title: Effect of vital dyes on retinal pigmented epithelial cell viability and apoptosis: implications for chromovitrectomy.
abstract: OBJECTIVE
  To investigate the in vitro effect of vital dyes on toxicity and apoptosis in a human retinal pigment epithelial cell line.
  
  
  METHODS
  ARPE-19 cells were exposed to brilliant blue (BBG), Evans Blue (EB), bromophenol blue (BroB), indocyanine green (ICG), infracyanine green (IfCG), light green (LG), fast green (FG), indigo carmine (IC) and Congo red (CR). Balanced salt solution was used as the control. Five different concentrations and 2 exposure times were tested. Cell viability was determined by the MTS (1-solution methyl thiazolyl tetrazolium) assay and apoptosis by Bax expression on Western blot.
  
  
  RESULTS
  All dyes significantly reduced cell viability after 3 min of exposure at all concentrations (p < 0.01), except for BBG that was safe at concentrations up to 0.25 mg/ml and CR up to 0.05 mg/ml, while LG was safe at all concentrations. Toxicity was higher after 30 min of exposure. Expression of Bax was upregulated after all dye exposures, except BBG; ICG had the highest Bax expression (p < 0.01).
  
  
  CONCLUSIONS
  Overall the safest dye was BBG followed by LG, IfCG, FG, CR, IC, BroB, EB and ICG. ICG was toxic at all concentrations and exposure times tested. Moreover, BBG was the only dye that did not induce apoptosis in ARPE-19 cells.
authors.affiliation: Departamento de Oftalmologia, Instituto da Visão IPEPO, Universidade Federal de São Paulo, São Paulo, Brazil.
authors.forename: Fernando M
authors.initials: FM
authors.lastname: Penha
authors.forename: Marianne
authors.initials: M
authors.lastname: Pons
authors.forename: Elaine de Paula Fiod
authors.initials: Ede P
authors.lastname: Costa
authors.forename: Eduardo B
authors.initials: EB
authors.lastname: Rodrigues
authors.forename: Mauricio
authors.initials: M
authors.lastname: Maia
authors.forename: Maria E
authors.initials: ME
authors.lastname: Marin-Castaño
authors.forename: Michel Eid
authors.initials: ME
authors.lastname: Farah
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004396:Coloring Agents / Q000633:toxicity*; D006801:Humans; D055213:Retinal Pigment Epithelium / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D013997:Time Factors; D014821:Vitrectomy; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D004396:Coloring Agents; D051028:bcl-2-Associated X Protein
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