Electroporation enhances transfection efficiency in murine cutaneous wounds.
Transfection of wounds with DNA-encoding growth factors has the potential to improve healing, but current means of nonviral gene delivery are inefficient. Repeated high doses of DNA, necessary to achieve reliable gene expression, are detrimental to healing. We assessed the ability of in vivo electroporation to enhance gene expression. Full-thickness cutaneous excisional wounds were created on the dorsum of female mice. A luciferase- encoding plasmid driven by a CMV promoter was injected at the wound border. Following plasmid administration, electroporative pulses were applied to injection sites. Pulse parameters were varied over a range of voltage, duration, and number. Animals were euthanized at intervals after transfection and the luciferase activity measured. Application of electric pulses consistently increased luciferase expression. The electroporative effect was most marked at a plasmid dose of 50 micro g, where an approximate tenfold increase was seen. Six 100- micro s-duration pulses of 1750 V/cm were found to be the most effective in increasing luciferase activity. High numbers of pulses tended to be less effective than smaller numbers. This optimal electroporation regimen had no detrimental effect on wound healing. We conclude that electroporation increases the efficiency of transgene expression and may have a role in gene therapy to enhance wound healing.
Byrnes
Colman K
CK
Section of Surgical Sciences, Johns Hopkins Medical Institutions, Johns Hopkins Bayview Medical Center, 4940 Eastern Avenue, Baltimore, MD 21224, USA.
Malone
Robert W
RW
Akhter
Nabeel
N
Nass
Petra H
PH
Wetterwald
Antoinette
A
Cecchini
Marco G
MG
Duncan
Mark D
MD
Harmon
John W
JW
D000818:Animals; D018274:Electroporation; D005260:Female; D015870:Gene Expression; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids; D012867:Skin / Q000293:injuries; D014162:Transfection / Q000379:methods*; D014945:Wound Healing / Q000235:genetics / Q000502:physiology*
D008156:Luciferases