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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/99576完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳晉興 | zh_TW |
| dc.contributor.advisor | Jin-Shing Chen | en |
| dc.contributor.author | 洪琬婷 | zh_TW |
| dc.contributor.author | Wan-Ting Hung | en |
| dc.date.accessioned | 2025-09-16T16:10:09Z | - |
| dc.date.available | 2025-09-17 | - |
| dc.date.copyright | 2025-09-16 | - |
| dc.date.issued | 2025 | - |
| dc.date.submitted | 2025-07-29 | - |
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Asaio J. 2006;52(1):86–91. 129. Heng WL, Madhavan K, Wee P, Seck T, Lim YP, Lim CH. Banking of cryopreserved iliac artery and vein homografts: clinical uses in transplantation. Cell Tissue Bank. 2015;16(2):235–42. 130. Špaček M, Měřička P, Janoušek L, Štádler P, Adamec M, Vlachovský R, et al. Current vascular allograft procurement, cryopreservation and transplantation techniques in the Czech Republic. Adv Clin Exp Med. 2019;28(4):529–34. 131. Hwang S, Bae JH, Kim IO, Hong JJ. Current vascular allograft procurement, cryopreservation and transplantation techniques in the Asan Medical Center Tissue Bank. Ann Liver Transplant. 2021;1(1):79–85. 132. Capella-Monsonís H, Crum RJ, Hussey GS, Badylak SF. Advances, challenges, and future directions in the clinical translation of ECM biomaterials for regenerative medicine applications. Adv Drug Deliv Rev. 2024;211:115347. | - |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/99576 | - |
| dc.description.abstract | 氣管原生軟骨具有有限的再生能力，使得尋找適合的氣管修補生物材料成為持續的挑戰。本研究探討未經去細胞化之冷凍保存主動脈同種異體移植物作為氣管軟骨再生支架的角色。原本應用於感染性主動脈重建的冷凍保存異體主動脈，具有作為大型動脈的特徵，包括豐富的彈性纖維與平滑肌細胞，並在體外展現出良好的軟骨細胞生物相容性。氣管-冷凍保存異體主動脈修補構造展現出與原生氣管相近的抗拉性能，且能耐受正常呼氣壓力。在兔子的氣管缺損修補模型中，冷凍保存異體主動脈僅引發輕至中度的免疫反應，且反應隨時間逐漸消退。植入一個月內，即可觀察到新生軟骨、血管新生及上皮再生。於後續十二個月中，原有的主動脈支架逐步降解，並由新生的軟骨組織——來源於受贈者的軟骨前驅細胞——取代。蛋白質體分析顯示，冷凍保存異體主動脈富含細胞骨架、細胞黏附、遷移及細胞外基質相關蛋白質。在富含的生長因子中，成纖維細胞生長因子2被確認為促進細胞趨化與軟骨分化的關鍵因子之一；此外，來自冷凍保存異體主動脈的細胞外基質亦展現對軟骨前驅細胞與軟骨細胞的趨觸效應。綜合本研究結果，冷凍保存異體主動脈不僅作為結構支架，同時也作為生物活性平台，透過協同的生物相容性、生長因子訊號傳導及細胞外基質支持，促進氣管軟骨再生。 | zh_TW |
| dc.description.abstract | Native tracheal cartilage exhibits limited regenerative capacity, making the search for suitable biomaterials for tracheal repair a persistent challenge. In this study, a non-decellularized cryopreserved aortic allograft (CAo) is investigated as a scaffold for tracheal cartilage regeneration. Originally used to reconstruct infected aortas, CAo retains key features of a large artery—abundant elastic fibers and smooth muscle cells—and demonstrates favorable in vitro biocompatibility with chondrocytes. A trachea–CAo patch construct maintains tensile properties comparable to native trachea and tolerates normal expiratory forces. In a rabbit patch-defect model, CAo elicits only a mild-to-moderate immune response that gradually subsides. Within one month of implantation, robust neocartilage formation is observed, along with angiogenesis and epithelial regeneration. Over the next 12 months, the original aortic scaffold progressively degrades, while newly formed cartilage—originating from recipient perichondrial chondroprogenitor cells—replaces it. Proteomic analyses show that CAo is enriched in cytoskeletal, adhesion, cell adhesion, migration, and extracellular matrix (ECM)–related proteins. Among the enriched growth factors, fibroblast growth factor 2 (FGF-2) was identified as a key mediator that promotes both chemotaxis and chondrogenic differentiation. In addition, the ECM derived from CAo supports haptotactic migration of chondroprogenitor cells and chondrocytes. These findings indicate that CAo serves as both a structural and biological scaffold, activating tracheal cartilage regeneration through synergistic biocompatibility, growth factor signaling, and ECM support. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2025-09-16T16:10:09Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2025-09-16T16:10:09Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 口試委員會審定書 i
誌 謝 ii 中 文 摘 要 iii Abstract iv Table of Contents v List of Figures ix List of Tables x Chapter 1: Introduction 1 1.1. Clinical challenges in tracheal reconstruction 1 1.2. Current strategies for tracheal reconstruction and their limitations 1 1.3. Emergence of aortic graft in tracheal reconstruction 3 1.4. Clinical Experience of CAo-Based Tracheal Reconstruction at National Taiwan University Hospital 5 1.5. Biological Potential and Challenges of CAo 7 1.6. Research aims and objectives 7 Chapter 2.: Material and Methods 9 2.1. Preparation of cryopreserved aortic allografts (CAo) 9 2.2. Histology Examination 9 2.2.1. Hematoxylin and Eosin (H&E) staining 9 2.2.2. Movat’s pentachrome staining 9 2.2.3. Immunohistochemistry staining 10 2.2.4. Von Kossa staining 11 2.3. Alamar blue assay for aortic tissue metabolic activity 12 2.4. Lactate dehydrogenase (LDH) assay 12 2.5. In vitro aortic tissue culture 12 2.6. Western blot 13 2.7. Primary culture of rabbit tracheal chondrocytes 13 2.8. In vitro biocompatibility test 14 2.9. Mechanical properties 14 2.9.1. Tensile test 14 2.9.2. Blow test of tracheal-CAo patch constructs 15 2.10. Animal experimentation 15 2.11. Laser capture microdissection (LCM), DNA extraction, and PCR analysis 16 2.12. Immunofluorescence (IF) staining for Sox9 and PCNA 17 2.13. Proteomic analysis 17 2.13.1. Protein extraction and digestion 17 2.13.2. LC-MS/MS acquisition 18 2.13.3. Data processing 19 2.13.4. Statistical and bioinformatics analysis 19 2.13.5. Data availability 19 2.14. Primary culture of human airway chondroprogenitor cells (hCPCs) 20 2.14.1. Cell isolation & Fibronectin Adhesion Assay 20 2.14.2. Flow Cytometry of hCPCs 20 2.14.3. Tests of progenitor phenotypic plasticity 21 2.15. Horizontal co-culture 21 2.16. Preparation of CAo-CM 22 2.17. Boyden chamber transwell migration 22 2.18. Haptotaxis assay using decellularized ECM derived from CAo and CTr 23 2.19. Statical analysis 24 Chapter 3: Results 25 3.1. CAo graft characterization 25 3.1.1. Microscopic analyses 25 3.1.2. Metabolic features 25 3.1.3. In vitro biocompatibility with tracheal chondrocyte 26 3.2. Mechanical properties of CAo and trachea-CAo patch constructs 26 3.3. Progressive in vivo cartilage regeneration induced by CAo in tracheal defect repair 27 3.3.1. In vivo biocompatibility 27 3.3.2. Cartilage regeneration and integration 28 3.4. Tracing the origin of regenerated cartilage in CAo scaffolds. 29 3.4.1. Laser capture microdissection and PCR of the neocartilage in gender-mismatch design 29 3.4.2. Evidence of perichondrial involvement in CAo neochondrogenesis 30 3.5. Comparative proteomic analysis of rabbit CAo and tracheal tissues 31 3.6. Validation of effect of FGF-2 released from CAo on chondrogenesis 32 3.7. Validation of haptotactic effect of CAo-derived ECM on chondrocytes and chondroprogenitors 34 Chapter 4: Discussion 36 4.1. Summary of study findings 36 4.2. Distinct tissue remodeling of CAo in tracheal reconstruction compared to vascular replacement 36 4.3. Recipient-derived cell contribution to neocartilage formation in CAo 37 4.4. CAo induces perichondrial chondroprogenitors to initiate chondrogenesis along the scaffold 38 4.5. Proteomic analysis identifies FGF-2 as a key factor driving chondrogenesis within CAo 39 4.6. Angiogenic effects of CAo facilitate tissue regeneration 39 4.7. ECM proteins in CAo promote progenitor adhesion, migration, and matrix deposition 40 4.8. Mechanical limitations of CAo and the evolving rigidity with neocartilage formation 40 4.9. Limitations 41 Chapter 5: Conclusions 42 5.1. Novelty and implication summary 42 5.2. Future perspectives 42 Reference 44 Appendix 55 Appendix 1：Figures 55 Appendix 2：Tables 75 Appendix 3：Related Publications 82 | - |
| dc.language.iso | en | - |
| dc.subject | 氣管 | zh_TW |
| dc.subject | 冷凍保存主動脈 | zh_TW |
| dc.subject | 生物性支架 | zh_TW |
| dc.subject | 軟骨生成 | zh_TW |
| dc.subject | 軟骨膜 | zh_TW |
| dc.subject | 細胞外基質 | zh_TW |
| dc.subject | 成纖維細胞生長因子 | zh_TW |
| dc.subject | extracellular matrices | en |
| dc.subject | chondrogenesis | en |
| dc.subject | biological scaffolds | en |
| dc.subject | tracheae | en |
| dc.subject | cryopreserved aortas | en |
| dc.subject | fibroblast growth factors | en |
| dc.subject | perichondria | en |
| dc.title | 冷凍保存主動脈同種移植物的生物特性及其在氣管軟骨再生中的作用 | zh_TW |
| dc.title | Exploring the Biological Properties of Cryopreserved Aortic Allografts and Their Role in Mediating Tracheal Cartilage Regeneration | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 113-2 | - |
| dc.description.degree | 博士 | - |
| dc.contributor.coadvisor | 許文明;楊台鴻 | zh_TW |
| dc.contributor.coadvisor | Wen-Ming Hsu;Tai-Horng Young | en |
| dc.contributor.oralexamcommittee | 黃凱文;陳祈玲;黃才旺;蔡昕霖 | zh_TW |
| dc.contributor.oralexamcommittee | Kai-Wen Huang;Chi-Ling Chen;Tsai-Wang Huang;Hsin-Lin Tsai | en |
| dc.subject.keyword | 氣管,冷凍保存主動脈,生物性支架,軟骨生成,軟骨膜,細胞外基質,成纖維細胞生長因子, | zh_TW |
| dc.subject.keyword | tracheae,cryopreserved aortas,biological scaffolds,chondrogenesis,perichondria,extracellular matrices,fibroblast growth factors, | en |
| dc.relation.page | 82 | - |
| dc.identifier.doi | 10.6342/NTU202502844 | - |
| dc.rights.note | 同意授權(全球公開) | - |
| dc.date.accepted | 2025-07-30 | - |
| dc.contributor.author-college | 醫學院 | - |
| dc.contributor.author-dept | 臨床醫學研究所 | - |
| dc.date.embargo-lift | 2025-09-17 | - |
| 顯示於系所單位： | 臨床醫學研究所 | |
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