Indocyanine green increases light-induced oxidative stress, senescence, and matrix metalloproteinases 1 and 3 in human RPE cells.
OBJECTIVE
Indocyanine green (ICG) is a commonly used vital dye for macular surgery. Recent reports implicate that its use might be associated with less favourable results regarding postoperative visual outcome and damage of retinal cells, and atrophic degeneration of the retinal pigment epithelium (RPE) has been described. This study investigates the effects of ICG on light-induced senescence of RPE cells.


METHODS
Primary human RPE cells were either pre-incubated with ICG in concentrations of 0.005% and 0.05% or not and then exposed to white light. After 10 min of irradiation viability, induction of intracellular reactive oxygen species (ROS) and senescence-associated β-galactosidase activity (SA β-Gal) were determined. Expression and secretion of matrix metalloproteinases (MMPs) 1 and 3 and their mRNA were determined by RT-PCR and ELISA.


RESULTS
Light exposure decreased RPE cell viability by 46%. Treatment with 0.005% and 0.05% ICG alone decreased RPE cell viability by 7% and 21%. In addition, expression of ROS, SA β-Gal, and MMP-1 and 3 was significantly increased. When 0.005% and 0.05% ICG treatments were combined with light exposure, viability decreased by 69% and 82% compared to the untreated control. Effects on the expression of ROS, SA β-Gal, and MMP-1 and 3 were, depending on the ICG dose, significantly increased when cells were pre-incubated with ICG and then illuminated.


CONCLUSIONS
In this study, pretreatment with ICG significantly increased light-induced oxidative stress and senescence. This might indicate a potential, supplementary mechanism that could explain RPE alterations and reduced functional results after ICG-assisted internal limiting membrane peeling.
Kernt
Marcus
M
Department of Ophthalmology, Ludwig Maximilian University, Munich, Germany. marcus.kernt@med.uni-muenchen.de
Hirneiss
Christoph
C
Wolf
Armin
A
Liegl
Raffael
R
Rueping
Johann
J
Neubauer
Aljoscha
A
Alge
Claudia
C
Ulbig
Michael
M
Gandorfer
Arndt
A
Kampik
Anselm
A
Haritoglou
Christos
C
D002478:Cells, Cultured; D016922:Cellular Senescence / Q000528:radiation effects*; D004396:Coloring Agents / Q000633:toxicity*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007208:Indocyanine Green / Q000633:toxicity*; D008027:Light; D020781:Matrix Metalloproteinase 1 / Q000235:genetics / Q000378:metabolism*; D019278:Matrix Metalloproteinase 3 / Q000235:genetics / Q000378:metabolism*; D018384:Oxidative Stress / Q000528:radiation effects*; D012333:RNA, Messenger / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D055213:Retinal Pigment Epithelium / Q000187:drug effects* / Q000201:enzymology / Q000528:radiation effects; D001616:beta-Galactosidase / Q000378:metabolism
D004396:Coloring Agents; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; C061901:GLB1 protein, human; D001616:beta-Galactosidase; D019278:Matrix Metalloproteinase 3; D020781:Matrix Metalloproteinase 1; D007208:Indocyanine Green