title: In vivo gene transfer to skin and wound by microseeding.
abstract: BACKGROUND
  Gene transfer to skin has many potential applications but lacks a safe, practical delivery method. This report presents a new technique, microseeding, for in vivo gene transfer to skin and wounds and for DNA-mediated vaccination. The plasmid DNA solution was delivered directly to the target cells of the skin by a set of oscillating solid microneedles driven by a modified tattooing device.
  
  
  METHODS
  Skin and partial-thickness excisional wounds in pigs were microseeded with either hEGF expression plasmid or beta-galactosidase expression plasmid. Human EGF was also delivered by single injection or particle bombardment. hEGF expression in wound fluid and in target tissue was determined by ELISA with anti-hEGF-specific antibodies. Additionally, weanling pigs were microseeded with a hemagglutinin of swine influenza virus expression plasmid and production of anti-HA-specific antibodies was determined by blocking ELISA.
  
  
  RESULTS
  hEGF expression in microseeded partial thickness wounds (5664 pg/site) and skin sites (969 pg/site) peaked 2 days after transfection being four- to seven-fold higher than gene transfer by a single intradermal injection and two- to three-fold higher than particle-mediated gene transfer. The beta-galactosidase-expressing cells were detected in dermis and epidermis. Pigs microseeded with HA expression plasmid were protected from infection by the Swine influenza virus.
  
  
  CONCLUSIONS
  These results demonstrate that microseeding is a simple and effective method for in vivo gene transfer to skin and wounds and is more efficient than single injection and particle-mediated gene transfer.
authors.affiliation: Division of Plastic Surgery, Brigham and Women's Hospital, Boston, Massachusetts, 02115, USA.
authors.forename: E
authors.initials: E
authors.lastname: Eriksson
authors.forename: F
authors.initials: F
authors.lastname: Yao
authors.forename: T
authors.initials: T
authors.lastname: Svensjö
authors.forename: T
authors.initials: T
authors.lastname: Winkler
authors.forename: J
authors.initials: J
authors.lastname: Slama
authors.forename: M D
authors.initials: MD
authors.lastname: Macklin
authors.forename: C
authors.initials: C
authors.lastname: Andree
authors.forename: M
authors.initials: M
authors.lastname: McGregor
authors.forename: V
authors.initials: V
authors.lastname: Hinshaw
authors.forename: W F
authors.initials: WF
authors.lastname: Swain
mesh_terms: D000818:Animals; D004815:Epidermal Growth Factor / Q000235:genetics; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D018014:Gene Transfer Techniques; D006649:Histocompatibility Antigens / Q000235:genetics; D009980:Influenza A virus; D009976:Orthomyxoviridae Infections; D010957:Plasmids; D012879:Skin Physiological Phenomena; D013552:Swine; D013653:Tattooing / Q000379:methods; D014611:Vaccination / Q000379:methods; D014945:Wound Healing / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D006649:Histocompatibility Antigens; D004815:Epidermal Growth Factor; D001616:beta-Galactosidase