title: Effects of aflibercept on primary RPE cells: toxicity, wound healing, uptake and phagocytosis.
abstract: OBJECTIVE
  Anti-VEGF treatment is the therapy of choice in age-related macular degeneration, and is also applied in diabetic macular oedema or retinal vein occlusion. Recently, the fusion protein, aflibercept, has been approved for therapeutic use. In this study, we investigate the effects of aflibercept on primary RPE cells.
  
  
  METHODS
  Primary RPE cells were prepared from freshly slaughtered pigs' eyes. The impact of aflibercept on cell viability was investigated with MTT and trypan blue exclusion assay. The influence of aflibercept on wound healing was assessed with a scratch assay. Intracellular uptake of aflibercept was investigated in immunohistochemistry and its influence on phagocytosis with a phagocytosis assay using opsonised latex beads.
  
  
  RESULTS
  Aflibercept displays no cytotoxicity on RPE cells but impairs its wound healing ability. It is taken up into RPE cells and can be intracellularly detected for at least 7 days. Intracellular aflibercept impairs the phagocytic capacity of RPE cells.
  
  
  CONCLUSIONS
  Aflibercept interferes with the physiology of RPE cells, as it is taken up into RPE cells, which is accompanied by a reduction of the phagocytic ability. Additionally, it impairs the wound healing capacity of RPE cells. These effects on the physiology of RPE cells may indicate possible side effects.
authors.affiliation: Department of Ophthalmology, University of Kiel, University Medical Center, Kiel, Germany.
authors.forename: Alexa
authors.initials: A
authors.lastname: Klettner
authors.affiliation: Department of Ophthalmology, University of Kiel, University Medical Center, Kiel, Germany.
authors.forename: Nihat
authors.initials: N
authors.lastname: Tahmaz
authors.affiliation: Department of Ophthalmology, University of Kiel, University Medical Center, Kiel, Germany.
authors.forename: Michaela
authors.initials: M
authors.lastname: Dithmer
authors.affiliation: Department of Ophthalmology, University of Kiel, University Medical Center, Kiel, Germany.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Richert
authors.affiliation: Department of Ophthalmology, University of Kiel, University Medical Center, Kiel, Germany.
authors.forename: Johann
authors.initials: J
authors.lastname: Roider
mesh_terms: D020533:Angiogenesis Inhibitors / Q000633:toxicity; D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000633:toxicity; D000068258:Bevacizumab; D002465:Cell Movement / Q000187:drug effects; D002470:Cell Survival; D002478:Cells, Cultured; D004396:Coloring Agents / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D008863:Microspheres; D010587:Phagocytosis / Q000187:drug effects*; D000069579:Ranibizumab; D040262:Receptors, Vascular Endothelial Growth Factor / Q000633:toxicity*; D011993:Recombinant Fusion Proteins / Q000633:toxicity*; D055213:Retinal Pigment Epithelium / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D013552:Swine; D014343:Trypan Blue / Q000378:metabolism; D014945:Wound Healing / Q000187:drug effects*
chemical_list: D020533:Angiogenesis Inhibitors; D061067:Antibodies, Monoclonal, Humanized; D004396:Coloring Agents; D011993:Recombinant Fusion Proteins; C533178:aflibercept; D000068258:Bevacizumab; D040262:Receptors, Vascular Endothelial Growth Factor; D014343:Trypan Blue; D000069579:Ranibizumab
keywords: Pharmacology; Retina; Wound healing