title: Production and assessment of decellularized pig and human lung scaffolds.
abstract: The authors have previously shown that acellular (AC) trachea-lung scaffolds can (1) be produced from natural rat lungs, (2) retain critical components of the extracellular matrix (ECM) such as collagen-1 and elastin, and (3) be used to produce lung tissue after recellularization with murine embryonic stem cells. The aim of this study was to produce large (porcine or human) AC lung scaffolds to determine the feasibility of producing scaffolds with potential clinical applicability. We report here the first attempt to produce AC pig or human trachea-lung scaffold. Using a combination of freezing and sodium dodecyl sulfate washes, pig trachea-lungs and human trachea-lungs were decellularized. Once decellularization was complete we evaluated the structural integrity of the AC lung scaffolds using bronchoscopy, multiphoton microscopy (MPM), assessment of the ECM utilizing immunocytochemistry and evaluation of mechanics through the use of pulmonary function tests (PFTs). Immunocytochemistry indicated that there was loss of collagen type IV and laminin in the AC lung scaffold, but retention of collagen-1, elastin, and fibronectin in some regions. MPM scoring was also used to examine the AC lung scaffold ECM structure and to evaluate the amount of collagen I in normal and AC lung. MPM was used to examine the physical arrangement of collagen-1 and elastin in the pleura, distal lung, lung borders, and trachea or bronchi. MPM and bronchoscopy of trachea and lung tissues showed that no cells or cell debris remained in the AC scaffolds. PFT measurements of the trachea-lungs showed no relevant differences in peak pressure, dynamic or static compliance, and a nonrestricted flow pattern in AC compared to normal lungs. Although there were changes in content of collagen I and elastin this did not affect the mechanics of lung function as evidenced by normal PFT values. When repopulated with a variety of stem or adult cells including human adult primary alveolar epithelial type II cells both pig and human AC scaffolds supported cell attachment and cell viability. Examination of scaffolds produced using a variety of detergents indicated that detergent choice influenced human immune response in terms of T cell activation and chemokine production.
authors: Department of Internal Medicine, University of Texas Medical Branch, Galveston, Texas 66555-0435, USA. jnichols@utmb.edu
authors: Joan E
authors: JE
authors: Nichols
authors: Jean
authors: J
authors: Niles
authors: Michael
authors: M
authors: Riddle
authors: Gracie
authors: G
authors: Vargas
authors: Tuya
authors: T
authors: Schilagard
authors: Liang
authors: L
authors: Ma
authors: Kert
authors: K
authors: Edward
authors: Saverio
authors: S
authors: La Francesca
authors: Jason
authors: J
authors: Sakamoto
authors: Stephanie
authors: S
authors: Vega
authors: Marie
authors: M
authors: Ogadegbe
authors: Ronald
authors: R
authors: Mlcak
authors: Donald
authors: D
authors: Deyo
authors: Lee
authors: L
authors: Woodson
authors: Christopher
authors: C
authors: McQuitty
authors: Scott
authors: S
authors: Lick
authors: Daniel
authors: D
authors: Beckles
authors: Esther
authors: E
authors: Melo
authors: Joaquin
authors: J
authors: Cortiella
mesh_terms: D000818:Animals; D003094:Collagen / Q000737:chemistry; D006801:Humans; D007150:Immunohistochemistry; D007797:Laminin / Q000737:chemistry; D008168:Lung; D013552:Swine; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D007797:Laminin; D003094:Collagen
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