Electroporation improves transfection efficiency in rat wound healing model

Abstract

Abstract Introduction: We are investigating the use of electroporation in vivo to enhance transfection efficiency and improve wound healing with DNA plasmid expression vectors for growth factors. Methods: For electroporation (EP) transfection efficiency and wound healing we used luciferase and keratinocyte growth factor (KGF) DNA plasmid expression vectors respectively (gWIZ-Lux vector controlled by CMV promoter, Invitrogen, Carlsbad, CA). Animals were electroporated at the site of injection within two minutes of plasmid administration, using a square wave electroporator (ECM 830, BTX Genetronics, San Diego, CA). For luciferase, photon emission was quantified using the IVIS Imaging System (Xenogen Corporation, Alameda, CA). For the KGF experiment, healing 4 cutaneous 8mm punch biopsies was assessed using image analysis software based on NIH image (Scion Image, Frederick, MD). A Sprague Dawley rat sepsis model of wound healing with partial cecal ligation was utilized. We found in previous experiments that wound healing was delayed in septic animals. Results: Single EP enhanced average luciferase expression three-fold compared to plasmid without EP (p 5 ± SEM (photons/sec). Values were recorded as total photon flux from each wound, with 4 wounds per rat. Wound size values represent average ± SEM (pixels). Conclusions: These results encourage us to explore further the possible benefit of electroporation-facilitated transfection of growth factors to improve wound healing.