Parikh Punam P, Lassance-Soares Roberta M, Shao Hongwei, Regueiro Manuela M, Li Yan, Liu Zhao-Jun, Velazquez Omaida C
DeWitt-Daughtry Family Department of Surgery, Division of Vascular Surgery, University of Miami Miller School of Medicine, Miami, Florida.
DeWitt-Daughtry Family Department of Surgery, Division of Vascular Surgery, University of Miami Miller School of Medicine, Miami, Florida.
J Surg Res. 2018 Aug;228:68-76. doi: 10.1016/j.jss.2018.02.061. Epub 2018 Mar 26.
Poor wound healing in critical limb ischemia (CLI) is attributed to impaired neovascularization and reperfusion. Optimizing the ischemic wound with adhesion molecules that enhance stem cell homing may revolutionize treatment. The purpose of this study is to test the efficacy of adhesion molecule E-selectin on wound healing in an ischemic mouse wound.
Adult FVB/NJ mice underwent unilateral femoral artery and vein ligation to induce CLI. A 4-mm punch biopsy wound was created on the anterior thigh to simulate ischemic wounds. Intramuscular injection of adeno-associated virus (AAV) carrying either E-selectin (E-selectin/AAV, n = 11) or LacZ as control (LacZ/AAV, n = 10) was performed. Gross wound size was measured for 10 d postoperatively. Ischemic hindlimb reperfusion was quantified using laser Doppler imaging. Wound tissue neovascularization was visualized using DiI perfusion and confocal microscopy. E-selectin expression in wounds was verified by immunofluorescence.
Immunofluorescence confirmed E-selectin/AAV delivery in treatment versus control limbs. Wounds from E-selectin/AAV mice versus controls revealed surface area healing of 54% versus 20% (P < 0.01) on postoperative day (POD) 1, 78% versus 51% on POD 4 (P < 0.01), and 97% versus 84% on POD 10 (P < 0.01). Laser Doppler imaging revealed greater reperfusion in E-selectin/AAV mice versus controls by POD 10 (0.49 versus 0.27, P < 0.05). DiI perfused ligated hindlimb in E-selectin/AAV versus control mice revealed mean neovascularization intensity score of 30 versus 18 (P < 0.05) on POD 10.
Intramuscularly injected E-selectin/AAV gene therapy in mice with CLI significantly increases wound angiogenesis and limb reperfusion, expediting overall wound healing.
严重肢体缺血（CLI）中伤口愈合不良归因于新生血管形成和再灌注受损。用增强干细胞归巢的黏附分子优化缺血伤口可能会彻底改变治疗方法。本研究的目的是测试黏附分子E-选择素对缺血小鼠伤口愈合的疗效。
成年FVB/NJ小鼠接受单侧股动脉和静脉结扎以诱导CLI。在前臂制造一个4毫米的打孔活检伤口以模拟缺血伤口。进行肌肉注射携带E-选择素的腺相关病毒（E-选择素/AAV，n = 11）或作为对照的LacZ（LacZ/AAV，n = 10）。术后10天测量伤口大体尺寸。使用激光多普勒成像对缺血后肢再灌注进行定量。使用DiI灌注和共聚焦显微镜观察伤口组织新生血管形成。通过免疫荧光验证伤口中E-选择素的表达。
免疫荧光证实了治疗组与对照组肢体中E-选择素/AAV的递送。E-选择素/AAV小鼠与对照组的伤口在术后第1天的表面积愈合率分别为54%和20%（P < 0.01），术后第4天分别为78%和51%（P < 0.01），术后第10天分别为97%和84%（P < 0.01）。激光多普勒成像显示，到术后第10天，E-选择素/AAV小鼠的再灌注比对照组更大（0.49对0.27，P < 0.05）。与对照小鼠相比，E-选择素/AAV小鼠中DiI灌注的结扎后肢在术后第10天的平均新生血管形成强度评分为30对18（P < 0.05）。
在CLI小鼠中肌肉注射E-选择素/AAV基因疗法可显著增加伤口血管生成和肢体再灌注，加速整体伤口愈合。
