Rapid preparation of decellularized trachea as a 3D scaffold for organ engineering

Abstract

Objective: To shorten the preparation time of rabbit decellularized tracheal matrix through a modified detergent-enzymatic method with higher concentration of DNase (50 kU/mL), providing an experimental and theoretical basis for clinical decellularization technology. Methods: The control group was a natural trachea, and the experimental group was a tracheal matrix subjected to two and four decellularization cycles. The performance of each group of samples was evaluated by histology and immunohistochemical staining, scanning electron microscopy, biomechanical property testing, inoculation and cytotoxicity tests, and allograft experiments. Results: The results showed that the nuclei of the nonchondral areas of the tracheal stroma were essentially completely removed and MHC-I and MHC-II antigens were removed after two decellularization cycles. Histological staining and scanning electron microscopy showed that the extracellular matrix was retained and the basement membrane was intact. Cell inoculation and proliferation tests confirmed that the acellular tracheal matrix had good biocompatibility, and the proliferation capacity of bone mesenchymal stem cells on the matrix was increased in the experimental group compared with the control group ( p < 0.05). Histological staining and CD68 molecular marker analysis after the allograft experiment showed that the inflammatory response of the acellular tracheal matrix was weak and the infiltration of surrounding macrophages was reduced. Conclusion: A modified detergent-enzymatic method with an increased DNase (50 kU/mL) concentration requires only two cycles (4 days) to obtain a decellularized rabbit tracheal matrix with a short preparation time, good biocompatibility, suitable mechanical properties, and reduced preparation cost.

3D Printed Biomimetic PCL Scaffold as Framework Interspersed With Collagen for Long Segment Tracheal Replacement

“…Developing an ideal tracheal replacement is a great challenge in repairing long-segment tracheal defects (Bogan et al, 2016;Wang et al, 2020). Although tissue engineering technology shows great potential, until now, no substantial breakthrough has been made in creating a biomimetic trachea that mimics the native trachea both structurally and mechanically (Machino et al, 2019).…”

Converging 3D Printing and Electrospinning: Effect of Poly(l‐lactide)/Gelatin Based Short Nanofibers Aerogels on Tracheal Regeneration

“…While tissue‐engineered tracheal scaffolds possess great potential for the replacement of defected tracheal tissues, fabricating a suitable tracheal candidate mimicking the NT structurally and functionally still faces numerous challenges. [ 3,24 ] Fabrication techniques, such as electrospinning and 3D printing can be exploited to fabricate tracheal scaffolds. [ 25 ] However, tracheal scaffolds fabricated from each of these techniques face several limitations; electrospun nanofibers exhibit compact structure, which may impede the diffusion of nutrients and oxygen as well as the infiltration of cellular components and tissues into scaffolds.…”

“…[ 1 ] The tracheal defects are usually caused by tumor, infection, and trauma, which can significantly affect the quality of life of an individual. [ 2,3 ] When the defected region is more than 50% of the total tracheal length in adults or about 30% in pediatric patients, it is almost impossible to be regenerated by the conventional surgical methods, including an end‐to‐end anastomosis. [ 4,5 ] This necessitates the implantation of tracheal substitutes for airway reconstruction.…”

Building Scaffolds for Tubular Tissue Engineering

“…• Sealing of tube can be difficult Vascular Niklason et al, 2001;Shen et al, 2003;L'Heureux et al, 2006;Pricci et al, 2009;Konig et al, 2009;Gauvin et al, 2010;Rayatpisheh et al, 2014;Jung et al, 2015;Gui et al, 2016;Zhao et al, 2018;Wang et al, 2018Intestine Grikscheit et al, 2002, 2004 • High degree of customization and control over scaffold production Xi-Xun et al, 2008;Yang et al, 2009;Neff et al, 2011;Lee et al, 2012Intestine Totonelli et al, 2012Trachea Johnson et al, 2016Butler et al, 2017;Ghorbani et al, 2017;Zhong et al, 2019;Batioglu-Karaaltin et al, 2019;Giraldo-Gomez et al, 2019;Wang et al, 2020 the properties and/or structure of the scaffold. The process of electrospinning, particularly for scaffold production, has been covered extensively in previous reviews (Pham et al, 2006;Rocco et al, 2014).…”

“…In recent years, research has been targeted toward improving in vitro preparation methods such as optimizing decellularization through enzymatic, detergent (Zhong et al, 2019), vacuum-assisted (Butler et al, 2017), and chemicalbased techniques (Batioglu-Karaaltin et al, 2019). For example, enhanced enzymatic approaches have drastically reduced tracheal decellularization time (Giraldo-Gomez et al, 2019;Wang et al, 2020). Importantly, the reduced preparation time had no adverse effects on tracheal ECM structure or biomechanical properties and evaded immunogenic or inflammatory responses when implanted in vivo.…”