Grantee Research Project Results
2005 Progress Report: Evaluating Nanoparticle Interactions with Skin
EPA Grant Number: R831715Title: Evaluating Nanoparticle Interactions with Skin
Investigators: Monteiro-Riviere, Nancy A. , Riviere, Jim E.
Institution: North Carolina State University
EPA Project Officer: Hahn, Intaek
Project Period: September 18, 2004 through September 30, 2007
Project Period Covered by this Report: September 18, 2004 through September 30, 2005
Project Amount: $328,972
RFA: Exploratory Research to Anticipate Future Environmental Issues: Impacts of Manufactured Nanomaterials on Human Health and the Environment (2003) RFA Text | Recipients Lists
Research Category: Safer Chemicals , Nanotechnology , Human Health
Objective:
The focus of this research project is to assess the nature of interaction between manufactured nanoparticles and the skin including dermal absorption, and cutaneous toxicity as well as the ability of these materials to distribute to the skin after systemic exposure. These studies will use iron oxide nanocrystals, cadmium selenide nanocrystals, and carbon fullerene nanoparticles that are representative of the broad spectrum of nanoparticles presently being used by industry. Eight particle types selected from these commercially relevant manufactured nanoparticles will be studied to allow assessment of size, shape, and composition on absorption, distribution, or toxicity to the skin.
All studies will be conducted in three well-characterized in vitro skin models: human skin keratinocyte cell cultures; porcine skin flow-through diffusion cells; and the isolated perfused porcine skin flap (IPPSF). Nanoparticles will be applied topically in three different exposure scenarios at different doses to assess potential dermal absorption in the diffusion cell studies and to assess cellular toxicity (transmission electron microscopy, viability) and irritation (IL-8 release) in cell culture. Those particles that penetrate skin or cause direct irritation then will be completely characterized in IPPSF studies, which have previously been shown to be predictive of in vivo absorption in humans. Similarly, to model nanoparticle uptake into skin after systemic exposure, nanoparticles will be infused into the arterial blood supply of the IPPSF to assess the ability to distribute out of the vasculature into the skin. Deposition of particles in epidermal tissue after both infusion and topical exposure will be evaluated using high-resolution electron microscopy.
Progress Summary:
Dermal exposure is a significant route of exposure for nanomaterials. Little is known, however, about the toxicity of nanomaterials, and no information is available on their effects on protein expression in keratinocytes. Multiwall carbon nanotubes (MWCNT), fullerenes, and fullerene-based amino acids (BAA), and quantum dots (QD) were studied in human epidermal keratinocytes. The biological effects of these nanomaterials were studied using transmission electron microscopy, atomic force microscopy, confocal microscopy, viability tests, and cytokine assays. The interactions of nanomaterials in cultured human keratinocytes, absorption of nanomaterials into skin, and confocal images depicting localization within skin were evaluated; MWCNT were primarily localized in intracytoplasmic vacuoles and induced the release of the proinflammatory cytokine interleukin 8 (IL-8) in a time dependent manner. In addition, a two-dimensional electrophoresis-based proteomic approach identified several proteins that were altered significantly by MWCNT. These data clearly show that MWCNT, not derivatized nor optimized for biological applications, are capable of intracellular localization within human epidermal keratinocytes (HEK) and irritation in a target cell that composes a primary route of occupational exposure for manufactured nanotubes. Studies also were conducted with the fullerenes nanoC60 and derivatized C60 (C60(OH)24) in HEK. In addition, fullerene BAA and fullerene C60 peptide with a fluorescently tagged nuclear localization signal (NLS-FITC [fluorescein isothiocyanate]) for 24 hours and 48 hours were assessed in HEK. We have evaluated QD of two different sizes and three different surface coatings (polyethylene glycol, carboxylic acid, or amine) in flow through diffusion cells. QD of different sizes, shapes, and surface coatings can penetrate intact skin at an occupationally relevant dose within the span of an average-length workday.
The permeability of skin to nanomaterials of different sizes, surface coatings, or in different vehicles indicates that the skin may be permeable to nanomaterials with diverse physicochemical properties. These findings are important for risk assessment. Our results conducted in HEK and in-flow through diffusion cells assessed the cellular uptake, penetration, cytotoxicity, and inflammatory potential of several types of nanomaterials. Furthermore these findings strongly indicate that intact skin could be an important route of localized, and possibly systemic, exposure of humans to engineered nanoscale materials.
Future Activities:
We will continue to conduct our research on fullerenes, nanoC60 and derivatized C60 (C60(OH)24) in human epidermal keratinocytes, and study their effects with surfactants. Also, we will continue to study the effects of fullerene BAA and fullerene C60 peptide with a NLS-FITC in HEK, as well as QD of two different sizes and three different surface coatings (polyethylene glycol, carboxylic acid, or amine).
These ongoing studies will be conducted in HEK to assess cellular uptake, cytotoxicity, and inflammatory potential. The fullerene NLS-FITC and QD will be studied in porcine flow through diffusion cells to assess skin penetration by confocal microscopy.
Journal Articles on this Report : 5 Displayed | Download in RIS Format
Other project views: | All 119 publications | 22 publications in selected types | All 17 journal articles |
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Holsapple MP, Farland WH, Landry TD, Monteiro-Riviere NA, Carter JM, Walker NJ, Thomas KV. Research strategies for safety evaluation of nanomaterials, Part II: toxicological and safety evaluation of nanomaterials, current challenges and data needs. Toxicological Sciences 2005;88(1):12-17. |
R831715 (2005) R831712 (Final) |
Exit Exit Exit |
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Monteiro-Riviere NA, Inman AO, Wang YY, Nemanich RJ. Surfactant effects on carbon nanotube interactions with human keratinocytes. Nanomedicine: Nanotechnology, Biology, and Medicine 2005;1(4):293-299. |
R831715 (2005) R831715 (2006) R831710 (Final) |
Exit |
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Monteiro-Riviere NA, Nemanich RJ, Inman AO, Wang YY, Riviere JE. Multi-walled carbon nanotube interactions with human epidermal keratinocytes. Toxicology Letters 2005;155(3):377-384. (Most highly cited paper by Elsevier publishers.). |
R831715 (2005) R831715 (2006) |
Exit Exit Exit |
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Monteiro-Riviere NA, Inman AO. Challenges for assessing carbon nanomaterial toxicity to the skin. Carbon 2006;44(6):1070-1078. |
R831715 (2005) R831715 (2006) R831712 (Final) |
Exit |
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Oberdorster G, Maynard A, Donaldson K, Castranova V, Fitzpatrick J, Ausman K, Carter J, Karn B, Kreyling W, Lai D, Olin S, Monteiro-Riviere N, Warheit D, Yang H. Principles for characterizing the potential human health effects from exposure to nanomaterials: elements of a screening strategy. Particle and Fibr Toxicology 2005;2(8):1-35. |
R831715 (2005) R831715 (2006) |
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Supplemental Keywords:
nanoparticles, nanotubes, quantum dots, nanomaterials, surfactants, skin absorption, skin toxicity, skin exposure, dermal absorption, cell culture, health effects, risk assessment, cellular toxicity, transmission electron microscopy, diffusion cells, human epidermal keratinocytes,, Scientific Discipline, Health, ENVIRONMENTAL MANAGEMENT, Environmental Chemistry, Health Risk Assessment, Risk Assessments, Biology, Risk Assessment, toxicology, carbon fullerene, cutaneous toxicity, nanotechnology, iron oxide nanocrystals, dermal contact, human exposure, cellular response to nanoparticles, exposure assessment, human health risk, oxidative stressProgress and Final Reports:
Original AbstractThe perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.