title: Lung tissue engineering and preservation of alveolar microstructure using a novel casting method.
abstract: We used a rat model to decellularize and seed alveolar cells on a three-dimensional lung scaffold to preserve alveolar microarchitecture. We verified the preservation of terminal respiratory structure by casting and by scanning electron microscopy (SEM) of the casts after decellularization. Whole lungs were obtained from 12 healthy Sprague-Dawley rats, cannulated through the trachea under sterile conditions, and decellularized using a detergent-based method. Casting of both natural and decellularized lungs was performed to verify preservation of the inner microstructure of scaffolds for further cell seeding. Alveolar cell seeding was performed using green fluorescent protein (GFP) lung cells and non-GFP lung cells, and a peristaltic pump. We assessed cell seeding using histological and immunohistochemical staining, and enzymatic evaluation. All cellular components were removed completely from the scaffolds, and histological staining and SEM of casts were used to verify the preservation of tissue structure. Tensile tests verified conservation of biomechanical properties. The hydroxyproline content of decellularized lungs was similar to native lung. Histological and immunohistochemical evaluations showed effective cell seeding on decellularized matrices. Enzymatic measurement of trypsin and alpha 1 antitrypsin suggested the potential functional properties of the regenerated lungs. Casts produced by our method have satisfactory geometrical properties for further cell seeding of lung scaffolds. Preservation of micro-architecture and terminal alveoli that was confirmed by SEM of lung casts increases the probability of an effective cell seeding process.
authors.affiliation: Pediatric Urology Research Center, Section of Tissue Engineering and Stem Cells Therapy, Children's Hospital Medical Center, Tehran University of Medical Sciences , Tehran , Iran.
authors.forename: A-M
authors.initials: AM
authors.lastname: Kajbafzadeh
authors.forename: N
authors.initials: N
authors.lastname: Sabetkish
authors.forename: S
authors.initials: S
authors.lastname: Sabetkish
authors.forename: S M
authors.initials: SM
authors.lastname: Tavangar
authors.forename: R S
authors.initials: RS
authors.lastname: Hossein Beigi
authors.forename: M A
authors.initials: MA
authors.lastname: Talebi
authors.forename: A
authors.initials: A
authors.lastname: Akbarzadeh
authors.forename: L
authors.initials: L
authors.lastname: Nikfarjam
mesh_terms: D000818:Animals; D003902:Detergents; D005109:Extracellular Matrix / Q000648:ultrastructure; D008168:Lung / Q000648:ultrastructure*; D008855:Microscopy, Electron, Scanning; D009926:Organ Preservation / Q000295:instrumentation / Q000379:methods*; D017207:Rats, Sprague-Dawley; D023822:Tissue Engineering / Q000379:methods
chemical_list: D003902:Detergents
keywords: CT scanning; GFP; MRI; bioscaffold; casts; decellularization; hydroxyproline; imaging; lung; scanning electron microscopy; tissue engineering