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

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• 作者：Manuel Saenz-de-Viteri^a ; ^b ; ¹ ; Patricia Ferná ; ndez-Robredo^a ; ^c ; ¹ ; Marí ; a Herná ; ndez^a ; ^c ; ^{mahersan@unav.es" class="auth_mail" title="E-mail the corresponding author} ; Jaione Bezunartea^a ; Nicholas Reiter^a ; Sergio Recalde^a ; ^c ; ² ; Alfredo Garcí ; a-Layana^a ; ^b ; ^c ; ²
• 关键词：Vascular endothelial growth factor ; Retinal pigmented epithelium ; VEGF-antagonists ; Age-related-macular degeneration ; Diabetic macular edema
• 刊名：Biochemical Pharmacology
• 出版年：2016
• 出版时间：1 March 2016
• 年：2016
• 卷：103
• 期：Complete
• 页码：129-139
• 全文大小：2138 K

文摘

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.