Differential effects of bevacizumab, ranibizumab and aflibercept on cell viability, phagocytosis and mitochondrial bioenergetics of retinal pigment epithelial cell

Abstract

ABSTRACT.Purpose: To evaluate the short-and long-term effects of most clinically used anti-vascular endothelial growth factor agents, including bevacizumab, ranibizumab or aflibercept, on cell viability, phagocytosis, mitochondrial bioenergetics and the oxidant acrolein-induced oxidative stress of human adult retinal pigment epithelial (ARPE)-19 cells. Methods: In cultured ARPE-19 cells, cell viability was measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, phagocytotic activity and intracellular reactive oxygen species (ROS) level were determined by flow cytometry, mitochondrial bioenergetics was assessed using a Seahorse XF24 Extracellular Flux Analyzer, and protein expression was measured by Western blotting. Results: Long-term exposure to all three agents had no effect on cell viability; but rescued the ARPE-19 cells from acrolein-induced decrease in cell viability. Bevacizumab, but not ranibizumab or aflibercept, suppressed the phagocytotic activity of ARPE-19 cells and exerted significantly less protection against acroleininduced inhibition of phagocytosis. Both ranibizumab and aflibercept increased basal respiratory rate and maximal mitochondrial respiratory capacity after 1-hr exposure; but returned to baseline following 24-or 72-hr exposure. In contrast, both responses were reduced on short-term exposure, but augmented after long-term exposure to bevacizumab. Long-term pretreatment with all three agents reversed acrolein-induced impairment of mitochondrial bioenergetics, overproduction of ROS and phosphorylation of the mitogen-activated protein kinases in ARPE-19 cells. Conclusion: Bevacizumab might affect mitochondrial bioenergetics differently from that by ranibizumab and aflibercept. Ranibizumab and aflibercept at their therapeutic dose protect against acrolein-induced oxidative cytotoxicity in human ARPE-19 cells via an increase in mitochondrial bioenergetics. An early protective action on mitochondrial bioenergetic capacity might be used to predict possible long-term antioxidative effects of the agents in the eye.

Effects of VEGF inhibitors on human retinal pigment epithelium under high glucose and hypoxia

“…There was no apparent difference on RPE viability for different concentrations of glucose in the presence of hypoxia. Anti‐VEGF drugs have been shown to have a non‐significant effect on RPE and ARPE‐19 viability, and these investigations were not repeated in this study . Glucose appears to have a beneficial effect on the RPE in vitro.…”

“…Anti-VEGF drugs have been shown to have a nonsignificant effect on RPE and ARPE-19 viability, and these investigations were not repeated in this study. [20][21][22][23][24] Glucose appears to have a beneficial effect on the RPE in vitro. High glucose has been associated with increased proliferation of ARPE-19 as well as affecting the morphology of these cells.…”

Anti-Vascular Endothelial Growth Factors Protect Retinal Pigment Epithelium Cells Against Oxidation by Modulating Nitric Oxide Release and Autophagy

“…Hence, in a previous study by Malik et al, [24] neither Ranibizumab or Aflibercept produced any evidence of mitochondrial beneficial effect at clinical doses, whereas mitochondrial toxicity was observed at clinically relevant doses. In contrast, Sheu et al, [32] hypothesized that the early protective action on mitochondrial bioenergetic capacity could predict possible long-term antioxidative effects of Aflibercept and Ranibizumab in the eye.…”

Differential Effects of Bevacizumab, Ranibizumab, and Aflibercept on the Viability and Wound Healing of Corneal Epithelial Cells

“…18 In this study, bevacizumab treatment, but not aflibercept or ranibizumab treatment, suppressed the phagocytic activity of human ARPE-19 cells. Based on the results of cell viability and phagocytic activity assays, it was suggested that the mechanism of action of bevacizumab differs from that of aflibercept and ranibizumab in ARPE-19 cells.…”

“…ranibizumab (0.125, 0.25, or 0.5 mg/mL), or aflibercept (0.125, 0.5, or 2.0 mg/mL). 18 We used 0, 0.25, 0.5, 1.0, and 2.0 mg/mL concentrations, which include the clinical dose for the vitreous. However, we tested HCECs which are different from the previous cells tested ( .…”