title: Synergistic effect of vascular endothelial growth factor and granulocyte colony-stimulating factor double gene therapy in mouse limb ischemia.
abstract: BACKGROUND
  Vascular endothelial growth factor (VEGF) has mostly been tested to treat ischemic diseases, although the outcomes obtained are not satisfactory. Our hypothesis is that the local transient expression of VEGF and stem cell mobilizer granulocyte colony-stimulating factor (G-CSF) genes in ischemic limbs can complement their activities and be more efficient for limb recovery.
  
  
  METHODS
  Limb ischemia was surgically induced in mice and 50 microg of VEGF and/or G-CSF genes were locally transferred by electroporation. After 3-4 weeks, evidence of necrosis by visual inspection, capillary density, muscle mass, muscle force and hematopoietic cell mobilization were evaluated.
  
  
  RESULTS
  After 4 weeks, 70% and 90% of the animals of the ischemic group (IG) and VEGF-treated group (VG), respectively, presented limb necrosis, in contrast to only 10% observed in the group of mice treated with both VEGF and G-CSF genes (VGG). Recovery of muscle mass and muscle force was higher than 60% in the VGG compared to the non-ischemic group. The mobilization of Sca1+ cells and neutrophils was also higher in the VGG, which may explain the lower level of necrosis observed in this group (22%, in contrast to 70% in the IG). Capillary density and degree of fibrosis were determined in weeks 3 and 4, and also showed a clear benefit as a result of the use of the G-CSF and VEGF genes together.
  
  
  CONCLUSIONS
  Gene therapy using VEGF and G-CSF demonstrated a synergistic effect promoting vessel and tissue repair in mouse hind limb ischemia.
authors.affiliation: Interdisciplinary center for gene therapy, Federal University of São Paulo, São Paulo-SP, 04044-010, Brazil.
authors.forename: Chester Bittencourt
authors.initials: CB
authors.lastname: Sacramento
authors.forename: Flavia Helena
authors.initials: FH
authors.lastname: da Silva
authors.forename: Nance Beyer
authors.initials: NB
authors.lastname: Nardi
authors.forename: Eduardo Gallatti
authors.initials: EG
authors.lastname: Yasumura
authors.forename: José Carlos Costa
authors.initials: JC
authors.lastname: Baptista-Silva
authors.forename: Abram
authors.initials: A
authors.lastname: Beutel
authors.forename: Ruy Ribeiro
authors.initials: RR
authors.lastname: de Campos
authors.forename: Jane Zveiter
authors.initials: JZ
authors.lastname: de Moraes
authors.forename: Hamilton Silva
authors.initials: HS
authors.lastname: Junior
authors.forename: Vivian Yochiko
authors.initials: VY
authors.lastname: Samoto
authors.forename: Radovan
authors.initials: R
authors.lastname: Borojevic
authors.forename: Sang Won
authors.initials: SW
authors.lastname: Han
mesh_terms: D000818:Animals; D005121:Extremities / Q000098:blood supply*; D015316:Genetic Therapy / Q000379:methods*; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics*; D007511:Ischemia / Q000097:blood / Q000209:etiology / Q000628:therapy*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D018482:Muscle, Skeletal / Q000098:blood supply / Q000502:physiology; D018919:Neovascularization, Physiologic / Q000235:genetics; D016491:Peripheral Vascular Diseases / Q000150:complications / Q000628:therapy*; D012038:Regeneration / Q000235:genetics; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics*
chemical_list: D042461:Vascular Endothelial Growth Factor A; D016179:Granulocyte Colony-Stimulating Factor