Background: Bevacizumab is an anti-angiogenic compound developed to target tumor vessels. Off-label use in ophthalmology requires in vitro testing on ocular cells. Consequently we quantified the anti-permeability and anti-proliferative effect of bevacizumab on cultured choroidal endothelial cells. It was examined whether deep-freezing of bevacizumab attenuates its anti- angiogenic activity.
Methods: Porcine choroidal endothelial cells (CEC) were cultured in permeable insert systems. Permeability of the cell monolayers was quantified in an FITC-dextran assay after treatment with VEGF (20-100 ng/ml) alone and in combination with bevacizumab (0.1 & [minus]1 mg/ml). Proliferation of the CEC was tested using a wound scratch assay. The experiments were repeated with bevacizumab after -20C freezing 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 statistically not significant loss in activity.
Conclusion: Bevacizumab significantly reduces VEGF-induced permeability and proliferation of choroidal endothelial cells. Freezing and thawing of bevacizumab will affect its biological activity.
- choroidal endothelial cells