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Tatsuo Nagata, Naoya Miyamoto, Hiroyuki Kondo; Safety of Anti-VEGF Drugs on Cultured Corneal Endothelial Cells. Invest. Ophthalmol. Vis. Sci. 2014;55(13):597.
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It has not been determined whether intravitreal anti-vascular endothelial growth factor (VEGF) drugs are toxic for corneal endothelial cells. We examine the toxicity of aflibercept, ranibizumab, pegaptanib, and bevacizumab on cultured corneal endothelial cells.
The cell line of corneal endothelial cells (CECs) was purchased from the American Type Culture Collection as ATCC CRL 2048. The CECs were exposed to 0.5 to 12.0 mg/mL of aflibercept, 0.125 to 3.0 mg/mL of ranibizumab, 0.075 to 1.8 mg/mL of pegaptanib, and 0.25 to 6.0 mg/mL of bevacizumab. Cell viability was evaluated by the WST-8 assay which is based on the activities of dehydrogenases in the cells. Measurements were made 1 and 4 days after exposure. For cytotoxicity testing, confluent cells were cultured in serum-depleted medium, and the WST-8 assay was performed after 24 hours of incubation. Cell morphology was assessed with a phase-contrast microscope after 7 days of exposure with different concentrations of each anti-VEGF drug. Cellular apoptosis was assessed with TUNEL staining. In addition, the cytoadherence of these cells was determined by immunostaining with E- and N-cadherin.
There were no significant changes in the absorbance at 450 nm with the WST-8 assay on CECs for each drug. That was no cytotoxic effect of aflibercept, ranibizumab, pegaptanib and bevacizumab on CECs when used at concentrations of 12.0, 3.0, 1.8, 6.0 mg/mL or lower, respectively. The cells exposed to the anti-VEGF drugs showed no signs of cellular damage and no changes of the cytoadherence compared with controls.
Anti-VEGF drugs such as aflibercept, ranibizumab, pegaptanib and bevacizumab have no significant cytotoxic effect on cultured corneal endothelial cells and do not change the cell integrity in vitro until they exceed the conventional intravitreal dosage by 24-fold.
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