Abstract
Purpose :
There is an ongoing debate on the potential toxicity of different vital retinal dyes that are being used to stain the internal limiting membrane (ILM) during vitrectomy. We assessed the cellular toxicity of three different ILM dyes in an in vitro cell viability assay.
Methods :
In vitro viability assays were performed on a human retinal pigment epithelium cell line (ARPE-19) and human Müller glia cell line (MIO-M1). Cell viability was measured using MTS assay following a 15-minute exposure to 50µl of phosphate buffered saline (PBS), Indocyanine Green (ICG), Brilliant Blue G 0.025% (BBG) or Brilliant Blue G 0.025% + 4% polyethylene glycol (BBG-PEG). Cell viability in each group was normalized to the PBS control group. Differences in viability between the different dyes and PBS were assessed using one-way analysis of variance with Bonferroni pairwise comparisons.
Results :
In total, 126 wells were analysed (64 ARPE-19: 18 PBS, 17 ICG, 6 BBG and 23 BBG-PEG; 62 MIO-M1: 18 PBS, 17 ICG, 4 BBG and 23 BBG-PEG). In the ARPE-19 assay, viability after BBG exposure was significantly lowered compared to PBS, BBG-PEG and ICG (P = 0.011, P = 0.010 and P = 0.048, respectively) with no significant differences between PBS, BBG-PEG and ICG (P > 0.05). In the MIO-M1 assay, ICG showed a significantly reduced survival compared to PBS and BBG-PEG (P = 0.002 and P = 0.013, respectively), with no statistically significant differences found between PBS, BBG and BBG-PEG (P > 0.05).
Conclusions :
A 15-minute exposure to BBG and ICG resulted in a significant decreased cellular survival in respectively the ARPE-19 and MIO-M1 viability assay. Exposure to BBG-PEG did not result in a significant decreased cellular survival compared to PBS in either cell line. The composition, purity and concentration of vital dyes may be an important consideration when using these dyes in clinical practice.
This is a 2020 ARVO Annual Meeting abstract.