Primitive regenerated-tissue evolution in situ after implantation of a 3D-printed tracheal graft

Abstract

Tissue-engineered trachea offers a potential solution for patients needing long-segment tracheal resections. In our research, robust neotissue growth, including cartilage, muscle, adipose, and glands, was observed on the external surface of three-dimensional (3D)-printed tracheal grafts transplanted into a 3-month-old large-scale porcine, noticeable after 7 days. This study is the first to categorize the regeneration stages in detail, aiming for a clearer understanding of the cell development process, as previous studies have not fully elucidated the mechanisms of chondrogenesis and glandogenesis. We have identified four stages of chondrogenesis based on chondrocyte numbers, protein expression, and perichondrium presence. Muscle cells evolved from a fibroblast-like state, confirmed by alpha-smooth muscle actin and smooth muscle-myosin heavy chain markers, while initial adipose tissue resembling brown fat diminished over time. Gland development, marked by a change in MUC5B expression, paralleled the findings in native trachea epithelium and submucosal glands. Transforming growth factor-β1 and type XII collagen were key indicators in the emerging neotissue post-transplant.

Keywords

Primitive regenerated-tissue evolution / Vivid neotissue growth / Glandogenesis / Tracheal graft transplantation

Funding

This research was funded by the Ministry of Science and Technology, Taiwan (Grant No. MOST 110-2314-B-075A-013-1); Yen Tjing Ling Medical Foundation, Taiwan (Grant No. CI-112-9); Taichung Veterans General Hospital and National Yang-Ming University, Taiwan (Grant No. TCVGH-YM1100104); Taichung Veterans General Hospital and National Yang-Ming University, Taiwan (Grant No. TCVGH-YM1110105); and Taichung Veterans General Hospital, Taiwan, (Grant No. TCVGH-1104701C).

Conflict of interest

The authors declare they have no competing interests.

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