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

Abstract

PurposeTo examine the viability and differentiation of retinal pigment epithelial (RPE) cells after exposure to the vital dye Acid Violet-17 (AV-17).MethodsBovine 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.ResultsExposure 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.ConclusionAV-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.

Toxicity and phototoxicity in human ARPE-19 retinal pigment epithelium cells of dyes commonly used in retinal surgery

“…AV at low concentrations (0.0125%) in the dark was found not to lead to damage of RPE cells isolated from bovine retina; this was, however, different when the concentration was increased to 0.025% (still below a clinically useful concentration). 48 Penha et al 1 had found similar toxicity levels as ours, while Hurst et al 26 found lower values. A recent survey of the literature by Gerding 3 shows that toxic effects of AV are found at concentrations well below those that were used clinically.…”

Safety Profile of Lutein- Versus Triamcinolone Acetonide–Based Vitreous Staining

“… 26 Consistently, ARPE-19 cells have been largely used to study retinal cytotoxicity and the biocompatibility of vital dyes. 27 – 31 Finally, the reliability of the results obtained using ARPE-19 cells has been supported by several previous studies evaluating the retinal cytotoxicity/biocompatibility of various intraocular medical devices. For example, comparable results have been documented when retinal ganglion cells and ARPE-19 cells have been used to assess cell viability after exposure to silicone oil (SO), 32 as well as ARPE-19 cell and human retina ex vivo culture model to assess cell viability following exposure to non-cytotoxic PFCL samples.…”

Vital Dyes in Vitreomacular Surgery

“…Preclinical evaluation suggested lack of toxicity at concentrations up to 0.125 mg/ml, 119,120 and some concern for phototoxicity. 92 Ala Medics introduced Ala Purple to the market at a concentration of 1.5 mg/ml.…”