Andree C, Swain W F, Page C P, Macklin M D, Slama J, Hatzis D, Eriksson E
Division of Plastic Surgery, Brigham and Women's Hospital, Boston, MA 02115.
Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):12188-92. doi: 10.1073/pnas.91.25.12188.
This report details the transfer of a human epidermal growth factor (hEGF) expression plasmid to porcine partial-thickness wound keratinocytes by particle-mediated DNA transfer (Accell). After gene transfer an external sealed fluid-filled wound chamber was used to protect the wound, provide containment of the exogenous DNA and expressed peptide, and permit sampling of the wound fluid. Analysis of wound fluid for hEGF and total protein, an indicator of reformation of the epithelial barrier, showed that wounds bombarded with the hEGF plasmid exhibited a 190-fold increase in EGF concentration and healed 20% (2.1 days) earlier than the controls. EGF concentrations in wound fluid persisted over the entire 10-day monitored period, decreasing from 200 pg/ml to 25 pg/ml over the first 5 days. Polymerase chain reaction results showed that plasmid DNA was present in the wound for at least 30 days. These findings demonstrate the possible utility of in vivo gene transfer to enhance epidermal repair.
本报告详细介绍了通过粒子介导的DNA转移(Accell)将人表皮生长因子(hEGF)表达质粒转移至猪的部分厚度伤口角质形成细胞的过程。基因转移后,使用外部密封的充满液体的伤口腔室来保护伤口,对外源DNA和表达的肽进行隔离,并允许对伤口液体进行采样。对伤口液体中的hEGF和总蛋白(上皮屏障重塑的指标)进行分析表明,用hEGF质粒轰击的伤口中EGF浓度增加了190倍,并且比对照组提前20%(2.1天)愈合。在整个10天的监测期内,伤口液体中的EGF浓度持续存在,在最初5天内从200 pg/ml降至25 pg/ml。聚合酶链反应结果表明,质粒DNA在伤口中至少存在30天。这些发现证明了体内基因转移在增强表皮修复方面的潜在效用。