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K. Tokuda, T. Tsukamoto, T. Mita, S. Fujisawa, M. Matsubara; Excitotoxic Neurodegeneration by Vital Stains in Isolated Rat Retinas . Invest. Ophthalmol. Vis. Sci. 2003;44(13):5216.
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Purpose: We evaluated the retinal toxicity of three vital stains (indocyanine green (ICG), trypan blue (TB) and triamcinolone acetonide (TA)), which are widely used to identify the transparent retina and the vitreous body during vitrectomy, to isolated rat retinas using the same experimental method. We firstly compared two kinds of artificial aqueous humors with artificial cerebrospinal fluid (aCSF) before determining the influence of dye. The retinal damage was assessed by morphological examination and biochemical assay measuring the amount of lactate dehydrogenase (LDH) release from the injured cells. Methods: 8-week-old male Sprague-Dawley rats were anesthetized and the eyes were dissected from the orbits. Then the sensory retinal tissue was removed gently from the retinal pigment epithelium and incubated in aCSF for a one-hour recovery period. In experiment 1, retinal pieces were placed in various media, aCSF, BSS plus® (Alcon Laboratories, Ft. Worth, TX, USA), Opeguard® Neo kit (Otsuka, Tokyo, Japan) and PBS for a positive control. In experiment 2, retinal tissues were exposed to one of the three vital stains (ICG, TB and TA) at a clinical concentration for one minute. They were then rinsed with aCSF and incubated in BSS plus® (Alcon) maintained at 37 C° . Samples were taken from each medium every 60 minutes for measurement of LDH. LDH activity was measured spectrophotometrically and statistical analysis was done with repeated measurement ANOVA. Results: In experiment 1, there was no difference in the release of LDH among aCSF, BSS plus® and Opeguard® Neo Kit compared to PBS. Histological findings confirmed the same results. In experiment 2, there were no significant changes in LDH release with the TB and TA solutions. By contrast, tissues exposed to ICG solutions were damaged in every retinal layer and showed significantly higher release of LDH compared to that of the control. Conclusions: Exposure of ICG in the vitreous cavity at a clinical concentration may cause irreversible retinal damage in the human eye.
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