title: Phototoxicity and cytotoxicity of fullerol in human lens epithelial cells.
abstract: The water-soluble, hydroxylated fullerene [fullerol, nano-C60(OH)22-26] has several clinical applications including use as a drug carrier to bypass the blood ocular barriers. We have assessed fullerol's potential ocular toxicity by measuring its cytotoxicity and phototoxicity induced by UVA and visible light in vitro with human lens epithelial cells (HLE B-3). Accumulation of nano-C60(OH)22-26 in the cells was confirmed spectrophotometrically at 405 nm and cell viability estimated using MTS and LDH assays. Fullerol was cytotoxic to HLE B-3 cells maintained in the dark at concentrations higher than 20 microM. Exposure to either UVA or visible light in the presence of >5 microM fullerol-induced phototoxic damage. When cells were pretreated with non-toxic antioxidants: 20 microM lutein, 1 mM N-acetyl cysteine, or 1 mM l-ascorbic acid prior to irradiation, only the singlet oxygen quencher-lutein significantly protected against fullerol photodamage. Apoptosis was observed in lens cells treated with fullerol whether or not the cells were irradiated, in the order UVA>visible light>dark. Dynamic light scattering (DLS) showed that in the presence of the endogenous lens protein alpha-crystallin, large aggregates of fullerol were reduced. In conclusion, fullerol is both cytotoxic and phototoxic to human lens epithelial cells. Although the acute toxicity of water-soluble nano-C60(OH)22-26 is low, these compounds are retained in the body for long periods, raising concern for their chronic toxic effect. Before fullerols are used to deliver drugs to the eye, they should be tested for photo- and cytotoxicity in vivo.
authors.affiliation: Department of Natural Sciences, Fordham University, 113 West 60th Street, New York City, NY 10023, USA. jroberts@fordham.edu
authors.forename: Joan E
authors.initials: JE
authors.lastname: Roberts
authors.forename: Albert R
authors.initials: AR
authors.lastname: Wielgus
authors.forename: William K
authors.initials: WK
authors.lastname: Boyes
authors.forename: Usha
authors.initials: U
authors.lastname: Andley
authors.forename: Colin F
authors.initials: CF
authors.lastname: Chignell
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects*; D017484:Dermatitis, Phototoxic / Q000473:pathology*; D004847:Epithelial Cells / Q000473:pathology* / Q000528:radiation effects; D037741:Fullerenes / Q000633:toxicity*; D006801:Humans; D007202:Indicators and Reagents; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D007908:Lens, Crystalline / Q000473:pathology* / Q000528:radiation effects; D008027:Light; D014975:Lutein / Q000633:toxicity; D009336:Necrosis / Q000473:pathology; D010316:Particle Size; D051381:Rats; D017207:Rats, Sprague-Dawley; D014466:Ultraviolet Rays; D038201:alpha-Crystallins / Q000737:chemistry / Q000187:drug effects
chemical_list: D000975:Antioxidants; D037741:Fullerenes; D007202:Indicators and Reagents; D038201:alpha-Crystallins; C496652:fullerol C60(OH)22; D007770:L-Lactate Dehydrogenase; D053148:Caspase 3; D014975:Lutein