3D perfusable minichannels stented with matrix bound vesicles derived from gingival mesenchymal stem cells ameliorated granuloma-related stenosis and improved survival in a rabbit tracheal replacement model.

Abstract

Successful tracheal regeneration remains a significant challenge in tracheal replacement, with tracheal stenosis posing a major obstacle. Decellularized extracellular matrices (dECMs) are emerging as promising alternatives for tissue engineering applications related to the trachea. However, preserving the bioactivity of ECM during the decellularization process presents a significant challenge that compromises its utility in tissue repair, particularly for tracheas that are continuously exposed to pollutants. To address these limitations, we developed a perfusable minichannel scaffold enriched with cartilaginous acellular extracellular matrix-bound vesicles derived from gingival mesenchymal stem cells (gMVs-cACM). Through controlled porosity optimization and the integration of melt electrospinning writing (MEW), this novel perfusable minichannel scaffold facilitates the oriented deposition of the extracellular matrix and the preservation of gMVs via the perfusion decellularization process. In vitro and in vivo functional analyses demonstrated that gMV-cACM generated from perfused decellularization exhibits potent immunomodulatory effects. This was evidenced by tracheal transplantation models, which showed a 54.0 ± 8 % reduction in postoperative granuloma-related stenosis compared to cACM generated from dynamic decellularization, along with an improved survival rate. Mechanistically, proteomic sequencing revealed the downregulation of Interleukin-1β (IL-1β) in response to modulation of the immune microenvironment. In conclusion, this study presents a novel perfusable minichannel scaffold that enhances the bioactivity of dECMs and highlights the potential of gMV-cACM-based scaffolds to advance the field of tracheal tissue engineering.