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Shiri Zayit-Soudry, Michael Mimouni, Marina Chemerovski-Glikman, Yarden Dagan, Esraa Haj, Lihi Adler-Abramovich, Daniel Segal, Ehud Gazit; A method for the selection of cataract disintegrating compounds and their use for reversal of crystalline lens opacification. Invest. Ophthalmol. Vis. Sci. 2016;57(12):2498.
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© ARVO (1962-2015); The Authors (2016-present)
To describe a novel ex vivo assay for examining the efficacy of various compounds for the reversal of human crystalline lens opacification in the search for pharmacological treatment for cataract.
After obtaining informed consent, fragments of cataractous lenses were collected from patients undergoing routine clear corneal incision phacoemulsification cataract surgery. Following quantification of total protein concentration in each cataract sample using the standard Bradford assay, the concentration was adjusted to obtain an optimal absorbance value. Ex vivo cataract samples were then incubated with several concentrations of various compounds being screened for cataract disintegration effect. Reversal of protein aggregates was monitored several times a day up to 48 h using the common turbidity assay. In between measurements samples were kept at 37 °C with constant shaking.
Each compound selected for screening was tested on cataract samples obtained from 5-10 different patients, in triplicates. Initial total protein concentrations ranged between 1-10 mg/mL and after adjustment for optimal absorbance final concentrations ranged between 0.5-3 mg/mL. Of the 15 compounds screened, five showed no effect on protein turbidity measurement over time, while five other showed inconsistent results. Treatment with five compounds resulted in significant reduction in the optical density of the solution, reflecting restoration of transparency of the cataractous samples, two of which showed a dose-dependent effect. Among them each agent demonstrated different efficacy and unique kinetics.
This simple yet innovative experimental approach implements spectrophotometric principles to enable direct testing of the impact of potential compounds on disintegration of actual human crystalline lens material ex vivo. Further biophysical studies, currently conducted by our group, will provide mechanistic insight on the cataract reversal effect.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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