title: Topical Ranibizumab inhibits inflammatory corneal hem- and lymphangiogenesis.
abstract: OBJECTIVE
  Ranibizumab (Lucentis(®) ) is a Fab-Fragment of a recombinant, humanized, monoclonal VEGF (anti-vascular endothelial growth factor) antibody. This study analyzed the ability of topical Ranibizumab to inhibit lymphangiogenesis in addition to hemangiogenesis after acute corneal inflammation in vivo. In addition, the effect of Ranibizumab on the proliferation of human lymphatic endothelial cells (LECs) and blood endothelial cells (BECs) in vitro was studied.
  
  
  METHODS
  The inhibitory effect of Ranibizumab on LECs and BECs was studied in vitro using a proliferation enzyme-linked immunosorbent assay (ELISA) assay. To study the in vivo effects of Ranibizumab, the mouse model of suture induced inflammatory corneal neovascularization was used. Study mice received topical Ranibizumab as eye drops. After 1 week excised corneas were stained with LYVE-1 and CD31. Hemangiogenesis and lymphangiogenesis were analyzed morphometrically by using a semiautomatic method based on the image analyzing program Cell^F.
  
  
  RESULTS
  An antiproliferative effect of Ranibizumab was seen in vitro on both human BECs and LECs with a significance of p < 0.0001 and p < 0.0004, respectively. In vivo experiments showed that topical application of Ranibizumab significantly inhibits both hemangiogenesis (p = 0.0026) and lymphangiogenesis (p = 0.0026) in the cornea.
  
  
  CONCLUSIONS
  Ranibizumab is a potent inhibitor of inflammatory corneal hemangiogenesis and lymphangiogenesis in vivo with a direct inhibitory effect on both endothelial cell types in vitro. This study for the first time demonstrates an inhibitory effect of Ranibizumab on lymphatic vessels which could have a wider range of clinical applications.
authors.affiliation: Department of Ophthalmology, University of Cologne, Cologne, GermanyDepartment of Ophthalmology, University of Erlangen-Nürnberg, Erlangen, GermanyCornea Service, Clinical Research Centre, Vasan Eye Care Hospital, Chennai, IndiaT.N Shukla Eye Hospital, Ranjhi, Jabalpur, India.
authors.forename: Franziska
authors.initials: F
authors.lastname: Bucher
authors.forename: Anand
authors.initials: A
authors.lastname: Parthasarathy
authors.forename: Antonio
authors.initials: A
authors.lastname: Bergua
authors.forename: Jasmine
authors.initials: J
authors.lastname: Onderka
authors.forename: Birgit
authors.initials: B
authors.lastname: Regenfuss
authors.forename: Claus
authors.initials: C
authors.lastname: Cursiefen
authors.forename: Felix
authors.initials: F
authors.lastname: Bock
mesh_terms: D000287:Administration, Topical; D020533:Angiogenesis Inhibitors / Q000008:administration & dosage / Q000494:pharmacology*; D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000008:administration & dosage / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D016510:Corneal Neovascularization / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D004195:Disease Models, Animal; D004729:Endothelium, Lymphatic / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006023:Glycoproteins / Q000378:metabolism; D042583:Lymphangiogenesis / Q000187:drug effects*; D026901:Membrane Transport Proteins; D051379:Mice; D008807:Mice, Inbred BALB C; D009883:Ophthalmic Solutions; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D000069579:Ranibizumab; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors
chemical_list: D020533:Angiogenesis Inhibitors; D061067:Antibodies, Monoclonal, Humanized; D006023:Glycoproteins; D026901:Membrane Transport Proteins; D009883:Ophthalmic Solutions; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D000069579:Ranibizumab; D042461:Vascular Endothelial Growth Factor A; C492273:LYVE1 protein, mouse; C467480:vascular endothelial growth factor A, mouse