title: Structural integrity, immunogenicity and biomechanical evaluation of rabbit decelluarized tracheal matrix.
abstract: Decellularization techniques have been widely used as an alternative strategy to produce matrices for organ reconstruction. This study investigated the impact of a detergent-enzymatic decellularization protocol on the extracellular matrix integrity, mechanical properties, and biocompatibility of decellularized tracheal matrices from rabbits. The tracheas of New Zealand white rabbits were decellularized using a modified detergent-enzymatic method (DEM). Antigenicity, cellularity, glycosaminoglycan content, DNA content, histoarchitecture, and mechanical properties were monitored during processing. The surface ultrastructure of the matrix was examined by scanning electron microscopy (SEM). Bioengineered and control tracheas were then implanted in major histocompatibility complex-unmatched rats (xenograft) heterotopically for 7, 15, and 30 days. Structural and functional analysis was performed after transplantation. The results showed that seven cycles of decellularization removed most of the cells and eliminated antigenicity. Histological and molecular biology analysis demonstrated that most of the cellular components and nuclear material were removed. SEM analysis revealed that the decellularized matrices retained the hierarchical structure of the native trachea, and biomechanical tests showed that decellularization did not significantly influence the mechanical properties. Seven, 15 and 30 days after implantation, decreased (p < 0.01) inflammatory reactions were observed in the xenograft models for decellularized matrices compared with control tracheas. No increases in IgM or IgG content were observed in rats that received bioengineered tracheas. In conclusion, this work suggests that seven cycles of the DEM generates a bioengineered rabbit tracheal matrix that is structurally and mechanically similar to native trachea.
authors.affiliation: Department of Cardiothoracic Surgery, College of Clinical Medicine, Yangzhou University, Yangzhou, 225001, Jiangsu Province, China; The Research Center for Translational Medicine, Yangzhou University, Yangzhou, 225001, Jiangsu Province, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Sun
authors.forename: Shu
authors.initials: S
authors.lastname: Pan
authors.forename: Hong-Can
authors.initials: HC
authors.lastname: Shi
authors.forename: Fang-Biao
authors.initials: FB
authors.lastname: Zhang
authors.forename: Wei-Dong
authors.initials: WD
authors.lastname: Zhang
authors.forename: Gang
authors.initials: G
authors.lastname: Ye
authors.forename: Xing-Chen
authors.initials: XC
authors.lastname: Liu
authors.forename: Si-Quan
authors.initials: SQ
authors.lastname: Zhang
authors.forename: Chong-Hao
authors.initials: CH
authors.lastname: Zhong
authors.forename: Xiao-Long
authors.initials: XL
authors.lastname: Yuan
mesh_terms: D000818:Animals; D057005:Bioengineering; D001696:Biomechanical Phenomena; D002645:Chickens; D049033:Chorioallantoic Membrane / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D005109:Extracellular Matrix / Q000276:immunology* / Q000648:ultrastructure; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D006025:Glycosaminoglycans / Q000378:metabolism; D064593:Heterografts; D007150:Immunohistochemistry; D008297:Male; D018919:Neovascularization, Physiologic; D011817:Rabbits; D017207:Rats, Sprague-Dawley; D013194:Staining and Labeling; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D014132:Trachea / Q000033:anatomy & histology* / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure
chemical_list: D006025:Glycosaminoglycans; D016222:Fibroblast Growth Factor 2; D004247:DNA
keywords: decellularization; extracellular matrix; mechanical properties; rabbit trachea; tissue engineering