Development of acellular respiratory mucosal matrix using porcine tracheal mucosa

Background & Aim Respiratory mucosa defect results airway obstruction and infection. Functional recovery of respiratory epithelium, including ciliary movement and mucus secretion, is mandatory. Since site-specific extracellular matrix (ECM) facilitates restoration of the organ function by helping cellular migration and engraftment, a decellularized trachea was considered as ideal ECM in previous studies. However, incomplete removal of cells in cartilage and destruction of mucosal architecture were frequently reported. This results due to the different susceptibility of tissues to various decellularizing agent. To address this issue, the idea of conducting the individual decellularization protocol for separated respiratory mucosa and cartilage was raised. Methods, Results & Conclusion Porcine tracheas were obtained from adult market-sized pig. The trachea was divided into six groups according to their decellularization protocol as follows; native mucosa(N), freezing-thawing(FT), freezing-thawing-DNase/Perasafe™-based chemical agents before separation (tFTD/tFTP) and after separation(mFTD/mFTP). Decelluarization efficacy was evaluated using DNA quantification and H&E stain. ECM contents of the scaffold were evaluated by histological evaluation, glycosaminoglycan(GAG)/collagen assay. Biocompatibility was accessed by cell viability assay and in vivo transplantation. The scaffolds from chemical agent protocols showed low DNA contents ( When comparing the differences by chemical agents, GAG and collagen content was higher in DNase groups than Perasafe™ groups. Laminin and fibronectin were preserved more in mFTP groups. The Cells remained viable when seeded upon the mFTP scaffolds in vitro, and in vivo host response showed a pattern of constructive remodeling when the mFTP scaffolds were implanted. Xenogenic acellular respiratory mucosa matrix which was fabricatied using Perasafe as chemical decellularizing agent on separated porcine tracheal mucosa demonstrated suitable biocompatibility as a scaffold material for respiratory mucosa engineering.