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Majid Rouhbakhshzaeri, Behrad Yousefi Milani, Elham Ghahari, Sanaz Gidfar, Ali R Djalilian; Designing a reproducible model of corneal endothelial injury by phacoemulsification in pig eyes. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):5734.
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© ARVO (1962-2015); The Authors (2016-present)
We need a reliable and repeatable model of corneal endothelial injury for many further investigations. We tried different methods to establish this injury model and standardize it.
Fresh pig eye globes were obtained. In the first technique, we excised the cornea with the scleral rim and tried several methods of direct mechanical and phacoemulsification injury. An Alcon Legacy 20000 machine using a phacoemulsification handpiece was used with varying degrees of ultrasound power and duration in a constant distant from corneas in cell culture plates. In the second method, the ultrasound injury was induced in intact globes following a clear cornea incision with the chamber maintained by irrigation of balanced salt solution. After the injury, the corneas were excied and stained with Trypan Blue and Alizarin Red and the proportion of dead/live cells was calculated.
Direct mechanical injury on an excised cornea with a defined object resulted in a measurable injury but this was felt not be applicable to in vivo application. Injury on excised cornea with phacoemulsification/ultrasound produced variable degrees of injury since the cornea was mobile in the plate and when it was fixed it still had buldging forward/backward. Phacoemulsification/ultrasound injury in the anterior chamber in whole globes provided the most repeatable degree of injury as measured by the proportion of dead/live endothelial cells.
Phacoemulsification/ultrasound injury in intact pig globes can be used as a measurable and repeatable injury model compatible with clinical scenarios. We can use this model later to investigate the protective effect of treatments on corneal endothelial cells.
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