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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 林峯輝 | zh_TW |
dc.contributor.advisor | Feng-Huei Lin | en |
dc.contributor.author | 陳緻宇 | zh_TW |
dc.contributor.author | Zhi-Yu Chen | en |
dc.date.accessioned | 2023-03-19T22:12:44Z | - |
dc.date.available | 2023-12-29 | - |
dc.date.copyright | 2022-09-30 | - |
dc.date.issued | 2022 | - |
dc.date.submitted | 2002-01-01 | - |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/84474 | - |
dc.description.abstract | 肌腱斷裂之病患於手術縫合後,常伴隨著手術部位周邊組織沾黏。若病患發生嚴重術後周邊組織沾黏之現象,需進行二次手術處理沾黏之組織。而臨床常使用水膠或薄膜於手術部位達抗沾黏效果,雖然具有良好之抗沾黏性質但由於肌腱組織之低細胞及低代謝等性質,且手術部位發炎因子的影響下使得肌腱癒合效果不彰。本研究開發以羧甲基纖維素共聚合白芨多醣體之雙層結構多孔洞抗沾粘膜,直接包覆於手術縫合肌腱外圍,期望能達到抗組織沾黏之效果同時又能促進肌腱修復並降低發炎因子。 本研究成功改良先前實驗室萃取白芨多醣體 (BSP) 之流程,結果能有效提升產率自原本17.1%到23.3%。而將人類肌腱細胞與BSP共培養後,結果顯示白芨多醣體不僅能提升人類肌腱細胞增生能力、遷徙能力以及分泌報外基質之能力。接著,利用BDDE將白芨多醣體與羧甲基纖維素 (CMC) 交聯後製備出雙層結構之多孔膜,物化性分析結果顯示成功將CMC交聯BSP且於體外降解試驗能維持至3周。生物相容性結果顯示材料具有良好生物相容性且無生物毒性。體外細胞試驗證實本研究之CMC/BSP雙層結構膜仍具有提升肌腱細胞增生、遷徙之能力且能降低肌腱細胞中經由LPS所誘導之IL-1β、IL-6等發炎因子。細胞貼附實驗證實材料具低纖維母細胞貼附之特性。動物實驗結果顯示兔子斷裂之阿基里斯腱經手術縫合並包裹本研究雙層結構膜於4周後最大拉力與純縫合手術組相比有顯著提升。透過肌腱組織外觀與HE染色之觀察,包裹本研究膜之組別有明顯減少組織沾黏之情形。且本研究材料能有效降低動物肌腱組織中IL-1β、IL-6與TNF-α發炎因子的表現。 | zh_TW |
dc.description.abstract | Patients who tendon rupture are often associated with postoperative peritendinous adhesion surrounding the surgical site after surgical suture. If the patient has severe adhesion to the surrounding tissue after surgery, a second operation is needed to treat the adhesion tissue. We developed a carboxymethyl cellulose (CMC) bilayer structure porous membrane containing Bletilla Striata Polysaccharide (BSP) and is directly wrapped up of the surgical suture tendon. It is expected to achieve anti-tissue adhesion while promoting tendon repair. This study successfully improved the process of extracting the Bletilla Striata polysaccharide from the previous laboratory study, and the results effectively increased the yield from 17.1% to 23.3%. When the human tenocytes were treated with the BSP, the results showed that the BSP could not only enhance the ability of human tenocytes (HTs) to proliferate, migrate, but also secrete extracellular matrix (ECM). BSP was cross-linked with carboxymethyl cellulose (CMC) by using BDDE to prepare a bilayer structure porous membrane (CX/CXB). The FT-IR, NMR physicochemical analysis results shown that CMC was successfully cross-linked to BSP and the degradation test result shown the CX/CXB bilayer structure porous membrane could maintained last up to 3 weeks in PBS buffer. The biocompatibility results showed that the CX/CXB bilayer structure porous membrane has good biocompatibility and non-cytotoxicity. In vitro cell experiments confirmed that the CX/CXB bilayer structure porous membrane in this study still has the ability to enhance rabbit Achilles tenocytes proliferation and migration, and could reduce inflammatory factors such as IL-1β and IL-6 induced by LPS in rabbit Achilles tenocytes. Cell attachment experiments confirmed that the outer-layer CX/CXB bilayer structure porous membrane has low fibroblast attachment properties. The results of animal experiments shown that the maximum tensile force of the ruptured Achilles tendon in rabbits was significantly increased compared with the suture group after 4 weeks of surgical suturing and wrapping the CX/CXB bilayer structure porous membrane in this study. Through the observation of the appearance of tendon tissue and HE staining results, the group wrapped with the CX/CXB bilayer structure porous membrane in this study has significantly reduced tissue adhesion. The results of trichrome stain and inflammatory factor detection experiments shown that the membrane in this study could effectively reduce the expression of inflammatory factors such as IL-1β, IL-6, TNF-α in tendon tissue. | en |
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dc.description.tableofcontents | 口試委員審定書 i 序言 ii 中文摘要 iii ABSTRACT iv CONTENTS vi LIST OF FIGURES x LIST OF TABLES xiv LIST OF ABBREVIATION xv Chapter 1 INTRODUCTION 1 1.1 Epidemiology of Hand Injuries 1 1.2 Statement of the Problem 2 1.2.1 Postoperative adhesion (PA) 2 1.2.2 Inefficient and slow of repair process 3 1.3 Literature Review 4 1.3.1 Anti-adhesion strategies 4 1.3.2 Promote tendon healing 5 1.4 Purpose of Study 6 Chapter 2 THEORETICAL BASIS 7 2.1 Anatomy of Tendon 7 2.2 Tendon Healing 8 2.3 Commercial anti-tissue adhesion products 9 2.3.1 Seprafilm™ 9 2.3.2 ADEPT® 10 2.4 Inflammatory response 11 2.5 Bletilla Striata 12 2.6 Carboxymethyl cellulose 14 2.7 1,4-Butanediol diglycidyl ether (BDDE) 15 Chapter 3 MATERIALS AND METHODS 16 3.1 Experimental Instrument 16 3.2 Experimental Chemicals 17 3.3 Experimental Processes 20 3.3.1 The Effects of Bletilla Striata Polysaccharide (BSP) on the Proliferation and Migration of Tenocytes in vitro 20 3.3.2 Development of Carboxymethyl Cellulose and Bletilla Striata Polysaccharide bilayer structure porous membrane as a Barrier for Preventing Postoperative Peritendinous Adhesion 21 3.4 Extraction of Polysaccharide from Bletilla striata 22 3.5 Isolation and Culture of HTs and RATs 22 3.6 Specific Gene Expression of Human Tenocytes 23 3.7 Immunocytochemical Tenomodulin Staining 24 3.8 Cell Viability of BSP 24 3.9 LIVE/DEAD Staining 25 3.10 Cell Number Calculation 25 3.11 Western Blot (WB) Analysis of Erk and Akt Pathway 25 3.12 Gap Closure Migration Analysis 27 3.13 Transwell Migration Analysis 27 3.14 ECM (Total Collagen) Analysis 28 3.15 Synthesis of Porous Bilayer Structure Membrane 28 3.16 Degradation Rate of Membrane 29 3.17 Swelling Ratio (SR) of Membrane 29 3.18 BSP Release Profile of Membrane 30 3.19 Biocompatibility of Membranes 30 3.20 Proliferation Analysis of Membranes 31 3.21 Attraction Analysis of Membranes 31 3.22 Gene Expression of Inflammatory Cytokines in RATs 32 3.23 Cell Attachment Analysis of Membranes 33 3.24 Animal study 34 3.25 Statistical Analysis 35 Chapter 4 RESULTS AND DISCUSSIONS 36 4.1 The Effects of BSP on the Proliferation and Migration of Tenocytes in vitro 36 4.1.1 Characterization of BSP 36 4.1.2 Biocompatibility of BSP 39 4.1.3 Identification of HTs 40 4.1.4 Cell viability of HTs treated with BSP 42 4.1.5 Cell number of HTs treated with BSP 42 4.1.6 WB analysis of cell proliferation pathway 43 4.1.7 Gap closure migration 46 4.1.8 Cell attracting test 47 4.1.9 ECM synthesis of HTs treated with BSP 49 4.2 Development of CMC and BSP bilayer structure porous membrane as a Barrier for Preventing Postoperative Peritendinous Adhesion 51 4.2.1 Morphology of CX/CXB membrane 51 4.2.2 FT-IR of BDDE crosslinked CMC-BSP membrane 52 4.2.3 NMR analysis of BDDE crosslinked CMC-BSP membrane 52 4.2.4 Degradation of CX/CXB membrane 53 4.2.5 BSP release profile of CX/CXB membrane 54 4.2.6 Swelling ratio analysis of CX/CXB membrane 54 4.2.7 Biocompatibility analysis of CX/CXB membrane 56 4.2.8 Proliferation analysis 57 4.2.9 Migration analysis 58 4.2.10 Gene expression of inflammatory cytokines in RATs 58 4.2.11 Cell attachment test of CX/CXB membrane 59 4.2.12 Animal study 60 4.2.13 Breaking force test of tendon 62 4.2.14 Histological of tendon 63 4.2.15 Inflammatory cytokines expression of tendon tissue 64 4.2.16 ECM analysis of tendon tissue 65 Chapter 5 CONCLUSION 68 5.1 The Effects of BSP on the Proliferation and Migration of Tenocytes in vitro 68 5.2 Development of CMC and BSP Bilayer Structure Porous Membrane as a Barrier for Preventing Postoperative Peritendinous Adhesion 68 REFERENCES 70 | - |
dc.language.iso | en | - |
dc.title | 探討羧甲基纖維素薄膜含白芨多醣體應用於肌腱修復 | zh_TW |
dc.title | Investigation of Carboxymethyl Cellulose Membrane containing Bletilla Striata Polysaccharide for Tendon Healing | en |
dc.type | Thesis | - |
dc.date.schoolyear | 110-2 | - |
dc.description.degree | 博士 | - |
dc.contributor.oralexamcommittee | 黃義侑;郭士民;姚俊旭;陳思恒 | zh_TW |
dc.contributor.oralexamcommittee | Yi-You Huang;Shyh-Ming Kuo;Chun-Hsu Yao;Shih-Heng Chen | en |
dc.subject.keyword | 肌腱縫合手術,術後週邊組織沾黏,白芨多醣體,抗沾粘膜,羧甲基纖維素, | zh_TW |
dc.subject.keyword | tendon surgical suture,postoperative peritendinous adhesion,Bletilla Striata polysaccharide,carboxymethyl cellulose,anti-tissue adhesion membrane, | en |
dc.relation.page | 76 | - |
dc.identifier.doi | 10.6342/NTU202203961 | - |
dc.rights.note | 同意授權(限校園內公開) | - |
dc.date.accepted | 2022-09-26 | - |
dc.contributor.author-college | 工學院 | - |
dc.contributor.author-dept | 醫學工程學系 | - |
dc.date.embargo-lift | 2027-09-24 | - |
顯示於系所單位: | 醫學工程學研究所 |
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