Original articleComparison of the In vitro Safety of Intraocular Dyes Using Two Retinal Cell Lines: A Focus on Brilliant Blue G and Indocyanine Green
Section snippets
Reagents
All cell culture reagents were purchased from Invitrogen Canada Inc (Burlington, Ontario, Canada) unless otherwise indicated. Cell viability assay and all dyes were purchased from Sigma-Aldrich Co (Oakville, Ontario, Canada) unless otherwise indicated.
Retinal Pigment Epithelial Cell Culture
The human retinal pigment epithelial cell line (ARPE-19) was purchased from American Type Culture Collection (Manassas, Virginia, USA). Western blot and immunostaining were done using anti-RPE65 to verify the cell line. ARPE-19 expresses RPE65,
ARPE-19 Cell Cultures
BBG was the only dye to cause significant toxicity after three minutes of exposure. Cell viability differed significantly when compared with control for BBG stock concentration (10 mg/ml, cell viability = 73%) and first dilution (2.5 mg/ml, cell viability = 75%) (Figure 1).
After 30 minutes of exposure, there was no toxicity with ICG and EB. With BBG 30 minute exposure, cell viability differed significantly when compared with control for stock concentration (10 mg/ml, cell viability = 64%),
Discussion
The in vitro cytotoxicity of four dyes was compared in both ARPE-19 and mixed primary RGC/glial cell cultures. This is the first study to examine BBG and EB toxicity under these conditions. Experimental parameters were chosen primarily to mimic intraoperative conditions during macular hole surgery. Short- and medium-exposure times approximated typical intraoperative uses and a prolonged exposure time simulated the possible long-lasting persistence of dilute dye in the vitreous cavity
Darana Yuen, MD, completed medical training at the University of Alberta, Canada. She is currently in her final year of residency in Ophthalmology at the University of Western Ontario. In July 2009, upon completion of residency, she will be pursuing a one year glaucoma fellowship at the University of Toronto under the mentorship of Dr Trope and Dr Buys. Dr Yuen's research interests lie in both clinical and basic science.
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2019, Experimental Eye ResearchCitation Excerpt :Several vital dyes are routinely used during vitreoretinal surgery to stain the internal limiting membrane (ILM). A number of studies have reported the safety of Brilliant Blue-G dye in various in vivo as well as in vitro settings revealing contradictory findings with light as the primary causative factor (Awad et al., 2011; Balaiya et al., 2014b; Giansanti et al., 2014; Jindal et al., 2014; Kawahara et al., 2007; Luke et al., 2005; Mansoor et al., 2015; Penha et al., 2013; Remy et al., 2008; Shukla et al., 2011; Yuen et al., 2009). There are various factors which influence the safety of a dye and cell culture models are very important to decipher the biological mechanisms involved in evaluating them.
Darana Yuen, MD, completed medical training at the University of Alberta, Canada. She is currently in her final year of residency in Ophthalmology at the University of Western Ontario. In July 2009, upon completion of residency, she will be pursuing a one year glaucoma fellowship at the University of Toronto under the mentorship of Dr Trope and Dr Buys. Dr Yuen's research interests lie in both clinical and basic science.
Cindy Hutnik, MD, PhD, is an Associate Professor in the Departments of Ophthalmology and Pathology at the University of Western Ontario, and an Associate Scientist at the Lawson Health Research Institute. Dr Hutnik has a full-time clinical practice at the Ivey Eye Institute in London Ontario and is active in both basic science and clinical research.