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Milestone in animal-free testing strategies: two in vitro methods accepted for the first time for the identification of non-irritant chemicals in the field of eye irritation

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Apr 24, 2013

Major contribution to the substitution of the Draize eye irritation test on living rabbits

A major milestone has been achieved in the search of animal-free testing strategies as defined by EURL ECVAM (the European Union Reference Laboratory for Alternatives to Animal Testing) (*):  revised OECD test guidelines extending the applicability domain of two in vitro methods  - the Bovine Corneal Opacity and Permeability (BCOP) test and the Isolated Chicken Eye (ICE) test - have been adopted for the purpose of eye irritation testing.

This is the first time that in vitro methods are accepted for the identification of non-irritant chemicals in the field of eye irritation. Both in vitro methods are using isolated eyes from animals slaughtered for human consumption and will contribute to the substitution of the Draize eye irritation test performed on the eyes of living rabbits.

The revision of two OECD Test Guidelines based on organotypic methods for eye irritation testing was adopted at the OECD meeting of the Working Group of National Coordinators of the Test Guideline Programme held in Paris on 9 to 11 April 2013.

ICEThe two test guidelines had originally been adopted in 2009 for the identification of serious eye damage/eye irritation of chemicals further to retrospective validation by the US Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) in collaboration with EURL ECVAM and the Japanese Center for the Validation of Alternative Methods (JaCVAM). Additional validation of the two tests showed their usefulness also for the identification of chemicals not requiring classification for serious eye damage/eye irritation (i.e. non-irritant chemicals), thus leading to the revision of the test guidelines under the co-lead of EURL ECVAM and the Netherlands.

(*) Scott, L., Eskes, C., Hoffman, S., Adriaens, E., Alepee, N., Bufo, M., Clothier, R., Facchini, D., Faller, C., Guest, R., Hamernik, K., Harbell, J., Hartung, T., Kamp, H., Le Varlet, B., Meloni, M., Mcnamee, P., Osborn, R., Pape, W., Pfannenbecker, U., Prinsen, M., Seaman, C., Spielmann H., Stokes, W., Trouba, K., Vassallo, M., Van den Berghe, C., Van Goethem, F., Vinardell, P. & Zuang, V. (2010). Bottom-up and Top-down Approach: Eye Irritation Testing Strategy to Reduce and Replace in vivo studies. Toxicology In Vitro 24, 1-10.


a) BCOP method: rinsing the bovine cornea. Photo reproduced courtesy of the Institute of In Vitro Science (USA);

b) ICE method: chicken eye opacity evaluated via slit-lamp microscope. Photo reproduced courtesy of TNO (The Netherlands).