Fluorescein leakage test: a useful tool in ocular safety assessment

Toxicology in Vitro : an International Journal Published in Association with BIBRA
M Cottin, A Zanvit

Abstract

The fluorescein leakage test (FLT) provides information on the effects of xenobiotics on the impermeability (gate function) of epithelial cell monolayers, and their recovery after exposure. The aim of this study was to assess the validity of this test in the ocular safety assessment of surfactant-based products with various irritant potencies. Madin-Darby canine kidney cells were grown to confluency on microporous membranes and exposed for 15 min to increasing concentrations of test substances. Damage was evaluated by measuring the amount of Na-fluorescein that passed through the monolayer in 30 min, starting just after exposure. Recovery was assessed 4, 24, 48 and 72 hr later. For each sample and each time point, the amounts of test substance that produced 10% and 20% leakage (FL(10) and FL(20)) compared with a cell-free control were calculated. For the 43 samples, FL(20) values ranged from 0.65 to 1000 mg/ml. These values increased or decreased with time according to the substance. In particular, cell monolayers showed very different recoveries after exposure to anionic and cationic substances with similar initial FL(20) values. These in vitro data correlated well with historical Draize in vivo test data (Spearman's varrho > ...Continue Reading

References

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Citations

Apr 1, 1999·Toxicology in Vitro : an International Journal Published in Association with BIBRA·A ZanvitM Buiatti-Tcheng
Aug 1, 1999·Toxicology in Vitro : an International Journal Published in Association with BIBRA·R ClothierY C Kwong
Aug 19, 2007·Journal of Pharmaceutical Sciences·M P Vinardell, M Mitjans
Nov 30, 2006·The Ocular Surface·John L Ubels, Daniel P Clousing
Apr 28, 2009·Regulatory Toxicology and Pharmacology : RTP·Pauline McNameeJulia Scheel
May 19, 2016·The FEBS Journal·Stine C KlingerMorten S Nielsen
Oct 1, 2005·Alternatives to Laboratory Animals : ATLA·Chantra EskesValérie Zuang
Oct 31, 2002·Alternatives to Laboratory Animals : ATLA·Richard ClothierLinda Noble
Nov 16, 2001·Alternatives to Laboratory Animals : ATLA·P A JonesJ H Fentem
Feb 6, 2021·In Vitro Cellular & Developmental Biology. Animal·Yulia Kaluzhny, Mitchell Klausner
Jun 29, 2018·Chemico-biological Interactions·Artur Christian Garcia da SilvaMarize Campos Valadares

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