Pristine (C60) and hydroxylated [C60(OH)24] fullerene phototoxicity towards HaCaT keratinocytes: type I vs type II mechanisms

Chem Res Toxicol. 2008 May;21(5):1056-63. doi: 10.1021/tx800056w. Epub 2008 Apr 19.

Abstract

The increasing use of fullerene nanomaterials has prompted widespread concern over their biological effects. Herein, we have studied the phototoxicity of gamma-cyclodextrin bicapped pristine C 60 [(gamma-CyD) 2/C 60] and its water-soluble derivative C 60(OH) 24 toward human keratinocytes. Our results demonstrated that irradiation of (gamma-CyD) 2/C 60 or C 60(OH) 24 in D 2O generated singlet oxygen with quantum yields of 0.76 and 0.08, respectively. Irradiation (>400 nm) of C 60(OH) 24 generated superoxide as detected by the EPR spin trapping technique; superoxide generation was enhanced by addition of the electron donor nicotinamide adenine dinucleotide (reduced) (NADH). During the irradiation of (gamma-CyD) 2/C 60, superoxide was generated only in the presence of NADH. Cell viability measurements demonstrated that (gamma-CyD) 2/C 60 was about 60 times more phototoxic to human keratinocytes than C 60(OH) 24. UVA irradiation of human keratinocytes in the presence of (gamma-CyD) 2/C 60 resulted in a significant rise in intracellular protein-derived peroxides, suggesting a type II mechanism for phototoxicity. UVA irradiation of human keratinocytes in the presence of C 60(OH) 24 produced diffuse intracellular fluorescence when the hydrogen peroxide probe Peroxyfluor-1 was present, suggesting a type I mechanism. Our results clearly show that the phototoxicity induced by (gamma-CyD) 2/C 60 is mainly mediated by singlet oxygen with a minor contribution from superoxide, while C 60(OH) 24 phototoxicity is mainly due to superoxide.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Cell Line
  • Cell Survival
  • Free Radicals / chemistry
  • Free Radicals / metabolism
  • Fullerenes / chemistry
  • Fullerenes / metabolism
  • Fullerenes / toxicity*
  • Humans
  • Hydrogen Peroxide / metabolism
  • Hydroxylation
  • Keratinocytes / cytology*
  • Keratinocytes / drug effects*
  • Keratinocytes / metabolism
  • Molecular Structure
  • Oxygen / metabolism
  • Photochemistry
  • Spectrophotometry

Substances

  • Free Radicals
  • Fullerenes
  • pristine (C60)
  • Hydrogen Peroxide
  • Oxygen