release of pro-inflammatory cytokines. Exposure of JB6 P+ cells to unpurified SWCNT (30% iron) resulted in the production of ESR detectable hydroxyl radicals and caused a significant dose-dependent activation of AP-1. No significant changes in AP-1 activation were detected when partially purified SWCNT (0.23% iron) were introduced to the cells. However, NFkappaB was activated in a dose-dependent fashion by exposure to both unpurified and partially purified SWCNT. Topical exposure of SKH-1 mice (5 days, with daily doses of 40 microg/mouse, 80 microg/mouse, or 160 microug/mouse) to unpurified SWCNT caused oxidative stress, depletion of glutathione, oxidation of protein thiols and carbonyls, elevated myeloperoxidase activity, an increase of dermal cell numbers, and skin thickening resulting from the accumulation of polymorphonuclear leukocytes (PMNs) and mast cells. Altogether, these data indicated that topical exposure to unpurified SWCNT, induced free radical generation, oxidative stress, and inflammation, thus causing dermal toxicity.
authors.affiliation: Health Effects Laboratory Division, Pathology and Physiology Research Branch, NIOSH, M/L 2015, 1095 Willowdale Road, Morgantown, WV 26505, United States. zsk1@cdc.gov
authors.forename: A R
authors.initials: AR
authors.lastname: Murray
authors.forename: E
authors.initials: E
authors.lastname: Kisin
authors.forename: S S
authors.initials: SS
authors.lastname: Leonard
authors.forename: S H
authors.initials: SH
authors.lastname: Young
authors.forename: C
authors.initials: C
authors.lastname: Kommineni
authors.forename: V E
authors.initials: VE
authors.lastname: Kagan
authors.forename: V
authors.initials: V
authors.lastname: Castranova
authors.forename: A A
authors.initials: AA
authors.lastname: Shvedova
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003094:Collagen / Q000378:metabolism; D016207:Cytokines / Q000096:biosynthesis; D004578:Electron Spin Resonance Spectroscopy; D005609:Free Radicals / Q000276:immunology; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D007249:Inflammation / Q000139:chemically induced*; D051379:Mice; D008812:Mice, Hairless; D016328:NF-kappa B / Q000096:biosynthesis / Q000235:genetics; D037742:Nanotubes, Carbon / Q000633:toxicity*; D010078:Oxazines; D018384:Oxidative Stress / Q000187:drug effects*; D009195:Peroxidase / Q000378:metabolism; D012867:Skin / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D012871:Skin Diseases / Q000139:chemically induced* / Q000473:pathology; D023822:Tissue Engineering; D018808:Transcription Factor AP-1 / Q000096:biosynthesis / Q000235:genetics; D014966:Xanthenes
chemical_list: D016207:Cytokines; D005609:Free Radicals; D016328:NF-kappa B; D037742:Nanotubes, Carbon; D010078:Oxazines; D018808:Transcription Factor AP-1; D014966:Xanthenes; C005843:resazurin; D003094:Collagen; D009195:Peroxidase; D005978:Glutathione