An Assay System to Evaluate Riboflavin/UV-A Corneal Phototherapy Efficacy in a Porcine Corneal Organ Culture Model

SIMPLE SUMMARY: The scope of this study is to quantitatively evaluate, with an automated digital image analysis method, the efficacy of riboflavin/UV-A corneal phototherapy on the cornea in a porcine corneal organ culture model of ulcerative melting keratitis. Riboflavin/UV-A corneal phototherapy provided a favorable outcome in the corneal wound healing process after chemical injury: the treatment restores the damaged corneas to the texture of healthy corneas. This automated image analysis method may be compared to clinical diagnostic methods, such as optical coherence tomography (OCT) imaging, for in vivo damaged ocular structural investigations. Positive results from this research could provide an opportunity for studying the effects of this method in other economically and emotionally valued species, such as dogs, cats, and horses. The relatively overall low treatment cost and the ease of performing the procedure make riboflavin/UV-A corneal phototherapy accessible to the veterinary market. ABSTRACT: The purpose of this study was to investigate the response of porcine corneal organ cultures to riboflavin/UV-A phototherapy in the injury healing of induced lesions. A porcine corneal organ culture model was established. Corneal alterations in the stroma were evaluated using an assay system, based on an automated image analysis method able to (i) localize the holes and gaps within the stroma and (ii) measure the brightness values in these patches. The analysis has been performed by dividing the corneal section in 24 regions of interest (ROIs) and integrating the data analysis with a “multi-aspect approach.” Three group of corneas were analyzed: healthy, injured, and injured-and-treated. Our study revealed a significant effect of the riboflavin/UV-A phototherapy in the injury healing of porcine corneas after induced lesions. The injured corneas had significant differences of brightness values in comparison to treated (p < 0.00) and healthy (p < 0.001) corneas, whereas the treated and healthy corneas showed no significant difference (p = 0.995). Riboflavin/UV-A phototherapy shows a significant effect in restoring the brightness values of damaged corneas to the values of healthy corneas, suggesting treatment restores the injury healing of corneas after lesions. Our assay system may be compared to clinical diagnostic methods, such as optical coherence tomography (OCT) imaging, for in vivo damaged ocular structure investigations.