Particle-mediated gene transfer with transforming growth factor-beta1 cDNAs enhances wound repair in rat skin.

Abstract

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.

• Authors: S I Benn, J S Whitsitt, K N Broadley, L B Nanney, D Perkins, L He, M Patel, J R Morgan, W F Swain, J M Davidson
• Journal: The Journal of clinical investigation (J Clin Invest) Vol. 98 Issue 12 Pg. 2894-902 (Dec 15 1996) ISSN: 0021-9738 [Print] United States
• PMID: 8981938 (Publication Type: Journal Article, Research Support, U.S. Gov't, Non-P.H.S., Research Support, U.S. Gov't, P.H.S.)

Chemical References

• DNA, Complementary
• Transforming Growth Factor beta
• alpha 1-Antitrypsin

Topics

• Animals
• Biolistics
• Blotting, Southern
• DNA, Complementary (genetics)
• Diabetes Mellitus
• Gene Expression Regulation (genetics)
• Gene Transfer Techniques
• Genes, Reporter (genetics)
• HeLa Cells
• Humans
• Immunohistochemistry
• Keratinocytes
• Polymerase Chain Reaction
• Promoter Regions, Genetic (genetics)
• Rats
• Skin (metabolism)
• Transforming Growth Factor beta (genetics, pharmacology)
• Wound Healing (physiology)
• alpha 1-Antitrypsin (genetics, pharmacology)