Particle-mediated gene transfer with transforming growth factor-beta1 cDNAs enhances wound repair in rat skin.
Based on preliminary but variable results with direct DNA transfer into wounds, we evaluated in vivo gene transfer by particle-mediated DNA delivery to rat skin to determine whether overexpression of TGF-beta1 at the site of skin incisions would result in a significant improvement in repair. Optimization of the method with viral promoter-luciferase reporter constructs indicated that expression of luciferase activity persisted up to 5 d and was promoter, pressure, and site dependent (ventral > dorsal). Using cytomegalovirus (CMV)-driven human alpha1-antitrypsin, transgene expression was immunolocalized within keratinocytes of the stratum granulosum at 24 h. We measured tensile strength of skin incisions at 11-21 d in both normal and diabetic rats transfected with TGF-beta1 expression vectors at surgery. Native murine TGF-beta1 under an SV40 promoter produced positive effects, while wound strengthening was more pronounced in diabetic animals using a CMV-driven construct. Transfection of rat skin with constitutively active, mutant porcine TGF-beta1 under the control of the CMV and Moloney murine leukemia virus promoters significantly increased tensile strength up to 80% for 14-21 d after surgery. Transfection 24 h before surgery was more effective. Particle-mediated gene delivery can be used to deliver viral promoter-cytokine expression constructs into rat skin in a safe, efficient, and reproducible fashion. The extent of wound repair, as evidenced by enhanced tensile strength, can be markedly improved in tissues transfected with TGF-beta1 expression constructs.
Benn
S I
SI
Department of Pediatrics, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-2561, USA.
Whitsitt
J S
JS
Broadley
K N
KN
Nanney
L B
LB
Perkins
D
D
He
L
L
Patel
M
M
Morgan
J R
JR
Swain
W F
WF
Davidson
J M
JM
D000818:Animals; D019470:Biolistics; D015139:Blotting, Southern; D018076:DNA, Complementary / Q000235:genetics; D003920:Diabetes Mellitus; D005786:Gene Expression Regulation / Q000235:genetics; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D015603:Keratinocytes; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics; D051381:Rats; D012867:Skin / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000494:pharmacology*; D014945:Wound Healing / Q000502:physiology*; D000515:alpha 1-Antitrypsin / Q000235:genetics / Q000494:pharmacology
D018076:DNA, Complementary; D016212:Transforming Growth Factor beta; D000515:alpha 1-Antitrypsin