In vivo gene transfer to skin and wound by microseeding

J Surg Res. 1998 Aug;78(2):85-91. doi: 10.1006/jsre.1998.5325.

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.

Materials and 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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Epidermal Growth Factor / genetics
  • Female
  • Gene Expression Regulation, Enzymologic
  • Gene Transfer Techniques*
  • Histocompatibility Antigens / genetics
  • Influenza A virus*
  • Orthomyxoviridae Infections*
  • Plasmids*
  • Skin Physiological Phenomena
  • Swine
  • Tattooing / methods
  • Vaccination / methods
  • Wound Healing / genetics*
  • beta-Galactosidase / genetics

Substances

  • Histocompatibility Antigens
  • Epidermal Growth Factor
  • beta-Galactosidase