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Anti-permeability and anti-proliferative effects of standard and frozen bevacizumab on choroidal endothelial cells
  1. Swaantje Peters (swaantje.peters{at}gmx.de),
  2. Sylvie Julien (sylvie.julien{at}med.uni-tuebingen.de),
  3. Peter Heiduschka (peter.heiduschka{at}med.uni-tuebingen.de),
  4. Salvatore Grisanti (salvatore.grisanti{at}med.uni-tuebingen.de),
  5. Focke Ziemssen (focke.ziemssen{at}med.uni-tuebingen.de),
  6. Martin Adler (martin.adler{at}med.uni-tuebingen.de),
  7. Ulrich Schraermeyer (ulrich.schraermeyer{at}med.uni-tuebingen.de),
  8. Karl U. Bartz-Schmidt (u.bartz-schmidt{at}med.uni-tuebingen.de),
  9. The Tuebingen Bevacizumab Study Group
  1. University Eye Hospital Tuebingen, Germany
  2. University Eye Hospital Tuebingen, Germany
  3. University Eye Hospital Tuebingen, Germany
  4. University Eye Hospital Tuebingen, Germany
  5. University Eye Hospital Tuebingen, Germany
  6. University Eye Hospital Tuebingen, Germany
  7. University Eye Hospital Tuebingen, Germany
  8. University Eye Hospital Tuebingen, Germany
  9. University Eye Hospital Tuebingen, Germany

    Abstract

    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.

    • VEGF
    • bevacizumab
    • choroidal endothelial cells
    • permeability
    • proliferation

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