Single- and repeated-dose toxicity study of bevacizumab, ranibizumab, and aflibercept in ARPE-19 cells under normal and oxidative stress conditions.

Biochem Pharmacol 2016 Mar 12;103:129-39. Epub 2016 Jan 12.

Experimental Ophthalmology Laboratory, School of Medicine, University of Navarra, 1 Irunlarrea Street, 31008 Pamplona, Spain; Department of Ophthalmology, Clínica Universidad de Navarra, School of Medicine, University of Navarra, 36 Pio XII Avenue, 31008 Pamplona, Spain; IdiSNA, Navarra Institute for Health Research, 31008 Pamplona, Spain.

We assessed the effect of single and repeated doses of bevacizumab, ranibizumab, and aflibercept on cell viability, proliferation, permeability, and apoptosis of ARPE-19 cells. MTT and BrdU assays were used to determine viability and proliferation after single or repeated doses of anti-VEGF drugs under normal and oxidative stress conditions. Caspase-3 expression after single and repeated doses of the 3 drugs was assessed using immunofluorescence. Transepithelial-electrical-resistance (TER) was measured to study the effect of anti-VEGFs on retinal pigment epithelium (RPE) permeability under normal and oxidative stress conditions. Flow cytometry was used to detect intracellular accumulation of the drugs. Finally, a wound healing assay was performed to investigate the effect of the drugs on RPE cell migration. Single and multiple doses of anti-VEGF drugs had no effect on cell viability and proliferation. The oxidative effect of H2O2 decreased cell viability and proliferation; however, no difference was observed between anti-VEGF treatments. Immunofluorescence performed after single and repeated doses of the drugs revealed some caspase-3 expression. Interestingly, anti-VEGFs restored the increased permeability induced by H2O2. The 3 drugs accumulated inside the cells and were detectable 5 days after treatment. Finally, none of the drugs affected migration. In conclusion, no measureable toxic effect was observed after single or repeated doses of VEGF antagonists under normal and oxidative stress. Intracellular accumulation of the drugs does not seem to be toxic or affect cell functions. Our study suggests that anti-VEGFs could have a preventive effect on the maintenance of the RPE barrier under oxidative stress.

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.bcp.2015.12.017DOI Listing
March 2016

Publication Analysis

Top Keywords

oxidative stress
20
single repeated
20
repeated doses
20
normal oxidative
16
viability proliferation
16
stress conditions
12
cell viability
12
drugs
9
caspase-3 expression
8
doses anti-vegf
8
intracellular accumulation
8
accumulation drugs
8
anti-vegf drugs
8
arpe-19 cells
8
doses drugs
8
ranibizumab aflibercept
8
bevacizumab ranibizumab
8
single
6
doses
6
oxidative
6

Similar Publications

Effects of Bevacizumab, Ranibizumab, and Aflibercept on MicroRNA Expression in a Retinal Pigment Epithelium Cell Culture Model of Oxidative Stress.

J Ocul Pharmacol Ther 2018 05 1;34(4):346-353. Epub 2018 Feb 1.

3 Department of Pharmacology, Faculty of Medicine, Kahramanmaraş Sütçü İmam University , Kahramanmaraş, Turkey .

Purpose: This study aimed to evaluate the effects of bevacizumab, ranibizumab, and aflibercept on the microRNA (miRNA) expression in human retinal pigment epithelium cell (ARPE-19) culture model of oxidative stress.

Methods: Control cells were cultured in the hydrogen peroxide (HO)-free medium. In HO group ARPE-19 cells were exposed to 600 μM HO alone for 18 h. Read More

View Article and Full-Text PDF
May 2018

Comparative toxicity and proliferation testing of aflibercept, bevacizumab and ranibizumab on different ocular cells.

Br J Ophthalmol 2013 Jul 17;97(7):917-23. Epub 2013 May 17.

Centre of Ophthalmology, University Eye Hospital Tübingen, Tübingen, Germany.

Background/aims: Vascular endothelial growth factor (VEGF) is a key factor in the pathogenesis of neovascular retinal diseases including age-related macular degeneration. VEGF inhibitors including ranibizumab, pegaptanib or bevacizumab improve retinal morphology and vision in many patients. The recently approved drug aflibercept (VEGF Trap-Eye/Eyelea, Regeneron, Tarrytown, New York, USA) offers a new therapy modality. Read More

View Article and Full-Text PDF
July 2013

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

Acta Ophthalmol 2015 Dec 14;93(8):e631-43. Epub 2015 May 14.

Center for Translational Research in Biomedical Sciences, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan.

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. Read More

View Article and Full-Text PDF
December 2015

Fc Receptor Inhibition Reduces Susceptibility to Oxidative Stress in Human RPE Cells Treated with Bevacizumab, but not Aflibercept.

Cell Physiol Biochem 2016 15;38(2):737-47. Epub 2016 Feb 15.

Department of Ophthalmology, University of Lx00FC;beck, Lx00FC;beck, Germany.

Background/aims: VEGF-A is induced by oxidative stress, and functions as a survival factor for various cell types, including retinal pigment epithelial (RPE) cells. Anti-vascular endothelial growth factor (VEGF) drugs like aflibercept and bevacizumab have shown to be most effective in treating neovascular age-related macular degeneration (AMD), however uptake of the drugs might lead to interference with cell physiology. Herein, we evaluated the significance of the Fc receptor (FcR) within this context and moreover explored the impact of VEGF inhibition under normal conditions as well as under oxidative stress, in terms of potential adverse effects. Read More

View Article and Full-Text PDF
November 2016