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; GUTIÉRREZ, MARÍA LAURA

Berlin

Congreso; Microphysiological Systems World Summit; 2023

The International MPS Society

One of the requirements for the registration of chemicals 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. 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. Also, a wound healing assay was 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. A combination of the approaches used have been proven effective to detect the four categories of GHS reference products. This project complements the BCOP in vitro model with a strategy that aims to predict if the observed irritation is reversible which is necessary to finally replace the Draize test completely.