Delivery of plasmid DNA expression vector for keratinocyte growth factor-1 using electroporation to improve cutaneous wound healing in a septic rat model.
We have previously shown that wound healing was improved in a diabetic mouse model of impaired wound healing following transfection with keratinocyte growth factor-1 (KGF-1) cDNA. We now extend these findings to the characterization of the effects of DNA plasmid vectors delivered to rats using electroporation (EP) in vivo in a sepsis-based model of impaired wound healing. To assess plasmid transfection and wound healing, gWIZ luciferase and PCDNA3.1/KGF-1 expression vectors were used, respectively. Cutaneous wounds were produced using an 8 mm-punch biopsy in Sprague-Dawley rats in which healing was impaired by cecal ligation-induced sepsis. We used National Institutes of Health image analysis software and histologic assessment to analyze wound closure and found that EP increased expression of gWIZ luciferase vector up to 53-fold compared with transfection without EP (p < 0.001). EP-assisted plasmid transfection was found to be localized to skin. Septic rats had a 4.7 times larger average wound area on day 9 compared with control (p < 0.001). Rats that underwent PCDNA3.1/KGF-1 transfection with EP had 60% smaller wounds on day 12 compared with vector without EP (p < 0.009). Quality of healing with KGF-1 vector plus EP scored 3.0 +/- 0.3 and was significantly better than that of 1.8 +/- 0.3 for treatment with vector alone (p < 0.05). We conclude that both the rate and quality of healing were improved with DNA plasmid expression vector for growth factor delivered with EP to septic rats.
Lin
Michael P
MP
Section of Surgical Sciences, Johns Hopkins Bayview Medical Center, Johns Hopkins Medical Institutions, Baltimore, Maryland 21224, USA.
Marti
Guy P
GP
Dieb
Rami
R
Wang
Jiaai
J
Ferguson
Mark
M
Qaiser
Rabia
R
Bonde
Pramod
P
Duncan
Mark D
MD
Harmon
John W
JW
D000704:Analysis of Variance; D000818:Animals; D002478:Cells, Cultured; D017931:DNA Primers; D004195:Disease Models, Animal; D018274:Electroporation; D051523:Fibroblast Growth Factor 7 / Q000235:genetics* / Q000627:therapeutic use; D015316:Genetic Therapy / Q000379:methods*; D010957:Plasmids / Q000235:genetics / Q000627:therapeutic use; D016133:Polymerase Chain Reaction; D051381:Rats; D017207:Rats, Sprague-Dawley; D018805:Sepsis / Q000503:physiopathology / Q000628:therapy*; D012867:Skin; D014162:Transfection; D014945:Wound Healing / Q000235:genetics* / Q000502:physiology
D017931:DNA Primers; D051523:Fibroblast Growth Factor 7