The Effects of Vital Dyes on Retinal Pigment Epithelium Cells in Oxidative Stress

Abstract

Purpose: To determine the effect of the most commonly used vital dyes in vitrectomy [trypan blue at 0.15% concentration and indocyanine green (ICG) at 0.5% concentration] on the viability of human retinal pigment epithelium (RPE) cell lines (ARPE-19) exposed to oxidative stress. Methods: ARPE-19 cells unexposed or exposed to oxidative stress (hypoxic chamber) were treated for 1 min with one of the dyes. RPE proliferation was measured by ³H-thymidine incorporation, adhesion by ability to adhere to fibronectin, and safety by annexin V staining. Results: Proliferation: The dyes affected the proliferation of RPE cells differently under non-hypoxic and hypoxic conditions (p = 0.001). In non-hypoxic conditions, there was no statistically significant difference between the proliferation of the treated (both dyes) and untreated RPE cells (p = 0.279). Under hypoxia, both dyes significantly suppressed proliferation, more so with ICG (p = 0.001). Adhesion: The dyes affected adhesion differently under non-hypoxic and hypoxic conditions (p = 0.04). In non-hypoxic conditions, both increased the adhesive properties of RPE cells to fibronectin, ICG more than trypan blue (p = 0.001). Under hypoxia, both dyes suppressed adhesion, with no statistically significant difference between treated and non-treated RPE cells. Apoptosis: Both dyes increased early apoptosis of RPE cells compared with no treatment (p = 0.001), ICG more than trypan blue. Hypoxia increased the apoptosis of both dyes compared to non-hypoxic conditions (p = 0.02). Conclusions: In hypoxic conditions, both dyes showed an inhibition of RPE adhesion to fibronectin and proliferation capacity and an increase in early apoptosis compared with non-hypoxic conditions. Apoptosis was greater in ICG-treated RPE cells than in trypan blue-treated cells.

Insulin‐like growth factor‐1 activates PI3K/Akt signalling to protect human retinal pigment epithelial cells from amiodarone‐induced oxidative injury

“…The presence of IGF‐1 in the supernatant of cultured RPE cells and in the interphotoreceptor matrix suggests that RPE‐derived IGF is active on photoreceptor cells (Holtkamp et al, ). ROS‐induced oxidative stress increases in RPE cells with ageing and decreased cell density causing age‐related macular degeneration (Golan et al, ). IGF‐1 promotes the survival of the cells under oxidative stress (Ayadi et al, ).…”

Membrane Blue Dual Protects Retinal Pigment Epithelium Cells/Ganglion Cells—Like through Modulation of Mitochondria Function

“…Regarding TB, ARPE-19 cells showed no evidence of toxicity, unlike the rat neurosensory retinaR28 cells whose mitochondrial dehydrogenase activity was found to be damaged at the higher doses/light-exposure times [ 37 ]. Furthermore, in non-hypoxic conditions, TB increased the adhesive properties of RPE cells to fibronectin, whereas under hypoxia, it suppressed adhesion, reduced proliferation, and increased apoptosis [ 38 ].…”

Intravenous Injection of Indocyanine Green Results in an Artificial Transient Desaturation by Pulse Oximetry

“…Both dyes decreased rates of proliferation and adhesion to fibronectin, as well as increased apoptosis when placed under hypoxic conditions. 11 As apoptosis is a controlled cell-directed process, it may result in more oxygen availability to the remaining viable tissue.…”