PURPOSE: To explore the use of an ex vivo, in situ porcine cornea as a model for dry eye (exposure keratitis). METHODS: Twenty-seven porcine eyes were obtained from freshly killed animals at the local abattoir. The viability of 9 corneas (control-baseline group) was assessed within 5 minutes after enucleation on site. A further 18 eyes were transported to the laboratory, where they were exposed to ambient conditions for 4 hours (experimental group A, 6 eyes), for 6 hours (experimental group B, 6 eyes), and for 4 hours with wetting with Dulbecco Phosphate-Buffered Saline every 5 minutes (exposure control group, 6 eyes). All corneas were assessed by trypan blue exclusion for cell viability. RESULTS: The number of dead cells in the central region was significantly greater than those in the peripheral region (P < 0.05) in all groups. The number of dead cells in both corneal areas increased significantly (P < 0.05) in the experimental groups with time of exposure, but there was no significant increase in the exposure control group. CONCLUSION: Preliminary data on the number of dead cells in porcine corneal epithelium after enucleation and the effect of exposure were obtained. It was found that after exposure to air, the corneal cells were maintained well by regular wetting, but there was progressively greater cell damage with exposure without wetting. These baseline data will be useful for the further development of the porcine dry eye model to investigate exposure keratitis.
PURPOSE: To explore the use of an ex vivo, in situ porcine cornea as a model for dry eye (exposure keratitis). METHODS: Twenty-seven porcine eyes were obtained from freshly killed animals at the local abattoir. The viability of 9 corneas (control-baseline group) was assessed within 5 minutes after enucleation on site. A further 18 eyes were transported to the laboratory, where they were exposed to ambient conditions for 4 hours (experimental group A, 6 eyes), for 6 hours (experimental group B, 6 eyes), and for 4 hours with wetting with Dulbecco Phosphate-Buffered Saline every 5 minutes (exposure control group, 6 eyes). All corneas were assessed by trypan blue exclusion for cell viability. RESULTS: The number of dead cells in the central region was significantly greater than those in the peripheral region (P < 0.05) in all groups. The number of dead cells in both corneal areas increased significantly (P < 0.05) in the experimental groups with time of exposure, but there was no significant increase in the exposure control group. CONCLUSION: Preliminary data on the number of dead cells in porcine corneal epithelium after enucleation and the effect of exposure were obtained. It was found that after exposure to air, the corneal cells were maintained well by regular wetting, but there was progressively greater cell damage with exposure without wetting. These baseline data will be useful for the further development of the porcine dry eye model to investigate exposure keratitis.
Authors: Dinesh Jaishankar; Jason S Buhrman; Tibor Valyi-Nagy; Richard A Gemeinhart; Deepak Shukla Journal: Invest Ophthalmol Vis Sci Date: 2016-01-01 Impact factor: 4.799