Success and limitations of a naked plasmid transfection protocol for keratinocyte growth factor-1 to enhance cutaneous wound healing.
Our group and others have previously reported enhancement of cutaneous wound healing following the transfection of tissue with plasmid vectors expressing the DNA for growth factors. In these experiments, growth factor treated animals were usually compared to animals treated with control plasmid vector. To achieve consistent transfection, high DNA plasmid load and repeated penetrations of the wound by needle or gene gun were required. In the current experiments, we assessed the effect of the plasmid load and repeated tissue penetrations on wound healing of excisional wounds in diabetic C57 mice. Animals received 5 mm excisional wounds, and were assigned to the following groups, no treatment, phosphate buffered saline solution injections, and plasmid vector injection with and without the keratinocyte growth factor-1 gene. Intradermal injections of 100 microg plasmid were given adjacent to the wounds at days 1-5, 7 and 11. At day 9, wound closure was more advanced in keratinocyte growth factor-1 treated animals compared to those treated with control plasmid. But a detrimental effect of the DNA plasmid injection was evident from a comparison of the DNA control group versus the non-injected group. Therefore, the challenge for developing an effective system for the enhancement of wound healing lies in improving transfection efficiency.
Byrnes
C K
CK
Section of Surgical Sciences, Johns Hopkins Bayview Medical Center, Johns Hopkins Medical Institutions, Baltimore, Maryland 21224, USA.
Khan
F H
FH
Nass
P H
PH
Hatoum
C
C
Duncan
M D
MD
Harmon
J W
JW
D000818:Animals; D004195:Disease Models, Animal; D005260:Female; D051523:Fibroblast Growth Factor 7; D005346:Fibroblast Growth Factors / Q000235:genetics* / Q000627:therapeutic use*; D005822:Genetic Vectors / Q000235:genetics* / Q000627:therapeutic use*; D051379:Mice; D008810:Mice, Inbred C57BL; D010957:Plasmids / Q000235:genetics* / Q000627:therapeutic use*; D014162:Transfection; D014945:Wound Healing / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D014950:Wounds, Penetrating / Q000188:drug therapy* / Q000235:genetics* / Q000503:physiopathology
C496454:Fgf7 protein, mouse; D051523:Fibroblast Growth Factor 7; D005346:Fibroblast Growth Factors