Comparison of six different methods to assess UVA cytotoxicity on reconstructed epidermis. Relevance of a fluorimetric assay (the calcein-AM) to evaluate the photoprotective effects of alpha-tocopherol

Toxicology in Vitro : an International Journal Published in Association with BIBRA
M S Noel-HudsonJ Wepierre

Abstract

A three-dimensional culture of human keratinocytes exposed at the air-liquid interface has been developed and used in conjunction with fluorimetric, colorimetric and radioligand incorporation assays to assess the in vitro toxicity of UVA. The aims of the study were: (1) to compare the relevance of the neutral red uptake (NR), MTT metabolism, (35)S-methionine incorporation, IL1-alpha release and calcein-AM esterification assays for the evaluation of UVA injury; (2) to test the preventive protective effect of an emulsion containing 3% of tocopherol applied on the reconstructed epidermis, in comparison with an application of tocopherol 3% diluted in culture medium either on the apical compartment or in the underneath compartment of the skin culture insert. Viability measurement methods are based on different endpoints. None of the five endpoints measured produced LD(50) values (40 J/cm(2)) that differed significantly from the others. However, calcein-AM assay was relatively more reproducible and easier to handle than the others, and seemed to be a better choice for the evaluation of the protective effects of the tocopherol emulsion. Tocopherol diluted in culture medium under the epidermis 24 hr before irradiation failed to protect...Continue Reading

References

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Citations

Aug 24, 1999·Journal of Cellular Physiology·N Chouinard, M Rouabhia
Jun 15, 1999·Medical & Biological Engineering & Computing·O DamourA F Black
Jul 31, 2013·Journal of Photochemistry and Photobiology. B, Biology·Julie A Evans-JohnsonC-Y Oliver Chen
Feb 24, 2001·Antioxidants & Redox Signaling·M H Ben-MahdiC Pasquier
May 26, 2005·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Daniela BratosinJean Montreuil
Sep 25, 2010·Journal of Gerontological Social Work·Joan K Davitt, Lenard W Kaye

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