Harder Yves, Schmauss Daniel, Wettstein Reto, Egaña José T, Weiss Fabian, Weinzierl Andrea, Schuldt Anna, Machens Hans-Günther, Menger Michael D, Rezaeian Farid
Department of Plastic Surgery and Hand Surgery, Klinikum rechts der Isar, Technische Universität München;
Department of Plastic Surgery and Hand Surgery, Klinikum rechts der Isar, Technische Universität München.
J Vis Exp. 2014 Nov 17(93):e51900. doi: 10.3791/51900.
Despite profound expertise and advanced surgical techniques, ischemia-induced complications ranging from wound breakdown to extensive tissue necrosis are still occurring, particularly in reconstructive flap surgery. Multiple experimental flap models have been developed to analyze underlying causes and mechanisms and to investigate treatment strategies to prevent ischemic complications. The limiting factor of most models is the lacking possibility to directly and repetitively visualize microvascular architecture and hemodynamics. The goal of the protocol was to present a well-established mouse model affiliating these before mentioned lacking elements. Harder et al. have developed a model of a musculocutaneous flap with a random perfusion pattern that undergoes acute persistent ischemia and results in ~50% necrosis after 10 days if kept untreated. With the aid of intravital epi-fluorescence microscopy, this chamber model allows repetitive visualization of morphology and hemodynamics in different regions of interest over time. Associated processes such as apoptosis, inflammation, microvascular leakage and angiogenesis can be investigated and correlated to immunohistochemical and molecular protein assays. To date, the model has proven feasibility and reproducibility in several published experimental studies investigating the effect of pre-, peri- and postconditioning of ischemically challenged tissue.
尽管拥有深厚的专业知识和先进的手术技术,但缺血引起的并发症,从伤口破裂到广泛的组织坏死,仍在发生,尤其是在重建皮瓣手术中。已经开发了多种实验性皮瓣模型来分析潜在原因和机制,并研究预防缺血性并发症的治疗策略。大多数模型的限制因素是缺乏直接和重复可视化微血管结构和血流动力学的可能性。该方案的目标是提出一个完善的小鼠模型,具备上述缺失的要素。哈德等人开发了一种具有随机灌注模式的肌皮瓣模型,该模型经历急性持续性缺血,如果不进行治疗,10天后会导致约50%的坏死。借助活体荧光显微镜,这种腔室模型可以随着时间的推移对不同感兴趣区域的形态和血流动力学进行重复可视化。可以研究诸如细胞凋亡、炎症、微血管渗漏和血管生成等相关过程,并将其与免疫组织化学和分子蛋白检测相关联。迄今为止,在几项已发表的实验研究中,该模型已证明在研究缺血挑战组织的预处理、围手术期处理和后处理效果方面具有可行性和可重复性。