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T. L. Gosen, R. Mandiga, S. S. Huang; A Model for the Adult Human Eye Using Trypsin Vitreous Digestion. Invest. Ophthalmol. Vis. Sci. 2010;51(13):6078.
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To identify a model of aging human globes for use in anterior-posterior (AP) acceleration impulse injury to the vitreo-retinal interface (VRI) using fresh porcine globes.
Porcine whole globes shipped overnight on ice from Sioux-Preme Packaging (Sioux City, IA) were used within 24hrs of harvest. A pendulum provided an impulse measured by an Endevco 751-10 accelerometer (10mV/g sensitivity, San Juan Capistrano, CA) in an AP vector to a carriage suspending the globe with non-elastic material placed at the sites of rectus muscle attachment. Globes received an impulse of 500g. Globes then received injections with a 30 gauge needle 2mm from limbus with 0.3cc of trypsin-EDTA (t-E) (Invitrogen Carlsbad, CA) and incubated for 1, 1.5 and 2hour (hr). Treated globes received an impulse of 500 or 100g prior to dissection and inspection for gross VRI injury. One control group was treated but not accelerated and dissected after 6, 18, and 24hr. In another control group, the vitreous was removed and the cavity filled with 0.25% t-E. The controls were assessed for retinal and/or vessel degeneration.
Untreated globes given 500g had no VRI injury (n=10). Globes injected with 0.013% (n=3), 0.025% (n=3) and 0.05% (n=3) t-E incubated 1hr demonstrated 0%, 0%, and 14% VRI injury. Globes injected with 0.25% t-E incubated 1hr (n=8), 1.5hr (n=4), and 2hr (n=4), demonstrated VRI injury in 50%, 75%, and 100%, respectively. Globes injected with 0.25% t-E incubated 2hr and given an impulse of 500g (n=12) or 100g (n=5) revealed 91.7% and 0% injury, respectively. Control globes not accelerated but injected with 0.25% t-E incubated 6hr (n=2), 18hr (n=2), 24hr (n=2) and control globes with vitreous removed, filled with 0.25% t-E and incubated for 0.5hr (n=2) and 1hr (n=2) showed gross retinal or vessel degeneration.
Fresh porcine globes have well formed vitreous analogous to young human eyes, which given 0% damage at 500g with no t-E injection appears to be protective against acceleration VRI injury in the AP vector. The injection of 0.3cc 0.25% t-E and 2hr incubation digests the vitreous and preserves the VRI. This treatment yields an active model for syneretic vitreous, simulating the vitreous of the aging human eye that is vulnerable to VRI damage that could precede rhegmatogenous retinal detachment. The damage gradient in the treated porcine globes at 500g and 100g with 97.1% and 0% VRI injury suggests a definable threshold for deceleration injury exists.
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