title: Adenoviral-mediated gene transfer of insulin-like growth factor 1 enhances wound healing and induces angiogenesis.
abstract: BACKGROUND
  Chronic wounds are characterized by a wound healing and neovascularization deficit. Strategies to increase neovascularization can significantly improve chronic wound healing. Insulin-like growth factor (IGF)-1 is reported to be a keratinocyte mitogen and is believed to induce angiogenesis via a vascular endothelial growth factor (VEGF)-dependent pathway. Using a novel ex vivo human dermal wound model and a diabetic-impaired wound healing murine model, we hypothesized that adenoviral overexpression of IGF-1 (Ad-IGF-1) will enhance wound healing and induce angiogenesis through a VEGF-dependent pathway.
  
  
  METHODS
  Ex vivo: 6-mm full-thickness punch biopsies were obtained from normal human skin, and 3-mm full-thickness wounds were created at the center. Skin explants were maintained at air liquid interface. Db/db murine model: 8-mm full-thickness dorsal wounds in diabetic (db/db) mice were created. Treatment groups in both human ex vivo and in vivo db/db wound models include 1×10(8) particle forming units of Ad-IGF-1 or Ad-LacZ, and phosphate buffered saline (n=4-5/group). Cytotoxicity (lactate dehydrogenase) was quantified at days 3, 5, and 7 for the human ex vivo wound model. Epithelial gap closure (hematoxylin and eosin; Trichrome), VEGF expression (enzyme-linked immunosorbent assay), and capillary density (CD 31+CAPS/HPF) were analyzed at day 7.
  
  
  RESULTS
  In the human ex vivo organ culture, the adenoviral vectors did not demonstrate any significant difference in cytotoxicity compared with phosphate buffered saline. Ad-IGF-1 overexpression significantly increases basal keratinocyte migration, with no significant effect on epithelial gap closure. There was a significant increase in capillary density in the Ad-IGF-1 wounds. However, there was no effect on VEGF levels in Ad-IGF-1 samples compared with controls. In db/db wounds, Ad-IGF-1 overexpression significantly improves epithelial gap closure and granulation tissue with a dense cellular infiltrate compared with controls. Ad-IGF-1 also increases capillary density, again with no significant difference in VEGF levels in the wounds compared with control treatments.
  
  
  CONCLUSIONS
  In two different models, our data demonstrate that adenoviral-mediated gene transfer of IGF-1 results in enhanced wound healing and induces angiogenesis via a VEGF-independent pathway. Understanding the underlying mechanisms of IGF-1 effects on angiogenesis may help produce novel therapeutics for chronic wounds or diseases characterized by a deficit in neovascularization.
authors.affiliation: Division of Pediatric, General, Thoracic and Fetal Surgery, Laboratory for Regenerative Wound Healing, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio.
authors.forename: Swathi
authors.initials: S
authors.lastname: Balaji
authors.affiliation: Division of Pediatric, General, Thoracic and Fetal Surgery, Laboratory for Regenerative Wound Healing, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio.
authors.forename: Maria
authors.initials: M
authors.lastname: LeSaint
authors.affiliation: Division of Pediatric, General, Thoracic and Fetal Surgery, Laboratory for Regenerative Wound Healing, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio.
authors.forename: Sukanta S
authors.initials: SS
authors.lastname: Bhattacharya
authors.affiliation: Division of Pediatric, General, Thoracic and Fetal Surgery, Laboratory for Regenerative Wound Healing, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio.
authors.forename: Chad
authors.initials: C
authors.lastname: Moles
authors.affiliation: Division of Pediatric, General, Thoracic and Fetal Surgery, Laboratory for Regenerative Wound Healing, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio.
authors.forename: Yashu
authors.initials: Y
authors.lastname: Dhamija
authors.affiliation: Division of Pediatric, General, Thoracic and Fetal Surgery, Laboratory for Regenerative Wound Healing, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio.
authors.forename: Mykia
authors.initials: M
authors.lastname: Kidd
authors.affiliation: Division of Pediatric, General, Thoracic and Fetal Surgery, Laboratory for Regenerative Wound Healing, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio.
authors.forename: Louis D
authors.initials: LD
authors.lastname: Le
authors.affiliation: Division of Pediatric, General, Thoracic and Fetal Surgery, Laboratory for Regenerative Wound Healing, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio.
authors.forename: Alice
authors.initials: A
authors.lastname: King
authors.affiliation: Division of Pediatric, General, Thoracic and Fetal Surgery, Laboratory for Regenerative Wound Healing, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio.
authors.forename: Aimen
authors.initials: A
authors.lastname: Shaaban
authors.affiliation: Center for Children's Surgery, Children's Hospital Colorado and the University of Colorado, School of Medicine, Aurora, Colorado.
authors.forename: Timothy M
authors.initials: TM
authors.lastname: Crombleholme
authors.affiliation: Department of Medicine, Stanford University, Palo Alto, California.
authors.forename: Paul
authors.initials: P
authors.lastname: Bollyky
authors.affiliation: Division of Pediatric, General, Thoracic and Fetal Surgery, Laboratory for Regenerative Wound Healing, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio. Electronic address: sundeep.keswani@cchmc.org.
authors.forename: Sundeep G
authors.initials: SG
authors.lastname: Keswani
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000328:Adult; D000818:Animals; D002465:Cell Movement; D005260:Female; D018014:Gene Transfer Techniques; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000235:genetics*; D015603:Keratinocytes / Q000502:physiology; D051379:Mice; D008875:Middle Aged; D018919:Neovascularization, Physiologic; D042461:Vascular Endothelial Growth Factor A / Q000032:analysis; D014945:Wound Healing
chemical_list: D042461:Vascular Endothelial Growth Factor A; D007334:Insulin-Like Growth Factor I
keywords: Diabetes; Gene therapy; IGF-1; Neovascularization; Organ culture; Wound healing; db/db
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