Clinical Ophthalmology (Jul 2016)
Biocompatibility of the vital dye Acid Violet-17 on retinal pigment epithelial cells
Abstract
Ayşegül Tura,1 Aizhan Alt,1 Julia Lüke,1 Salvatore Grisanti,1 Christos Haritoglou,2 Carsten H Meyer,3 Khaled Nassar,1 Matthias Lüke1 On behalf of the International Chromovitrectomy Collaboration 1Department of Ophthalmology, University of Schleswig-Holstein, Lübeck, Germany; 2Department of Ophthalmology, Ludwig-Maximilians University, Munich, Germany; 3Department of Ophthalmology, Pallas Clinic, Aarau, Switzerland Purpose: To examine the viability and differentiation of retinal pigment epithelial (RPE) cells after exposure to the vital dye Acid Violet-17 (AV-17). Methods: Bovine RPE cells were incubated with AV-17 (0.0625–0.5 mg/mL) for 30 seconds or 5 minutes. Viability was determined by live/dead staining, cleaved CASP3 immunostainings, and MTT test. Actin cytoskeleton was visualized by Alexa 488-phalloidin. Immunocytochemistry was performed to determine the levels of ZO-1, CTNNB1, and KRT19. Results: Exposure to AV-17 at the concentrations of 0.25–0.5 mg/mL resulted in a dose-dependent decrease in viability, the loss of ZO-1 from tight junctions, translocation of CTNNB1 into the cytoplasm and nucleus, disarrangement of the actin cytoskeleton, and a slight increase in KRT19. Conclusion: AV-17 at a concentration <0.125 mg/mL is likely to be well tolerated by the RPE cells, whereas the concentrations from 0.25 mg/mL onward can reduce viability and induce dedifferentiation particularly after long-term exposure. Keywords: Acid Violet-17, retinal pigment epithelial cells, biocompatibility, viability, differentiation
