title: Effect of recombinant adeno-associated virus vector-mediated vascular endothelial growth factor gene transfer on wound healing after burn injury.
abstract: OBJECTIVE
  The purpose of this study was to investigate the effect of recombinant adeno-associated viral (rAAV) vector-mediated human vascular endothelial growth factor (VEGF165) transfer on experimental burn wounds.
  
  
  METHODS
  Randomized experiment.
  
  
  METHODS
  Research laboratory.
  
  
  METHODS
  C57BL/6 male mice weighing 25-30 g.
  
  
  METHODS
  Mice were immersed in 80 degrees C water for 10 secs to achieve a partial-thickness scald burn. Animals were randomized to receive at two injection sites on the edge of the burn either 1011 copies of the rAAV-VEGF165 or the vector carrying the control and inert gene beta-galactosidase (rAAV-LacZ). On day 14 the animals were killed. Burn areas were used for histologic examination, evaluation of VEGF expression (immunohistochemistry) and VEGF wound content (enzyme-linked immunosorbent assay), determination of wound nitrite, and measurement of messenger RNA (mRNA) for endothelial and inducible nitric oxide synthase (eNOS and iNOS).
  
  
  RESULTS
  rAAV-VEGF165 increased epithelial proliferation, angiogenesis, and maturation of the extracellular matrix. Furthermore, gene transfer enhanced VEGF expression, studied by immunohistochemistry, and the wound content of the mature protein (rAAV-LacZ, 11 +/- 5 pg/wound; rAAV-VEGF165, 104 +/- 7 pg/wound). Moreover, VEGF165 gene transfer increased wound content of nitrate. Finally, rAAV-VEGF165 administration enhanced the messenger RNA for eNOS (rAAV-VEGF165, 1.1 +/- 0.2 relative amount of eNOS mRNA; rAAV-LacZ, 0.66 +/- 0.3 relative amount of eNOS mRNA) and iNOS (rAAV-VEGF165, 0.8 +/- 0.09 relative amount of iNOS mRNA; rAAV-LacZ, 0.45 +/- 0.05 relative amount of iNOS mRNA).
  
  
  CONCLUSIONS
  Our study suggests that rAAV-VEGF gene transfer may be an effective therapeutic approach to improve clinical outcomes after thermal injury.
authors.affiliation: Department of Surgical Sciences, Section of Plastic Surgery, University of Messina, Italy.
authors.forename: Mariarosaria
authors.initials: M
authors.lastname: Galeano
authors.forename: Barbara
authors.initials: B
authors.lastname: Deodato
authors.forename: Domenica
authors.initials: D
authors.lastname: Altavilla
authors.forename: Giovanni
authors.initials: G
authors.lastname: Squadrito
authors.forename: Paolo
authors.initials: P
authors.lastname: Seminara
authors.forename: Herbert
authors.initials: H
authors.lastname: Marini
authors.forename: Francesco
authors.initials: F
authors.lastname: Stagno d'Alcontres
authors.forename: Michele
authors.initials: M
authors.lastname: Colonna
authors.forename: Margherita
authors.initials: M
authors.lastname: Calò
authors.forename: Patrizia
authors.initials: P
authors.lastname: Lo Cascio
authors.forename: Valerio
authors.initials: V
authors.lastname: Torre
authors.forename: Mauro
authors.initials: M
authors.lastname: Giacca
authors.forename: Francesco S
authors.initials: FS
authors.lastname: Venuti
authors.forename: Francesco
authors.initials: F
authors.lastname: Squadrito
mesh_terms: D000818:Animals; D002056:Burns / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology / Q000628:therapy*; D000229:Dependovirus; D016228:Endothelial Growth Factors / Q000235:genetics* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D007150:Immunohistochemistry; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics* / Q000378:metabolism; D008222:Lymphokines / Q000235:genetics* / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D019001:Nitric Oxide Synthase / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; D012867:Skin / Q000378:metabolism / Q000473:pathology; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D014945:Wound Healing / Q000502:physiology*; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D016228:Endothelial Growth Factors; D036341:Intercellular Signaling Peptides and Proteins; D008222:Lymphokines; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; C496317:Nos2 protein, mouse; C496315:Nos3 protein, mouse; D001616:beta-Galactosidase