Article Text
Abstract
Background: Bevacizumab is an antiangiogenic compound developed to target tumour vessels. Its off-label use in ophthalmology requires in vitro testing on ocular cells.
Aim: To quantify the antipermeability and antiproliferative effects of bevacizumab on cultured choroidal endothelial cells (CECs). It was examined whether deep-freezing of bevacizumab attenuates its antiangiogenic activity.
Methods: Porcine CECs were cultured in permeable insert systems. Permeability of the cell monolayers was quantified by a fluorescent isothiocyanate-dextran assay after treatment with vascular endothelial growth factor (VEGF; 20–100 ng/ml) alone and in combination with bevacizumab (0.1–1 mg/ml). Proliferation of the CECs was tested using a “wound scratch” assay. The experiments were repeated with bevacizumab after freezing at −20°C for 5 days.
Results: Bevacizumab significantly reduced VEGF-induced permeability in a dose-dependant manner. A molar ratio of 2.6:1 of bevacizumab to VEGF was required for complete blocking of VEGF-induced rise in permeability. CEC proliferation was significantly blocked by bevacizumab (0.5 mg/ml). Thawed bevacizumab after deep freezing showed a moderate, but not statistically significant loss in activity.
Conclusion: Bevacizumab significantly reduces VEGF-induced permeability and proliferation of CECs. Freezing and thawing of bevacizumab will affect its biological activity.
- ARMD, age-related macular degeneration
- CEC, choroidal endothelial cell
- CFA, cell-free area
- FDA, Food and Drug Administration
- FITC, fluorescent isothiocyanate
- VEGF, vascular endothelial growth factor
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Footnotes
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Published Online First 19 December 2006
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Competing interests: None declared.
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