Biocompatibility of the vital dye Acid Violet-17 on retinal pigment epithelial cells

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