Evaluation of short-term bovine eye storage protocol for the enucleated eye toxicity test

The enucleated eye test (EET) has used slaughterhouse eyes to evaluate corneal toxic effects in vitro, but the impact of short-term storage on the EET has not been defined. Selected bovine eyes were used on receipt or stored at room temperature (RT) or 4 degrees C for 6 or 24 hr prior to EET protocol for 4 hr at 37 degrees C; plastic bags or containers were used for storage. Aqueous humour measurements indicated post-mortem acidification. With a bicarbonate-mixed salts solution (BMSS), central corneal thickness (CCT) was maintainable ( +/- 3%) for eyes used on receipt, which showed a 13% increase in CCT after a 5-min challenge with benzalkonium chloride (BAC) 0.016%. The BAC response was attenuated if corneal oedema developed during RT storage in bags, or at 4 degrees C. Storage at RT in individual containers resulted in further deterioration of the aqueous humour and corneal desiccation such that increases in CCT occurred in the EET with both BMSS and BAC challenge. Further studies are needed to define a suitable handling and storage medium.