Marti G, Ferguson M, Wang J, Byrnes C, Dieb R, Qaiser R, Bonde P, Duncan M D, Harmon J W
Section of Surgical Sciences, Johns Hopkins Bayview Medical Center, Johns Hopkins Medical Institutions, Baltimore, MD 21224, USA.
Gene Ther. 2004 Dec;11(24):1780-5. doi: 10.1038/sj.gt.3302383.
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.
我们最近证明，在小鼠伤口愈合模型中，电穿孔可增强转染效果。角质形成细胞生长因子（KGF）是上皮细胞增殖和分化的诱导剂，已被证明在糖尿病患者的伤口中表达不足。我们假设，通过裸DNA注射并随后进行电穿孔将KGF递送至切除伤口，将是一种在糖尿病小鼠模型中增强伤口闭合的新颖且可能有效的方法。ELISA检测证实培养的小鼠细胞中产生了KGF蛋白，RT-PCR检测证实从小鼠采集的皮肤样本中有KGF mRNA。总共32只遗传性糖尿病小鼠在其背部有两个相同的切除伤口，并分为两组，一组接受KGF DNA注射和电穿孔，另一组不接受治疗。到第12天，在KGF和电穿孔处理的情况下，超过90%的伤口愈合，而未处理组为40%。伤口的组织学分析表明，与观察到完整和成熟上皮的处理伤口相比，未处理伤口含有微溃疡，上皮薄或不完整，炎症未消退。综上所述，这些发现表明，在延迟伤口愈合模型中，单次注射质粒编码的KGF DNA并结合电穿孔可改善并加速伤口闭合。
