title: Toxic effects of indocyanine green, infracyanine green, and trypan blue on the human retinal pigmented epithelium.
abstract: BACKGROUND
  Indocyanine green, infracyanine green, and trypan blue are frequently used as aids to visualize structures removed during vitreoretinal surgery. But they may have toxic effects on the retina. We therefore compared the acute and chronic toxicities of these stains on cultured human retinal pigmented epithelial (RPE) cells using clinically relevant concentrations and an identical experimental setup for each agent.
  
  
  METHODS
  Monolayers of RPE cells were incubated with various concentrations of indocyanine green, infracyanine green (each at 0.005%, 0.05%, and 0.5%) or trypan blue (0.05%, 0.06%, 0.1%, 0.15%, and 0.5%) for 5 min (acute exposure) or 6 days (chronic exposure). Using the propidium iodide assay, acute cytotoxicity was monitored at 15-min intervals for up to 3 h. Chronic cytotoxicity was assessed by monitoring cell calcein esterase activity, cell proliferation, and cell morphology (viability) after 6 days of exposure.
  
  
  RESULTS
  Indocyanine and infracyanine green induced acute and chronic toxicities at a concentration above 0.05%. Trypan blue evoked no acute toxicity, but it was chronically cytotoxic at all tested concentrations.
  
  
  CONCLUSIONS
  Despite thorough rinsing after application, significant amounts of the not sufficiently water soluble indocyanine and infracyanine green are retained after surgery by the eye. Trypan blue, being more water-soluble than ICG, is probably retained to the least degree. This circumstance is fortunate given that trypan blue exhibits a chronic cytotoxicity comparable to ICG at all clinically relevant concentrations. During vitrectomy, surgeons should aim to expose retinal tissue to only low concentrations of these stains and for as short a period as possible.
authors.affiliation: Department of Ophthalmology, Inselspital, University of Bern, 3010 Bern, Switzerland.
authors.forename: L
authors.initials: L
authors.lastname: Kodjikian
authors.forename: T
authors.initials: T
authors.lastname: Richter
authors.forename: M
authors.initials: M
authors.lastname: Halberstadt
authors.forename: F
authors.initials: F
authors.lastname: Beby
authors.forename: F
authors.initials: F
authors.lastname: Flueckiger
authors.forename: M
authors.initials: M
authors.lastname: Boehnke
authors.forename: J G
authors.initials: JG
authors.lastname: Garweg
mesh_terms: D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D004396:Coloring Agents / Q000633:toxicity*; D006801:Humans; D007208:Indocyanine Green / Q000031:analogs & derivatives* / Q000633:toxicity*; D010857:Pigment Epithelium of Eye / Q000187:drug effects*; D014343:Trypan Blue / Q000633:toxicity*
chemical_list: D004396:Coloring Agents; C116320:Infracyanine green; D014343:Trypan Blue; D007208:Indocyanine Green