title: Antipermeability and antiproliferative effects of standard and frozen bevacizumab on choroidal endothelial cells.
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.
  
  
  OBJECTIVE
  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 degrees 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.
  
  
  CONCLUSIONS
  Bevacizumab significantly reduces VEGF-induced permeability and proliferation of CECs. Freezing and thawing of bevacizumab will affect its biological activity.
authors.affiliation: University Eye Hospital, Eberhard-Karls University Tuebingen, Schleichstrasse 12, D-72076 Tuebingen, Germany. swaantje.peters@gmx.de
authors.forename: Swaantje
authors.initials: S
authors.lastname: Peters
authors.forename: Sylvie
authors.initials: S
authors.lastname: Julien
authors.forename: Peter
authors.initials: P
authors.lastname: Heiduschka
authors.forename: Salvatore
authors.initials: S
authors.lastname: Grisanti
authors.forename: Focke
authors.initials: F
authors.lastname: Ziemssen
authors.forename: Martin
authors.initials: M
authors.lastname: Adler
authors.forename: Ulrich
authors.initials: U
authors.lastname: Schraermeyer
authors.forename: Karl-Ulrich
authors.initials: KU
authors.lastname: Bartz-Schmidt
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D061067:Antibodies, Monoclonal, Humanized; D000068258:Bevacizumab; D002199:Capillary Permeability / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D002829:Choroid / Q000098:blood supply / Q000166:cytology / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004355:Drug Stability; D042783:Endothelial Cells / Q000187:drug effects; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects*; D005615:Freezing; D013552:Swine
chemical_list: D020533:Angiogenesis Inhibitors; D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D000068258:Bevacizumab