EYE DAMAGE REVERSIBILITY IN AN IN VITRO MODEL OF BOVINE CORNEA TO REPLACE THE DRAIZE TEST COMPLETELY.

BENEDETTI, MARTINA; LENZE, MARIELA; ROCO, JULIETA; MARTÍNEZ, ROMINA; MAGAÑA GUERRERO, FÁTIMA SOFIA ; GARFIAS, YONATHAN ; GUTIÉRREZ, MARÍA LAURA

Niagara Falls

Congreso; 12th World Congress on Alternatives and Animal Use in the Life Sciences; 2023

Alternative Congress Trust

One of the requirements for chemical product registration such as agrochemicals, is toprovide evidence about their potential eye damage. The Draize test performed in rabbitsallows the products to be classified into four categories, considering both the severity ofthe lesions produced in the animal´s eye as well as its healing time. The availablealternative methods to this live animal test do not allow documenting the damagereversibility, nor the time necessary for such reversibility to occur, as required by theGHS classifications. Our proposal is to complement the in vitro model that uses thebovine cornea as a substrate to predict whether a substance is irritating or non-irritating(BCOP), with a strategy that allows predicting if the observed irritation is reversible andthe time it takes to revert.We isolated bovine limbal-stem cells, known to play an important repairing role incorneal injury, and used them as model in a cytotoxicity assay to evaluate the cellsensitivity to reference products. Moreover, a wound healing assay was performed tostudy whether these products differentially affect the replication and migration capacityof the cells. Furthermore, a tissue explant and an organotypic cornea culture model wereimplemented to study if the chemical exposure alters cell’s replication, migration andoverall wound healing differentially.The results obtained from the cytotoxicity test, the wound healing assay and the explantculture show that these methods are effective to improve prediction to the fourcategories. Also, damage reversibility was observed in the organotypic cornea culturemodel.