Lung Tissue Engineering and Preservation of Alveolar Microstructure Using a Novel Casting Method Publisher Pubmed



Kajbafzadeh AM¹ ; Sabetkish N¹ ; Sabetkish S¹ ; Tavangar SM² ; Beigi RSH¹ ; Talebi MA¹ ; Akbarzadeh A¹ ; Nikfarjam L¹ Show Affiliations
Source: Biotechnic and Histochemistry Published:2015

Abstract

We used a rat model to decellularize and seed alveolar cells on a three-dimensional lung scaffold to preserve alveolar microarchitecture. We verifi ed 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 fl uorescent 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 verifi ed 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 confi rmed by SEM of lung casts increases the probability of an effective cell seeding process. © 2014 The Biological Stain Commission.