Trypan blue: effect on retinal pigment epithelial and neurosensory retinal cells.
OBJECTIVE
To evaluate the toxicity of trypan blue on retinal cells in vitro.


METHODS
Human retinal pigment epithelial cells (ARPE-19) and rat neurosensory retinal cells (R28) were grown in tissue culture and treated with four different concentrations of trypan blue (0.1%, 0.05%, 0.025%, and 0.0125%), in combination with surgical light exposure (0, 5, or 10 minutes). Cell viability, mitochondrial function, and DNA synthesis were measured by trypan blue dye-exclusion assay, mitochondrial dehydrogenase assay, and tritiated [3H] thymidine incorporation, respectively.


RESULTS
ARPE-19 and R28 cells exposed to trypan blue with or without illumination did not show any significant decrease, either in cell viability by the dye-exclusion assay or in [3H] thymidine incorporation. R28 cells exposed to 0.1% trypan blue with and without light showed a significant reduction of mitochondrial dehydrogenase activity (P <0.05). ARPE-19 cells exposed to trypan blue, with or without light, did not show any significant decrease in mitochondrial dehydrogenase activity.


CONCLUSIONS
This study suggests that rat neurosensory retina (R28) cells are more sensitive than human RPE (ARPE-19) cells to trypan blue. ARPE-19 cells showed no evidence of toxicity with any of the three assays, but R28 cells showed evidence of toxicity with the mitochondrial dehydrogenase assay at the higher doses and light-exposure times studied. Clinical studies must be conducted to determine the safety and efficacy of staining of the inner limiting membrane with trypan blue.
Narayanan
Raja
R
Department of Ophthalmology, University of California, Irvine, California 92697-4375, USA.
Kenney
M Cristina
MC
Kamjoo
Sami
S
Trinh
Thuan-Hau T
TH
Seigel
Gail M
GM
Resende
Gilberto P
GP
Kuppermann
Baruch D
BD
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004396:Coloring Agents / Q000633:toxicity*; D004247:DNA / Q000096:biosynthesis; D006801:Humans; D008027:Light; D008928:Mitochondria / Q000502:physiology; D010857:Pigment Epithelium of Eye / Q000187:drug effects* / Q000528:radiation effects; D051381:Rats; D012160:Retina / Q000187:drug effects* / Q000528:radiation effects; D012449:Safety; D013385:Succinate Dehydrogenase / Q000378:metabolism; D014343:Trypan Blue / Q000633:toxicity*
D004396:Coloring Agents; D004247:DNA; D013385:Succinate Dehydrogenase; D014343:Trypan Blue