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Herein, a modular strategy for scaffold\u2010free tracheal engineering is developed. A cartilage sheet (Cart\u2010S) prepared by high\u2010density culture is laminated and reshaped to construct a cartilage tube as the main load\u2010bearing structure in which the chondrocytes exhibit a stable phenotype and secreted considerable cartilage\u2010specific matrix, presenting a native\u2010like grid arrangement. To further build a tracheal epithelial barrier, a temperature\u2010sensitive technique is used to construct the monolayer epithelium sheet (Epi\u2010S), in which the airway epithelial cells present integrated tight junctions, good transepithelial electrical resistance, and favorable ciliary differentiation capability. Epi\u2010S can be integrally transferred to inner wall of cartilage tube, forming a scaffold\u2010free complex tracheal substitute (SC\u2010trachea). Interestingly, when Epi\u2010S is attached to the cartilage surface, epithelium\u2010specific gene expression is significantly enhanced. SC\u2010trachea establishes abundant blood supply via heterotopic vascularization and then is pedicle transplanted for tracheal reconstruction, achieving 83.3% survival outcomes in rabbit models. Notably, the scaffold\u2010free engineered trachea simultaneously satisfies sufficient mechanical properties and barrier function due to its matrix\u2010rich cartilage structure and well\u2010differentiated ciliated epithelium, demonstrating great clinical potential for long\u2010segmental tracheal reconstruction.<\/jats:p>","DOI":"10.1002\/adhm.202202022","type":"journal-article","created":{"date-parts":[[2022,12,2]],"date-time":"2022-12-02T23:20:50Z","timestamp":1670023250000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Scaffold\u2010Free Tracheal Engineering via a Modular Strategy Based on Cartilage and Epithelium Sheets"],"prefix":"10.1002","volume":"12","author":[{"given":"Minglei","family":"Yang","sequence":"first","affiliation":[{"name":"Department of Cardiothoracic Surgery  Ningbo No.2 Hospital Ningbo Zhejiang 315000 China"},{"name":"Ningbo Institute of Life and Health Industry University of Chinese 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