Evaluation of silver nanoparticle toxicity in skin in vivo and keratinocytes in vitro

Environ Health Perspect. 2010 Mar;118(3):407-13. doi: 10.1289/ehp.0901398. Epub 2009 Oct 23.

Abstract

Introduction: Products using the antimicrobial properties of silver nanoparticles (Ag-nps) may be found in health and consumer products that routinely contact skin.

Objectives: This study was designed to assess the potential cytotoxicity of Ag-nps in human epidermal keratinocytes (HEKs) and their inflammatory and penetrating potential into porcine skin in vivo.

Materials and methods: We used eight different Ag-nps in this study [unwashed/uncoated (20, 50, and 80 nm particle diameter), washed/uncoated (20, 50, and 80 nm), and carbon-coated (25 and 35 nm)]. Skin was dosed topically for 14 consecutive days. HEK viability was assessed by MTT, alamarBlue (aB), and CellTiter 96 AQueous One (96AQ). Release of the proinflammatory mediators interleukin (IL)-1beta, IL-6, IL-8, IL-10, and tumor necrosis factor-alpha (TNF-alpha) were measured.

Results: The effect of the unwashed Ag-nps on HEK viability after a 24-hr exposure indicated a significant dose-dependent decrease (p < 0.05) at 0.34 microg/mL with aB and 96AQ and at 1.7 microg/mL with MTT. However, both the washed Ag-nps and carbon-coated Ag-nps showed no significant decrease in viability at any concentration assessed by any of the three assays. For each of the unwashed Ag-nps, we noted a significant increase (p < 0.05) in IL-1beta, IL-6, IL-8, and TNF-alpha concentrations. We observed localization of all Ag-nps in cytoplasmic vacuoles of HEKs. Macroscopic observations showed no gross irritation in porcine skin, whereas microscopic and ultrastructural observations showed areas of focal inflammation and localization of Ag-nps on the surface and in the upper stratum corneum layers of the skin.

Conclusion: This study provides a better understanding Ag-nps safety in vitro as well as in vivo and a basis for occupational and risk assessment. Ag-nps are nontoxic when dosed in washed Ag-nps solutions or carbon coated.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cell Survival / drug effects
  • Cells, Cultured
  • Dose-Response Relationship, Drug
  • Evaluation Studies as Topic
  • Humans
  • Inflammation / chemically induced
  • Inflammation / metabolism
  • Inflammation / pathology
  • Interleukins / metabolism
  • Keratinocytes / cytology
  • Keratinocytes / drug effects*
  • Keratinocytes / metabolism
  • Metal Nanoparticles / administration & dosage
  • Metal Nanoparticles / toxicity*
  • Particle Size
  • Silver / administration & dosage
  • Silver / toxicity*
  • Skin / cytology
  • Skin / drug effects*
  • Skin / metabolism
  • Swine
  • Time Factors
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • Interleukins
  • Tumor Necrosis Factor-alpha
  • Silver