Dynamic decellularization and cross-linking of rat tracheal matrix

• Published in: Biomaterials 2014-08, Vol.35 (24), p.6344-6350
• Main Authors: Baiguera, Silvia, Del Gaudio, Costantino, Kuevda, Elena, Gonfiotti, Alessandro, Bianco, Alessandra, Macchiarini, Paolo
• Format: Article
• Language: English
• Subjects: Advanced Basic Science; Animals; Biocompatible Materials - pharmacology; Biomechanical Phenomena - drug effects; Bioreactor; Cross-linking; Cross-Linking Reagents - pharmacology; Cytocompatibility; Decellularization; Dentistry; Elastin - metabolism; Extracellular matrix; Extracellular Matrix - drug effects; Extracellular Matrix - metabolism; Extracellular Matrix - ultrastructure; Humans; Iridoids - pharmacology; Male; Materials Testing; Medicin och hälsovetenskap; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - drug effects; Mesenchymal Stromal Cells - ultrastructure; Rats, Inbred BN; Trachea; Trachea - cytology; Trachea - ultrastructure
• ISSN: 0142-9612; 1878-5905; 1878-5905
• DOI: 10.1016/j.biomaterials.2014.04.070

Abstract Decellularized tissues and organs represent a suitable option for tissue engineering when specific scaffolds are needed. However, the optimal conditions to completely remove all the cellular components and minimally affect the biochemical and structural properties of the extracellular matrix are still to be found. For this aim, bioreactors could be an alternative means to dynamically treat the biological samples, automatically controlling all the variables involved in the process and speeding up the entire procedure in order to deal with a suitable scaffold within a limited time period. This paper presents the characterization of rat tracheae decellularized in dynamic conditions, implementing a detergent-enzymatic method, previously considered. Only 6 cycles were enough to generate a tracheal matrix that was histologically and structurally similar to the native one. The network of collagen, reticular and elastic fibers was well preserved, such as the epithelial cilia, the luminal basement membrane and the main matrix components. The elastin content decreased, even if not significantly, after the decellularization protocol. Mechanical properties of the treated tissues were slightly affected by the procedure, and were partially recovered after crosslinking with genipin, a naturally-derived agent. The use of bioreactors could enhance the decellularization procedure of tissues/organs, but a careful selection of the processing parameters is needed in order to prevent large modifications compared to the native condition.