In vivo gene transfer to skin and wound by microseeding.
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.
Eriksson
E
E
Division of Plastic Surgery, Brigham and Women's Hospital, Boston, Massachusetts, 02115, USA.
Yao
F
F
Svensjö
T
T
Winkler
T
T
Slama
J
J
Macklin
M D
MD
Andree
C
C
McGregor
M
M
Hinshaw
V
V
Swain
W F
WF
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
D006649:Histocompatibility Antigens; D004815:Epidermal Growth Factor; D001616:beta-Galactosidase