May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
A Comparison of the in vitro Safety of Intraocular Dyes
Author Affiliations & Notes
  • C. M. Hutnik
    Ophthalmology, Ivey Eye Institute, London, Ontario, Canada
  • D. Yuen
    Ophthalmology, Ivey Eye Institute, London, Ontario, Canada
  • H. Liu
    Ophthalmology, Ivey Eye Institute, London, Ontario, Canada
  • A. Proulx
    Ophthalmology, Ivey Eye Institute, London, Ontario, Canada
  • J. Gonder
    Ophthalmology, Ivey Eye Institute, London, Ontario, Canada
  • Footnotes
    Commercial Relationships  C.M. Hutnik, None; D. Yuen, None; H. Liu, None; A. Proulx, None; J. Gonder, None.
  • Footnotes
    Support  Physicians Services Incorporated
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 4884. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      C. M. Hutnik, D. Yuen, H. Liu, A. Proulx, J. Gonder; A Comparison of the in vitro Safety of Intraocular Dyes. Invest. Ophthalmol. Vis. Sci. 2008;49(13):4884.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose: : Another group recently reported that Brilliant blue G (BBG) was a safe dye for for internal limiting membrane (ILM) peeling in vitreoretinal surgery based upon an in vivo animal study. In vitro toxicity wasnot reported.The purpose of the present study was tocompare the in vitro toxicity of BBG to indocyanine green (ICG), the current gold standard, as well as to trypan blue (TB) and evans blue (EB). A demonstration of in vitro safety would further support a potential paradigm shift in the first choice of adjuvant dye used for ILM peeling.

Methods: : The toxicity of four dyes was tested in two different cell cultures. These were a commercially available human retinal pigment epithelial cell line known as ARPE-19 and a primary mixed murine retinal ganglion/ muller glial cell culture (RGC). The dose-dependent toxicity of the dyes was determined by exposing the two different cell cultures to each dye, at four different concentrations, for a short exposure time of 3 minutes. In another experiment a medium exposure time of 30 minutes was performed. Cell viability was measured using the MTT assay. Time-dependent toxicity of the dyes was also studied. All four dyes, each diluted to 1/500th of stock concentration, were applied to the ARPE-19 cells for a prolonged exposure, to simulate the possible long lasting persistence of dye in the vitreous cavity after vitreoretinal surgery. Cell viability was measured at the 2, 24, 48 and 72 hour time point using the MTT assay. Finally, the possible influence of osmolarity was assessed.

Results: : After 3 minutes of dye exposure, only BBG caused significant loss of ARPE-19 cell viability. Longer exposure times of 30 minutes resulted in toxicity from both BBG and TB, with the toxicity of BBG being greater than that of TB. Prolonged exposure at 1/500th of stock concentration of either BBG or ICG produced equivalent loss of cell viability. None of the dyes produced any significant loss of RGC/glial cell viability after 3 minutes of exposure. However, 30 minutes of exposure to BBG, TB and EB demonstrated toxicity, with the toxicity of BBG being greater than that of TB and EB. Osmolarity was not a factor in any of the experiments.

Keywords: retinal culture • ocular irritancy/toxicity testing • vitreoretinal surgery 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×