Abstract
Preventing surgical flaps necrosis remains challenging. Laser Doppler imaging and ultrasound can monitor blood flow in flap regions, but they do not directly measure the cellular response to ischemia. The study aimed to investigate the efficacy of synergistic in-vivo electroporation-mediated gene transfer of interleukin 10 (IL-10) with either hepatocyte growth factor (HGF) or vascular endothelial growth factor (VEGF) on the survival of a modified McFarlane flap, and to evaluate the effect of the treatment on cell metabolism, using label-free fluorescence lifetime imaging. Fifteen male Wistar rats (290–320 g) were randomly divided in three groups: group-A (control group) underwent surgery and received no gene transfer. Group-B received electroporation mediated hIL-10 gene delivery 24 h before and VEGF gene delivery 24 h after surgery. Group-C received electroporation mediated hIL-10 gene delivery 24 h before and hHGF gene delivery 24 h after surgery. The animals were assessed clinically and histologically. In addition, label-free fluorescence lifetime imaging was performed on the flap. Synergistic electroporation mediated gene delivery significantly decreased flap necrosis (P = 0.0079) and increased mean vessel density (P = 0.0079) in treatment groups B and C compared to control group-A. NADH fluorescence lifetime analysis indicated an increase in oxidative phosphorylation in the epidermis of the group-B (P = 0.039) relative to controls. These findings suggested synergistic in-vivo electroporation-mediated gene transfer as a promising therapeutic approach to enhance viability and vascularity of skin flap. Furthermore, the study showed that combinational gene therapy promoted an increase in tissue perfusion and a relative increase in oxidative metabolism within the epithelium.
Highlights
Post-surgical flaps necrosis remains a challenge for the surgeons [1]
We investigated the synergistic effects of in vivo electroporation-mediated gene transfer of interleukin 10 (IL-10) with either vascular endothelial growth factor (VEGF) or hepatocyte growth factor (HGF) on reduction of skin flap necrosis
Combined electroporation mediated IL-10 and HGF gene delivery decreased flap necrosis percentage compared to the control group significantly (Flap necrosis percentage: 25.49 ± 1.65% vs. 35.23 ± 3.90%; p = 0.0079, respectively)
Summary
Post-surgical flaps necrosis remains a challenge for the surgeons [1]. The promising results of recent studies demonstrate the importance of growth factors in aiding tissue repair and regeneration [1, 2]. Application of novel techniques like nonviral gene therapy could further increase the effectiveness by providing sustained therapeutic level of growth factors locally at the skin flap [1, 3]. Previous preclinical studies by others and us have successfully shown promising results of growth factors like Hepatocyte growth factor (HGF), Vascular endothelial growth factor (VEGF), and Interleukin 10 (IL-10) in attenuating graft necrosis [4,5,6, 8, 9]. Prolonged amelioration of acute rejection in rat lung transplant model using IL-10 and HGF has been demonstrated [10]. Their combined effect on skin graft survival, necrosis, and vascularity has not been reported. Through fluorescence lifetime imaging (FLIM), the protein binding status of NADH can be assessed by measuring the time between excitation and emission [13,14,15]
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