May 2003
Volume 44, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2003
Safety Testing of Indocyanine Green (ICG) and Trypan Blue Using a Cell Culture Model
Author Affiliations & Notes
  • T.L. Jackson
    Ophthalmology, Rayne Institute, London, United Kingdom
  • J. Zhang
    Ophthalmology, Rayne Institute, London, United Kingdom
  • J. Hillenkamp
    Ophthalmology, Rayne Institute, London, United Kingdom
  • A.R. El-Osta
    Ophthalmology, Rayne Institute, London, United Kingdom
  • J. Marshall
    Ophthalmology, Rayne Institute, London, United Kingdom
  • Footnotes
    Commercial Relationships  T.L. Jackson, None; J. Zhang, None; J. Hillenkamp, None; A.R. El-Osta, None; J. Marshall, None.
  • Footnotes
    Support  Special Trustees of Guys and St Thomas' Hospital, Allerton Trust
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 4461. doi:
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      T.L. Jackson, J. Zhang, J. Hillenkamp, A.R. El-Osta, J. Marshall; Safety Testing of Indocyanine Green (ICG) and Trypan Blue Using a Cell Culture Model . Invest. Ophthalmol. Vis. Sci. 2003;44(13):4461.

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

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Abstract

Abstract: : Purpose: To undertake safety testing of indocyanine green (ICG) and trypan blue using a cell culture model. Methods: A human retinal pigment epithelium (RPE) cell-line was incubated with ICG over a range of concentrations up to 0.5% and trypan blue up to 0.2%. Cells were exposed to the agent for 5, 15 or 30 minutes, rinsed, and incubated 24 hours. Cell viability was measured using a mitochondrial dehydrogenase-assay and fluorescent live-dead probe. Experiments were repeated using ICG 0.5% and 1%, and trypan blue 0.06% and 0.12%, with follow-up at 0,1,5 and 15 days. ICG experiments were repeated in the presence of illumination from a surgical endolight Results: Using the enzyme-assay, no data point fell below 2SD of the negative control. There was no clear relationship between cell viability and the concentration of the agent or duration of follow-up, except in cells exposed to 1% ICG. These showed a linear (R2 0.9952) decline in viability with time with a significant reduction by day 15 (p 0.016). Cells incubated with ICG and endoillumination were not significantly different from those incubated with ICG and no endoillumination (p 0.263), illumination only (p 0.238), or the negative control (p 0.615). The live-dead probe did not demonstrate any evidence of cell damage. Conclusions: ICG and trypan blue have been advocated as vital stains for use during macular surgery. Concentrations of up to 0.5% ICG and 0.2% trypan blue do not measurably reduce the viability of cultured RPE cells. The suggestion of concentration dependent delayed toxicity with 1% ICG indicates that the lowest efficacious concentration should be used for vital staining.

Keywords: retina • retinal pigment epithelium • macular holes 
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