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

Abstract

Although recent data highlight the greater protective effects exerted by Membrane Blue Dual (MBD), a precise analysis of the mechanisms of action is missing. We examined the effects of MBD with/without polyethylene glycol (PEG) on both human retinal pigment epithelial cells (ARPE-19) and retinal ganglion cells-like (RGC-5) cultured in the presence/absence of ultraviolet B (UVB) treatment on mitochondria function, oxidants, and apoptosis. In ARPE-19/RGC-5 cells either treated or not with UVB, the effects of MBD with/without PEG were evaluated by specific assays for viability, mitochondrial membrane potential and mitochondrial reactive oxygen species (mitoROS) release. Annexin V was used to detect apoptosis, whereas trypan blue and the scratch assay were used for proliferation/migration. In both physiologic conditions and in the presence of UVB, MBD with/without PEG increased cell viability, mitochondrial membrane potential, proliferation and migration in both ARPE-19 and RGC-5 cells. In general, the effects of MBD with PEG were greater than those caused by MBD without PEG. Our results suggest that, in particular, MBD with PEG is a safe and effective dye for vitreoretinal surgery through the modulation of mitochondrial function.

Plasma Pattern of Extracellular Vesicles Isolated from Hepatitis C Virus Patients and Their Effects on Human Vascular Endothelial Cells

“…These data about the harmful effects of EVs from non-healthy subjects are in agreement with those obtained by other groups which demonstrated that the EVs isolated from patients with various diseases—such as atherosclerosis, acute coronary syndrome, or chronic inflammatory diseases—can induce endothelial dysfunction in various in vitro and ex vivo models [ 59 , 60 ], probably due to common pathophysiological mechanisms including impaired vasorelaxation and induction of vascular inflammation through increased levels of adhesion molecules, ROS, and proinflammatory cytokines [ 61 ]. The use of the MTT and JC-1 assay, which are widely adopted to examine cell viability and mitochondrial membrane potential [ 62 ], allowed us to highlight the aforementioned harmful effects of EVs. Indeed, both cell viability and mitochondrial membrane potential decreased in HUVEC.…”

“…Mitochondrial membrane potential (ΔψM) is an important parameter of the mitochondrial function used as an indicator of cell health, as previously tested in similar cellular models [ 61 , 62 , 63 , 64 , 65 , 66 , 67 , 68 , 69 , 70 , 71 ].…”

Effects of Acetyl-L-Carnitine on Oxidative Stress in Amyotrophic Lateral Sclerosis Patients: Evaluation on Plasma Markers and Members of the Neurovascular Unit

“…In HUVEC and astrocytes, the viability was examined using the 1% 3-[4,5-dimethylthiazol -2-yl]-2,5-diphenyl tetrazolium bromide (MTT; Life Technologies Italia, Monza, Italy) dye [27,34,35]. To perform this analysis, 50,000 HUVEC/astrocytes/well were plated in 24-Transwells plates in complete medium (DMEM supplemented with 10% FBS).…”

“…We used the JC-1 assay in order to examine the mitochondrial membrane potential of HUVEC/astrocytes, as was the case in previous experiments [27,34,35]. Briefly, HUVEC and astrocytes (50,000 cells/well) positioned in 24-Transwells plates in complete medium were stimulated with plasma for 3 h, as described for the MTT assay.…”

“…The MitoROS release was examined using the Cayman's Mitochondrial ROS Detection Assay Kit (Cayman Chemical) [27,35,36]. In particular, 50,000 HUVEC/astrocytes/well were positioned in 24-Transwells plates in complete medium and they were stimulated with plasma, as was the case for cell viability and mitochondrial membrane potential quantification.…”