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Kai Januschowski, Sebastian Mueller, Martin Spitzer, Johanna Hofmann, Charlotte Schramm, Maximilian Schultheiß, Karl-Ulrich Barth-Schmidt, Peter Szurman; Testing The Biocompatibility Of Two New Heavy Intraocular Dyes For Vitreoretinal Surgery Using An Isolated Perfused Vertebrate Retina Organ Culture Model And Retinal Ganglion Cells. Invest. Ophthalmol. Vis. Sci. 2011;52(14):5670.
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During vitreoretinal surgery vital dyes are used to visualize anatomical structures, e.g for the removal of epiretinal membranes (ERM). The rate of reoccurrence is negatively influenced by remaining parts of the ERM on the inner limiting Membrane (ILM) using it as a scaffold. To facilitate staining of the inner limiting membrane without preceding fluid-air exchange, substances with a density higher than water are added to generate a heavy dye. Brilliant Blue (BBG) with 4% polyethyleneglycol (PEG) and BBG with Trypan Blue (TB) and 4% PEG are two new dyes. It was the purpose of this study to evaluate biocompatibilty using retinal ganglion cells and an isolated superfused retinal pseudo in-vivo model for electrophysiological evaluation.
To determine cytotoxicity of ILM Blue and MBB Dual for 30, 60,1 20 and 320 seconds monolayer cultures of retinal ganglion cells (RGC5) were used. For functionality testing bovine retinas were isolated and superfused with an oxygen saturated nutrient solution, and the electroretinogram (ERG) was recorded. The two dyes were applied epiretinally for 30, 60 and 120 seconds. ERG-recovery was monitored for 75 minutes.
After staining with BBG with 4% PEG and BBG with TB and 4% PEG no statistical significant reduction of a- or b-wave amplitudes were recorded at the end of the washout except for a 29.79% decrease (p=0.0043) of a wave amplitudes after 30 sec of staining with BBG with TB and 4% PEG. Only immediately after staining with BBG with TB and 4% PEG for 60 seconds a 60.78% (p=0.0088) reduction of b-wave amplitudes was recorded and a significant (p=0.0085) 42.00% reduction of a-wave amplitudes immediately after staining for 120 seconds. During the MTT assay we noted not significant difference in cell viability after 30,60,120 and 320 seconds of staining with BBG with 4% PEG or BBG with TB and 4% PEG in comparison to the control group (DMEM) after formazan extraction.
The two heavy dyes BBG with 4% PEG and BBG with TB and 4% PEG seem to be safe for the clinical use for a staining perior of up to 120 seconds, probably even up to 320 seconds.
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