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      Electroporative transfection with KGF-1 DNA improves wound healing in a diabetic mouse model.

      Gene Therapy

      Animals, Cells, Cultured, Diabetes Mellitus, Experimental, pathology, physiopathology, therapy, Electroporation, Female, Fibroblast Growth Factor 7, Fibroblast Growth Factors, biosynthesis, genetics, Gene Expression, Genetic Therapy, methods, Mice, Mice, Inbred BALB C, Plasmids, RNA, Messenger, Reverse Transcriptase Polymerase Chain Reaction, Skin, metabolism, Transfection, Wound Healing

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          We recently demonstrated that electroporation enhances transfection in a mouse wound-healing model. Keratinocyte growth factor (KGF) is an inducer of epithelial cell proliferation and differentiation and has been shown to be under expressed in the wounds of diabetic individuals. We hypothesized that KGF delivered into an excisional wound via naked DNA injection with subsequent electroporation would be a novel and potentially effective method to enhance wound closure in a diabetic mouse model. ELISA assays confirmed production of KGF protein in cultured mouse cells and RT-PCR assays confirmed KGF mRNA in skin samples taken from mice. In all, 32 genetically diabetic mice were given two identical excisional wounds of their dorsum and split into two groups with one group receiving KGF DNA injection and electroporation with the other group receiving no treatment. Over 90% of wounds healed in the presence of KGF and electroporation versus 40% in the untreated group by day 12. Histological analysis of the wounds demonstrated that untreated wounds contained microulcers with thin or incomplete epithelium with unresolved inflammation as compared to treated wounds where intact and mature epithelium was observed. Taken together these findings suggest that a single injection of KGF DNA encoded on a plasmid coupled with electroporation improves and accelerates wound closure in a delayed wound-healing model.

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