Defensive effects of fullerene-C60/liposome complex against UVA-induced intracellular reactive oxygen species generation and cell death in human skin keratinocytes HaCaT, associated with intracellular uptake and extracellular excretion of fullerene-C60.
The UVA-irradiation of 10 J/cm(2) on HaCaT keratinocytes increased 59.1% of the intracellular reactive oxygen species (ROS) by NBT assay and the cell viability decreased to 31.5% by WST-1 assay, comparing to the non-irradiated control. In the presence of fullerene-C60 (C60) incorporated in phospholipid membrane vehicle (LiposomeFullerene: Lpsm-Flln) of 250-500 ppm, they were restored to -9.1% to +2.3% of the ROS and 83.0-84.8% of the cell viability, but scarcely restored by the liposome without C60 (Lpsm). In HaCaT cells administered with Lpsm-Flln (150 ppm), C60 was ingested at the intracellular concentrations of 1.4-21.9 ppm for 4-24 h, and, intracellular C60 was excreted by 80% at 4h after rinsing-out, and decreased to 2-10% after 24-48 h. C60 was predominantly distributed around the outside of nuclear membrane without deterioration of intact cell morphology according to fluorescent immunostain. Thus Lpsm-Flln is found to be an effective antioxidant that could preserve HaCaT keratinocytes against UVA-induced cellular injury. Lpsm-Flln has a potential to serve as a cosmetic material for skin protection against UVA.
Kato
Shinya
S
Laboratory of Cell-Death Control BioTechnology, Faculty of Life and Environmental Sciences, Prefectural University of Hiroshima, 562, Nanatsuka, Shobara, Hiroshima 727-0023, Japan.
Kikuchi
Risa
R
Aoshima
Hisae
H
Saitoh
Yasukazu
Y
Miwa
Nobuhiko
N
D017209:Apoptosis; D002460:Cell Line; D037741:Fullerenes / Q000008:administration & dosage* / Q000737:chemistry; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism / Q000528:radiation effects; D008081:Liposomes / Q000737:chemistry*; D011837:Radiation-Protective Agents / Q000008:administration & dosage* / Q000737:chemistry; D017382:Reactive Oxygen Species / Q000378:metabolism*; D014466:Ultraviolet Rays
D037741:Fullerenes; D008081:Liposomes; D011837:Radiation-Protective Agents; D017382:Reactive Oxygen Species; C069837:fullerene C60