Gene gun-delivered human basic fibroblast growth factor gene facilitates the healing of deep partial thickness burn wounds / 中国组织工程研究

ABSTRACT

BACKGROUND: A large amount of in vivo and in vitro experiments have confirmed that, basic fibroblast growth factor (bFGF) has been widely utilized in various tissues and cells, it can facilitate the wound healing.OBJECTIVE: To observe the efficacy and feasibility of gene gun-mediate delivery of human bFGF on the healing of deep partial thickness bum wounds.DESIGN, TIME AND SETTING: Randomized design,an observational trial was performed at the Military Central Laboratory of Changhai Hospital in the Second Military Medical University of Chinese PLA between December 2007 and October 2008.MATERIALS: SD rats of clean grade, weighing 200-250 g, irrespective of genders, ware involved in this study.METHODS: Natural human bFGF gene was recombined and optimized, then eukaryotic expression vector pCI-neo-bFGF was constructed taking pCI-neo as a vector, and transfeoted with human embryonic kidney cells 293 T cells. Dot blot and Western blot methods were utilized to determine the bFGF expression. Rat model of deep partial thickness burn wounds was processed into transgene process using gene gun technique, pCI-neo-bFGF-transfected ones served as experiment group while pCI-neo-transfected ones served as controls.MAIN OUTCOME MEASURES:Wound healing time was recorded and the efficacy was evaluated. The contents of hydroxyproline and collaganase Ⅰ in burn wound tissues were determined at 24 hours, 48 hours, 96 hours, 7 days, 10 days and 14 days following transgene process.RESULTS: the recombinant pCI-neo-bFGF was transfected with human embryonic kidney 293T cells. Dot blot and Western blot analysis have showed that, the constructed pCI-neo-bFGF expression vector could express human bFGF, and the expression of synthesized gene was remarkably higher than that of natural gene under fluorescence microscope; gene gun-mediated transgene experiment have showed that, the wound healing time was (13.00+1.31) days in the experiment group and (14.75±1.28) days in the control group, with significant differences (P＜0.05). The contents of hydroxyproline and collagenase Ⅰ reached a peak at 5 days after the injury, that is 48 hours after transfection, and then gradually decreased and maintained at a certain level. The experiment group had higher hydroxyproline levels compared with control group at different time points (P＜0.05, P＜0.01); the collagenase Ⅰ in the experiment group was notably higher than that in the control group at 48 hours and 96 hours after transfection (P＜0.01).CONCLUSION: Gene gun-mediated delivery of human bFGF can short the time of wound healing, increase the contents of hydroxyproline and collagenase Ⅰ during the healing period, accelerate the healing of deep partial thickness burn wounds.