Regeneration of partially decellularized tracheal scaffolds in a mouse model of orthotopic tracheal replacement

Abstract

Decellularized tracheal scaffolds offer a potential solution for the repair of long-segment tracheal defects. However, complete decellularization of trachea is complicated by tracheal collapse. We created a partially decellularized tracheal scaffold (DTS) and characterized regeneration in a mouse model of tracheal transplantation. All cell populations except chondrocytes were eliminated from DTS. DTS maintained graft integrity as well as its predominant extracellular matrix (ECM) proteins. We then assessed the performance of DTS in vivo. Grafts formed a functional epithelium by study endpoint (28 days). While initial chondrocyte viability was low, this was found to improve in vivo. We then used atomic force microscopy to quantify micromechanical properties of DTS, demonstrating that orthotopic implantation and graft regeneration lead to the restoration of native tracheal rigidity. We conclude that DTS preserves the cartilage ECM, supports neo-epithelialization, endothelialization and chondrocyte viability, and can serve as a potential solution for long-segment tracheal defects.

Tissue-engineered composite tracheal grafts create mechanically stable and biocompatible airway replacements

“…Partially decellularized tracheal grafts (PDTG) have demonstrated the capacity to support host-derived regeneration of an epithelium and microvasculature 7 – 9 , 34 – 37 while supporting chondrocyte viability. 7 When compared to conventional decellularization approaches that target the complete removal of all cell types, partial decellularization is more effective in preserving graft ECM and associated mechanical properties ( Figure 1 ).…”

“…PDTG were prepared as previously published. 7 Briefly, harvested tracheas were rinsed with 1X PBS with 1% penicillin/streptomycin (P/S, Gibco, Thermo Fisher Scientific, Waltham, MA), then treated with 0.01% (w/v) sodium dodecyl sulfate solution (SDS, Sigma-Aldrich, MO) for 5 min. Tracheas were washed with 0.9% sodium chloride (NaCl, Fisher Scientific, Fair Lawn, NJ) solution three times for progressive 10-, 15-, and 20-min sessions.…”

“… 31 , 32 Immunofluorescent (IF) staining for epithelial (ACT, CCSP, K5/K14) and endothelial (CD31) biomarkers was completed using previously described methods. 7 , 19 …”

“…We demonstrated that partially decellularized tracheal grafts (PDTG) can sustain host-derived epithelialization and endothelialization while supporting graft chondrocyte viability. 7 , 12 , 14 – 16 Despite rapid neotissue formation, one of the predominant complications of any airway reconstruction surgery is stenosis or collapse of the surgically corrected airway. 8 , 11 , 17 , 18 We assessed the performance of composite tracheal grafting, creating a hybrid graft composed of PDTG combined with external splinting.…”

Partial Decellularized Scaffold Combined with Autologous Nasal Epithelial Cell Sheet for Tracheal Tissue Engineering

“…However, with the specific character of the trachea in maintaining the airway structure during inspiration, a completed decellularized scaffold seems to not be an appropriate choice for tracheal replacement, due to lack of a mechanical structure after transplantation [ 5 , 6 ]. Instead, alternative approaches to speed up the process of scaffold generation, including the preservation of the original cartilage cells inside the tracheal scaffold, have been proposed [ 7 , 8 , 9 ]. In our previous study, we introduced a new partial decellularization (PD) protocol that can fabricate a transplantable scaffold with preserved viable cartilage cellular components that remain functional and gradually restore the luminal strength of the tracheal transplant without signs of immune rejection [ 10 ].…”