title: Transient non-viral cutaneous gene delivery in burn wounds.
abstract: BACKGROUND
  Gene transfer to burn wounds could present an alternative to conventional and often insufficient topical and systemic application of therapeutic agents to aid in wound healing. The goals of this study were to assess and optimize the potential of transient non-viral gene delivery to burn wounds.
  
  
  METHODS
  HaCaT cells were transfected with luciferase or beta-galactosidase transgene using either pure plasmid DNA (pDNA) or complexed with Lipofectamine 2000, FuGENE6, or DOTAP-Chol. Expression was determined by bioluminescence and fluorescence. Forty male Sprague-Dawley rats received naked pDNA, lipoplexes, or carrier control intradermally into either unburned skin, superficial, partial, or full-thickness scald burn. Animals were sacrificed after 24 h, 48 h, or 7 days, and transgene expression was assessed.
  
  
  RESULTS
  Gene transfer to HaCaT cells showed the overall highest expression for DOTAP/Chol (77.85 ng luciferase/mg protein), followed by Lipofectamine 2000 (33.14 ng luciferase/mg protein). pDNA-derived gene transfer to superficial burn wounds showed the highest expression among burn groups (0.77 ng luciferase/mg protein). However, lipoplex-derived gene transfer to superficial burns and unburned skin failed to show higher expression.
  
  
  CONCLUSIONS
  Lipofectamine 2000 and DOTAP/Chol lipoplex showed significantly enhanced gene transfer, whereas no transfection was detectable for naked DNA in vitro. In contrast to the in vitro study, naked DNA was the only agent with which gene delivery was successful in experimental burn wounds. These findings highlight the limited predictability of in vitro analysis for gene delivery as a therapeutic approach.
authors.affiliation: Department for Plastic Surgery, Burn Center, BG University Hospital Bergmannsheil, Ruhr University Bochum, Germany. lars.steinstraesser@ruhr-uni-bochum.de
authors.forename: L
authors.initials: L
authors.lastname: Steinstraesser
authors.forename: T
authors.initials: T
authors.lastname: Hirsch
authors.forename: J
authors.initials: J
authors.lastname: Beller
authors.forename: D
authors.initials: D
authors.lastname: Mittler
authors.forename: M
authors.initials: M
authors.lastname: Sorkin
authors.forename: G
authors.initials: G
authors.lastname: Pazdierny
authors.forename: F
authors.initials: F
authors.lastname: Jacobsen
authors.forename: E
authors.initials: E
authors.lastname: Eriksson
authors.forename: H U
authors.initials: HU
authors.lastname: Steinau
mesh_terms: D000818:Animals; D002056:Burns / Q000628:therapy*; D002460:Cell Line; D018014:Gene Transfer Techniques / Q000592:standards*; D017930:Genes, Reporter; D006801:Humans; D008297:Male; D010714:Phosphatidylethanolamines; D051381:Rats; D017207:Rats, Sprague-Dawley; D012867:Skin / Q000378:metabolism*; D014162:Transfection / Q000379:methods* / Q000592:standards
chemical_list: D010714:Phosphatidylethanolamines; C020888:1,2-dielaidoylphosphatidylethanolamine