title: Perfusion-Decellularization of Porcine Lung and Trachea for Respiratory Bioengineering.
abstract: 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.
authors.affiliation: 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.
authors.forename: Alexander
authors.initials: A
authors.lastname: Weymann
authors.affiliation: Department of Cardiothoracic Transplantation & Mechanical Circulatory Support, Royal Brompton and Harefield NHS Foundation Trust, Middlesex, UK.
authors.forename: Nikhil Prakash
authors.initials: NP
authors.lastname: Patil
authors.affiliation: Department of Cardiothoracic Transplantation & Mechanical Circulatory Support, Royal Brompton and Harefield NHS Foundation Trust, Middlesex, UK.
authors.forename: Anton
authors.initials: A
authors.lastname: Sabashnikov
authors.affiliation: Department of Cardiac Surgery, Heart and Marfan Center, University of Heidelberg, Heidelberg, Germany.
authors.forename: Sevil
authors.initials: S
authors.lastname: Korkmaz
authors.affiliation: Department of Cardiac Surgery, Heart and Marfan Center, University of Heidelberg, Heidelberg, Germany.
authors.forename: Shiliang
authors.initials: S
authors.lastname: Li
authors.affiliation: Heart and Vascular Center, Semmelweis University, Budapest, Hungary.
authors.forename: Pal
authors.initials: P
authors.lastname: Soos
authors.affiliation: 2nd Department of Pathology, Semmelweis University, Budapest, Hungary.
authors.forename: Roland
authors.initials: R
authors.lastname: Ishtok
authors.affiliation: Department of Cardiac Surgery, Heart and Marfan Center, University of Heidelberg, Heidelberg, Germany.
authors.forename: Nicole
authors.initials: N
authors.lastname: Chaimow
authors.affiliation: Department of Cardiac Surgery, Heart and Marfan Center, University of Heidelberg, Heidelberg, Germany.
authors.forename: Ines
authors.initials: I
authors.lastname: Pätzold
authors.affiliation: Department of Cardiac Surgery, Heart and Marfan Center, University of Heidelberg, Heidelberg, Germany.
authors.forename: Natalie
authors.initials: N
authors.lastname: Czerny
authors.affiliation: Department of Cardiac Surgery, Heart and Marfan Center, University of Heidelberg, Heidelberg, Germany.
authors.forename: Bastian
authors.initials: B
authors.lastname: Schmack
authors.affiliation: Department of Cardiothoracic Transplantation & Mechanical Circulatory Support, Royal Brompton and Harefield NHS Foundation Trust, Middlesex, UK.
authors.forename: Aron-Frederik
authors.initials: AF
authors.lastname: Popov
authors.affiliation: Department of Cardiothoracic Transplantation & Mechanical Circulatory Support, Royal Brompton and Harefield NHS Foundation Trust, Middlesex, UK.
authors.forename: Andre Rüdiger
authors.initials: AR
authors.lastname: Simon
authors.affiliation: Department of Cardiac Surgery, Heart and Marfan Center, University of Heidelberg, Heidelberg, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Karck
authors.affiliation: Department of Cardiac Surgery, Heart and Marfan Center, University of Heidelberg, Heidelberg, Germany.
authors.forename: Gabor
authors.initials: G
authors.lastname: Szabo
mesh_terms: D000818:Animals; D001696:Biomechanical Phenomena; D015153:Blotting, Western; D001999:Bronchoscopy; D004247:DNA / Q000378:metabolism; D003840:Deoxycholic Acid / Q000494:pharmacology*; D005260:Female; D008168:Lung / Q000098:blood supply / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008170:Lung Compliance; D008856:Microscopy, Fluorescence; D010477:Perfusion / Q000379:methods*; D044968:Regenerative Medicine / Q000379:methods*; D012119:Respiration; D012121:Respiration, Artificial; D013053:Spectrophotometry; D013314:Stress, Mechanical; D034421:Sus scrofa; D013718:Tensile Strength; D013997:Time Factors; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D014132:Trachea / Q000098:blood supply / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism
chemical_list: D003840:Deoxycholic Acid; D004247:DNA
keywords: Decellularization; Lung; Tissue engineering; Trachea; Whole organ concept