Safety testing of blue vital dyes using cell culture models

J Ocul Pharmacol Ther. 2014 Jun;30(5):406-12. doi: 10.1089/jop.2013.0213. Epub 2014 Feb 7.

Abstract

Purpose: To investigate the safety of trypan blue, brilliant blue G (BBG), Evans blue (EB), patent blue, Chicago blue (CB), and bromophenol blue (BB), with and without halogen and xenon light exposure.

Methods: All dyes were diluted in a balanced saline solution at a concentration of 0.5%. Cells of the human RPE line ARPE-19 and rat RGC5 were exposed to vital dyes for 5 min. Experiments with and without xenon or halogen illumination were performed. The viability of ARPE-19 and RGC5 cells was determined at 12, 24, or 120 h by a cell proliferation assay using WST-1 reagent. The apoptotic events as well as cell numbers were registered for 72 h and counted by time-lapse videomicroscopy.

Results: There was no evidence of ARPE-19 or RGC5 toxicity, immediate (0 and 24 h) or delayed (120 h), following exclusive exposure to each single dye. After halogen light exposure, ARPE-19 cell lines did not show any significant toxicity, except for when they were exposed to EB. After xenon illumination, ARPE-19 cells showed a marked decrease in cell viability when exposed to EB or CB and a moderate decrease when exposed to BBG and BB. After xenon illumination, RGC5 cells showed the highest decrease in cell viability when exposed to EB and CB; BB caused the same decrease in cell viability as in ARPE-19 cells.

Conclusion: Interaction of light from endo-illumination source and blue vital dyes may increase the risk of retinal toxicity.

MeSH terms

  • Animals
  • Benzenesulfonates / adverse effects*
  • Bromphenol Blue / adverse effects*
  • Cell Line, Transformed
  • Cell Survival / drug effects*
  • Cell Survival / physiology
  • Cells, Cultured
  • Coloring Agents / adverse effects
  • Evans Blue / adverse effects*
  • Humans
  • Rats
  • Rosaniline Dyes / adverse effects*
  • Trypan Blue / adverse effects*

Substances

  • Benzenesulfonates
  • Coloring Agents
  • Rosaniline Dyes
  • Bromphenol Blue
  • pontamine sky blue
  • Evans Blue
  • sulfan blue
  • brilliant blue
  • Trypan Blue