title: Comparative effects of six intraocular vital dyes on retinal pigment epithelial cells.
abstract: OBJECTIVE
  To evaluate and compare the effects of the following dyes on human pigmented epithelial cells: indocyanine green (ICG), infracyanine green (IfCG), trypan blue (TB), bromophenol blue (BrB), patent blue (PB), and Brilliant Blue G (BBG).
  
  
  METHODS
  ARPE-19 cells cultured in vitro were exposed to these dyes, and acute and chronic toxicity were evaluated. Cell viability was measured by colorimetry (MTT assay), morphology was observed by phase-contrast microscopy, membrane permeability (CMP) was evaluated by flow cytometry with propidium iodide (PI), and mitochondrial membrane potential (ΔΨm) was measured with 3,3'-dihexyloxacarbocyanine (DiOC(6)(3)).
  
  
  RESULTS
  Each of the studied dyes exhibited toxicity after acute exposure at surgical doses. The presence of light often reduced cell viability, especially when measured 3 hours after incubation in the case of ICG, TB, BrB, and BBG. Morphologic changes were induced by ICG, IfCG, and BBG. Both CMP and ΔΨm were altered after exposure to surgical doses of ICG, TB, PB, and a fourfold surgical dose of BrB. Chronic exposure to residual amounts of some dyes was associated with reduced proliferation and even cell death.
  
  
  CONCLUSIONS
  It appears to be prudent to use the lowest possible dose of each dye, to minimize the risk of toxic effects. This precaution may be particularly important in the case of BrB, which should not be used in excess of 0.5%. In addition, abundant irrigation of the vitreous cavity after surgery to completely remove traces of dye may be of crucial importance, particularly in the case of ICG, in minimizing chronic toxicity.
authors.affiliation: Instituto Clínico-Quirúrgico de Oftalmología, Vizcaya, Spain. morales@icqo.org
authors.forename: María-Celia
authors.initials: MC
authors.lastname: Morales
authors.forename: Vanesa
authors.initials: V
authors.lastname: Freire
authors.forename: Aintzane
authors.initials: A
authors.lastname: Asumendi
authors.forename: Javier
authors.initials: J
authors.lastname: Araiz
authors.forename: Itxaso
authors.initials: I
authors.lastname: Herrera
authors.forename: Gonzalo
authors.initials: G
authors.lastname: Castiella
authors.forename: Iñigo
authors.initials: I
authors.lastname: Corcóstegui
authors.forename: Gonzalo
authors.initials: G
authors.lastname: Corcóstegui
mesh_terms: D001978:Bromphenol Blue / Q000008:administration & dosage / Q000633:toxicity; D002463:Cell Membrane Permeability / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003124:Colorimetry; D004396:Coloring Agents / Q000008:administration & dosage / Q000633:toxicity*; D006801:Humans; D007208:Indocyanine Green / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000633:toxicity; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008858:Microscopy, Phase-Contrast; D055213:Retinal Pigment Epithelium / Q000187:drug effects*; D012394:Rosaniline Dyes / Q000008:administration & dosage / Q000633:toxicity; D014343:Trypan Blue / Q000008:administration & dosage / Q000633:toxicity
chemical_list: D004396:Coloring Agents; C116320:Infracyanine green; D012394:Rosaniline Dyes; D001978:Bromphenol Blue; C008769:sulfan blue; D014343:Trypan Blue; D007208:Indocyanine Green; C004692:coomassie Brilliant Blue