Eye Damage Reversibility in an in Vitro Model of Bovine Cornea to Replace the Draize Test Completely

BENEDETTI, MARTINA; LENZE, MARIELA; ROCO, JULIETA; GUTIÉRREZ, MARÍA LAURA

Montevideo

Congreso; SETAC Latin America 15th Biennial Meeting; 2023

SETAC

One of the requirements for the registration of substances such as agrochemicals is to provide evidence about their potential eye damage. The Draize test performed in rabbits allows the products to be classified into four categories, considering both the severity of the lesions produced in the animal's eye as well as its healing time. The available alternative methods to this live animal test do not allow documenting the damage reversibility, nor the time necessary for such reversibility to occur, as required by the UN GHS classifications.Our proposal is to complement the in vitro model that uses the bovine 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 and the time it takes to revert. This is necessary to finally replace the Draize test completely.Limbal stem cells are known to play an important repairing role in corneal injury; therefore we isolated these cells from bovine cornea and used them to evaluate the cell sensitivity to reference products. A wound healing assay was also performed to study whether these products differentially affect the replication and migration capacity of the cells. Furthermore, a tissue explant and an organotypic cornea culture model were implemented to study if the chemical exposure alters cell’s replication, migration and overall wound healing differentially.In conclusion, a combination of the approaches used have been proven effective to detect the fourcategories of GHS reference products