title: Preservation of micro-architecture and angiogenic potential in a pulmonary acellular matrix obtained using intermittent intra-tracheal flow of detergent enzymatic treatment.
abstract: Tissue engineering of autologous lung tissue aims to become a therapeutic alternative to transplantation. Efforts published so far in creating scaffolds have used harsh decellularization techniques that damage the extracellular matrix (ECM), deplete its components and take up to 5 weeks to perform. The aim of this study was to create a lung natural acellular scaffold using a method that will reduce the time of production and better preserve scaffold architecture and ECM components. Decellularization of rat lungs via the intratracheal route removed most of the nuclear material when compared to the other entry points. An intermittent inflation approach that mimics lung respiration yielded an acellular scaffold in a shorter time with an improved preservation of pulmonary micro-architecture. Electron microscopy demonstrated the maintenance of an intact alveolar network, with no evidence of collapse or tearing. Pulsatile dye injection via the vasculature indicated an intact capillary network in the scaffold. Morphometry analysis demonstrated a significant increase in alveolar fractional volume, with alveolar size analysis confirming that alveolar dimensions were maintained. Biomechanical testing of the scaffolds indicated an increase in resistance and elastance when compared to fresh lungs. Staining and quantification for ECM components showed a presence of collagen, elastin, GAG and laminin. The intratracheal intermittent decellularization methodology could be translated to sheep lungs, demonstrating a preservation of ECM components, alveolar and vascular architecture. Decellularization treatment and methodology preserves lung architecture and ECM whilst reducing the production time to 3 h. Cell seeding and in vivo experiments are necessary to proceed towards clinical translation.
authors.affiliation: Surgery Unit, UCL Institute of Child Health and Great Ormond Street Hospital, 30 Guilford Street, London WC1N 1EH, UK.
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Maghsoudlou
authors.forename: Fanourios
authors.initials: F
authors.lastname: Georgiades
authors.forename: Athanasios
authors.initials: A
authors.lastname: Tyraskis
authors.forename: Giorgia
authors.initials: G
authors.lastname: Totonelli
authors.forename: Stavros P
authors.initials: SP
authors.lastname: Loukogeorgakis
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Orlando
authors.forename: Panicos
authors.initials: P
authors.lastname: Shangaris
authors.forename: Peggy
authors.initials: P
authors.lastname: Lange
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Delalande
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Burns
authors.forename: Angelo
authors.initials: A
authors.lastname: Cenedese
authors.forename: Neil J
authors.initials: NJ
authors.lastname: Sebire
authors.forename: Mark
authors.initials: M
authors.lastname: Turmaine
authors.forename: Brogan N
authors.initials: BN
authors.lastname: Guest
authors.forename: John F
authors.initials: JF
authors.lastname: Alcorn
authors.forename: Anthony
authors.initials: A
authors.lastname: Atala
authors.forename: Martin A
authors.initials: MA
authors.lastname: Birchall
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Elliott
authors.forename: Simon
authors.initials: S
authors.lastname: Eaton
authors.forename: Agostino
authors.initials: A
authors.lastname: Pierro
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Gilbert
authors.forename: Paolo
authors.initials: P
authors.lastname: De Coppi
mesh_terms: D000818:Animals; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000737:chemistry / Q000166:cytology; D005109:Extracellular Matrix / Q000737:chemistry*; D008168:Lung / Q000737:chemistry* / Q000166:cytology; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D018919:Neovascularization, Physiologic / Q000502:physiology; D051381:Rats; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*