Weymann Alexander, Patil Nikhil Prakash, Sabashnikov Anton, Korkmaz Sevil, Li Shiliang, Soos Pal, Ishtok Roland, Chaimow Nicole, Pätzold Ines, Czerny Natalie, Schmack Bastian, Popov Aron-Frederik, Simon Andre Rüdiger, Karck Matthias, Szabo Gabor
Department of Cardiac Surgery, Heart and Marfan Center, University of Heidelberg, Heidelberg, Germany.
Department of Cardiothoracic Transplantation & Mechanical Circulatory Support, Royal Brompton and Harefield NHS Foundation Trust, Middlesex, UK.
Artif Organs. 2015 Dec;39(12):1024-32. doi: 10.1111/aor.12481. Epub 2015 Apr 20.
Decellularization of native organs may provide an acellular tissue platform for organ regeneration. However, decellularization involves a trade-off between removal of immunogenic cellular elements and preservation of biomechanical integrity. We sought to develop a bioartificial scaffold for respiratory tissue engineering by decellularization of porcine lungs and trachea while preserving organ architecture and vasculature. Lung-trachea preparations from 25 German Landrace pigs were perfused in a modified Langendorff circuit and decellularized by an SDC (sodium deoxycholate)-based perfusion protocol. Decellularization was evaluated by histology and fluorescence microscopy, and residual DNA quantified spectrophotometrically and compared with controls. Airway compliance was evaluated by endotracheal intubation and mechanical ventilation to simulate physiological breathing-induced stretch. Structural integrity was evaluated by bronchoscopy and biomechanical stress/strain analysis by measuring passive tensile strength, all compared with controls. Decellularized lungs and trachea lacked intracellular components but retained specific collagen fibers and elastin. Quantitative DNA analysis demonstrated a significant reduction of DNA compared with controls (32.8 ± 12.4 μg DNA/mg tissue vs. 179.7 ± 35.8 μg DNA/mg tissue, P < 0.05). Lungs and trachea decellularized by our perfusion protocol demonstrated increased airway compliance but preserved biomechanical integrity as compared with native tissue. Whole porcine lungs-tracheae can be successfully decellularized to create an acellular scaffold that preserves extracellular matrix and retains structral integrity and three-dimensional architecture to provide a bioartifical platform for respiratory tissue engineering.
天然器官的去细胞化可为器官再生提供一个无细胞组织平台。然而，去细胞化涉及在去除免疫原性细胞成分与保持生物力学完整性之间进行权衡。我们试图通过对猪肺和气管进行去细胞化处理，同时保留器官结构和脉管系统，来开发一种用于呼吸组织工程的生物人工支架。对来自25头德国长白猪的肺-气管标本在改良的Langendorff循环中进行灌注，并通过基于脱氧胆酸钠（SDC）的灌注方案进行去细胞化处理。通过组织学和荧光显微镜对去细胞化进行评估，用分光光度法定量残留DNA并与对照进行比较。通过气管插管和机械通气来评估气道顺应性，以模拟生理呼吸诱导的拉伸。通过支气管镜评估结构完整性，并通过测量被动抗张强度进行生物力学应力/应变分析，所有这些均与对照进行比较。去细胞化的肺和气管缺乏细胞内成分，但保留了特定的胶原纤维和弹性蛋白。定量DNA分析表明，与对照相比DNA显著减少（32.8±12.4μg DNA/mg组织 vs. 179.7±35.8μg DNA/mg组织，P<0.05）。与天然组织相比，通过我们的灌注方案去细胞化的肺和气管显示出气道顺应性增加，但保留了生物力学完整性。完整的猪肺-气管可以成功地去细胞化，以创建一个保留细胞外基质并保持结构完整性和三维结构的无细胞支架，从而为呼吸组织工程提供一个生物人工平台。
