title: Transglutaminase binding fusion protein linked to SLPI reduced corneal inflammation and neovascularization.
abstract: BACKGROUND
  To study the effect of topical administration of a fusion protein (PF-MC) made up of N-terminal portion of the protease inhibitor Trappin-2 (which is a substrate of transglutaminasa-2) and SLPI (protein with anti-inflammatory, anti-bacterial and anti-viral ability), in an animal model of corneal inflammation and angiogenesis.
  
  
  METHODS
  An alkali injury was produced with a filter paper of 3 mm with 1 N NaOH during 40 seconds on the right cornea of 36 male Sprague Dawley rats, under general anesthesia. Animals were divided into three groups according to treatment. Group 1 was treated with 10 ul of PF-MC (200 ug/ml; n = 12), Group 2, with 10 ul of SLPI (200 ug/ml; n = 12) and Group 3 was treated with buffer (10 ul; n = 12) topically administered four times a day for up to 7 days. Half of the animals were sacrificed at day 3 before making a re-epithelialization time analysis with fluorescein staining at 18 and 24 hours. In the remaining animals corneal opacity was studied and digital photographs were taken at day 7 before doing euthanasia. Eyes were processed for histology and immunofluorescence.
  
  
  RESULTS
  Corneal ulcerated area was significantly lower in PF-MC treated animals compared to SLPI and buffer-treated animals at 18 hours and 24 hours postinjury. A clear cornea and fundus red reflex was only found among PF-MC treated animals. Histological analysis revealed a stratified corneal epithelium with at least three layers in all PF-MC animals at day 7. In this group there was a reduced number of PMNs in the corneal stroma at 3 and 7 days of follow-up. Besides, corneal neovascularization was much more extended in SLPI and Buffer animals than in animals treated with PF-MC.
  
  
  CONCLUSIONS
  The binding of SLPI with Cementoin to transglutaminase seems to be an effective strategy to treat corneal inflammation and angiogenesis.
authors: Nanomedicine & Vision Group, Faculty of Biomedical Sciences, Austral University, Juan Domingo Perón, 1500 1629 Pilar, Buenos Aires, Argentina.|Department of Ophthalmology, Austral University Hospital, Pilar, Buenos Aires, Argentina.
authors: Juan P
authors: JP
authors: Salica
authors: Laboratory of Immunomodulators, School of Medicine, Cefybo (Conicet-University of Buenos Aires), Buenos Aires, Argentina.
authors: Diego
authors: D
authors: Guerrieri
authors: Laboratory of Immunomodulators, School of Medicine, Cefybo (Conicet-University of Buenos Aires), Buenos Aires, Argentina.
authors: Paulo
authors: P
authors: Maffia
authors: Department of Ocular Pathology, Fundación Oftalmológica Argentina "Jorge Malbrán", Buenos Aires, Argentina.
authors: Juan O
authors: JO
authors: Croxatto
authors: Laboratory of Immunomodulators, School of Medicine, Cefybo (Conicet-University of Buenos Aires), Buenos Aires, Argentina.
authors: H Eduardo
authors: HE
authors: Chuluyan
authors: Nanomedicine & Vision Group, Faculty of Biomedical Sciences, Austral University, Juan Domingo Perón, 1500 1629 Pilar, Buenos Aires, Argentina. jgallo06@gmail.com.|Department of Ophthalmology, Austral University Hospital, Pilar, Buenos Aires, Argentina. jgallo06@gmail.com.
authors: Juan E
authors: JE
authors: Gallo
mesh_terms: D000287:Administration, Topical; D000818:Animals; D002057:Burns, Chemical / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D002452:Cell Count; D016510:Corneal Neovascularization / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D004195:Disease Models, Animal; D019573:Epithelium, Corneal / Q000502:physiology; D005126:Eye Burns / Q000139:chemically induced*; D019084:Fluorescent Antibody Technique, Indirect; D019204:GTP-Binding Proteins / Q000235:genetics*; D007634:Keratitis / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D008297:Male; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D051381:Rats; D017207:Rats, Sprague-Dawley; D062326:Re-Epithelialization; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage*; D053481:Secretory Leukocyte Peptidase Inhibitor / Q000235:genetics*; D011503:Transglutaminases / Q000235:genetics*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; C506814:SLPI protein, human; D053481:Secretory Leukocyte Peptidase Inhibitor; D042461:Vascular Endothelial Growth Factor A; C467498:vascular endothelial growth factor A, rat; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D019204:GTP-Binding Proteins
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