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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 施博仁 | zh_TW |
| dc.contributor.advisor | Po-Jen Shih | en |
| dc.contributor.author | 廖韋淇 | zh_TW |
| dc.contributor.author | Wei-Qi Liao | en |
| dc.date.accessioned | 2025-09-10T16:32:57Z | - |
| dc.date.available | 2025-09-11 | - |
| dc.date.copyright | 2025-09-10 | - |
| dc.date.issued | 2025 | - |
| dc.date.submitted | 2025-07-22 | - |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/99522 | - |
| dc.description.abstract | 角膜內皮營養不良（Fuchs’ Endothelial Corneal Dystrophy, FECD）為全球常見之角膜內皮疾病，其病理特徵為角膜內皮細胞喪失並引發水腫。然而，受限於角膜捐贈者的嚴重不足，傳統角膜移植治療面臨挑戰。因此，具再生潛力的藥物治療已成為近年研究焦點，其中青光眼用藥Rho-associated protein kinase（ROCK）抑制劑被發現可促進角膜內皮細胞的增殖、遷移與傷口癒合。本研究選擇ROCK抑制劑Y-27632與肌凝蛋白活化劑，處理人類角膜內皮細胞株B4G12，並建立傷口癒合模式進行觀察，探討肌凝蛋白在細胞癒合過程中的功能角色。實驗採用高解析度time-lapse顯微影像與3×3拼接影像技術，輔以開源影像分割模型萃取單細胞形態參數，並開發前後幀影像追蹤演算法以分析細胞在傷口區域的動態行為，進一步結合機器學習方法進行細胞遷移的預測。研究結果顯示，Y-27632可顯著促進體外角膜傷口癒合，此效應主要來自細胞分裂週期的加速及細胞遷移速度的提升，推測其作用機轉與肌凝蛋白活性降低及細胞張力結構重新分布有關。同時也觀察到，鄰近傷口邊緣的細胞運動模式由隨機性逐漸轉變為具方向性的遷移。ROCK抑制對肌凝蛋白的調控亦對整個細胞產生廣泛影響。此外，本研究亦建構一套以影像特徵為基礎的初步預測模型，能有效預測細胞遷移方向，可作為評估細胞修復潛能的輔助工具。整體而言，本研究證實抑制肌凝蛋白活化路徑可有效調控角膜內皮細胞的遷移與癒合，為角膜內皮再生治療策略提供重要的實驗依據與理論支持。 | zh_TW |
| dc.description.abstract | Fuchs’ Endothelial Corneal Dystrophy (FECD) is a common corneal disorder marked by endothelial cell loss and stromal edema. Due to the global shortage of donor corneas, conventional transplantation faces major limitations. As a result, pharmacological therapies with regenerative potential have gained increasing interest. Among them, Rho-associated protein kinase (ROCK) inhibitors—initially used for glaucoma—have shown the ability to enhance corneal endothelial cell proliferation, migration, and wound healing. This study applied the ROCK inhibitor Y-27632 and a myosin activator to human corneal endothelial B4G12 cells in an in vitro wound healing model to investigate myosin’s role in regeneration. High-resolution time-lapse microscopy with 3×3 stitching captured wound-edge dynamics, while an open-source segmentation model extracted single-cell morphological features. A custom tracking algorithm was developed to analyze migration behavior, and machine learning was used to predict migration trajectories. Y-27632 significantly enhanced wound healing by accelerating cell cycle progression and migration speed, likely through reduced myosin activity and redistributed intracellular tension. A shift from random to directional migration was observed near the wound edge, with ROCK inhibition broadly affecting cell behavior. A preliminary image-based model also successfully predicted migration direction, offering a potential tool to assess regenerative capacity. Overall, the findings demonstrate that inhibiting myosin activation effectively modulates corneal endothelial cell migration and wound healing, providing both experimental evidence and theoretical support for regenerative therapies. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2025-09-10T16:32:57Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2025-09-10T16:32:57Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 口試委員會審定書 i
謝誌 ii 中文摘要 iii ABSTRACT iv 目次 v 圖次 ix 表次 xii 第一章 緒論 1 1.1 前言 1 1.2 研究動機 2 第二章 文獻回顧 4 2.1 眼角膜之解剖結構與生理功能 4 2.2 細胞骨架之結構與功能 6 2.3 細胞遷移之機制與調控因子 15 2.4 細胞週期之機制 24 2.5 RhoA–ROCK訊號軸於細胞遷移與週期調控中之角色 29 2.6 角膜內皮功能障礙性疾病之介紹 34 2.7 ROCK Inhibitor作為角膜內皮治療之潛力與應用現況 40 2.8 體外傷口癒合實驗模型 43 2.9 細胞影像分析與機器學習於細胞行為預測之應用 47 第三章 細胞傷口癒合實驗材料方法 52 3.1 實驗規劃流程 52 3.2 細胞培養與實驗所需材料 52 3.2.1. 藥品與耗材 52 3.2.2. 實驗儀器 54 3.2.3. 實驗細胞株 55 3.3 培養人類角膜內皮細胞 56 3.3.1. 細胞繼代 56 3.3.2. 凍細胞 57 3.3.3. 解凍細胞 57 3.3.4. 製備細胞培養基 58 3.4 細胞藥物處理與傷口癒合實驗 58 3.5 細胞免疫螢光染色 59 3.6 實時活細胞影像擷取與多視野拼接 60 第四章 細胞影像分析與資料處理方法 61 4.1 影像分析流程 61 4.2 電腦硬體與軟體環境 61 4.3 傷口區域量測與癒合面積之定量分析 62 4.4 細胞分裂週期之量測與分析 64 4.5 細胞進入分裂之數量於時序上的變化分析 66 4.6 細胞總數於時序上的變化分析 67 4.7 細胞形態學參數於最終時間點之分析 69 4.8 細胞沿傷口中心線法向方向上之空間分布與密度分析 71 4.9 細胞時序追蹤與時空行為動態分析 73 4.10 細胞運動行為預測建立之方法與分析 76 第五章 研究結果與討論 79 5.1 藥物處理對體外傷口癒合之影響 79 5.1.1. 結果 79 5.1.2. 討論 81 5.2 藥物處理對細胞總數增生動態之影響 82 5.2.1. 結果 82 5.2.2. 討論 83 5.3 藥物處理對細胞分裂總數動態之影響 84 5.3.1. 結果 84 5.3.2. 討論 87 5.4 藥物處理對細胞分裂週期之影響 88 5.4.1. 結果 88 5.4.2. 討論 90 5.5 藥物處理下細胞形態學指標之差異 92 5.5.1. 結果 92 5.5.2. 討論 95 5.6 藥物處理對細胞空間分區密度與整體動態之影響 98 5.6.1. 結果 98 5.6.2. 討論 101 5.7 藥物處理對時間與空間分區下細胞動態之影響 102 5.7.1. 結果 102 5.7.2. 討論 111 5.8 基於時序影像的細胞運動行為預測模型 112 5.8.1. 結果 112 5.8.2. 討論 117 5.9 本研究之限制 118 第六章 結論與未來展望 120 6.1 結論 120 6.2 未來展望 122 參考文獻 124 | - |
| dc.language.iso | zh_TW | - |
| dc.subject | 角膜內皮細胞 | zh_TW |
| dc.subject | ROCK抑制劑 | zh_TW |
| dc.subject | AI輔助影像分析 | zh_TW |
| dc.subject | 傷口癒合 | zh_TW |
| dc.subject | 細胞遷移 | zh_TW |
| dc.subject | ROCK inhibitor | en |
| dc.subject | corneal endothelial cells | en |
| dc.subject | cell migration | en |
| dc.subject | AI-assisted image analysis | en |
| dc.subject | wound healing | en |
| dc.title | 人工智慧影像分析肌凝蛋白於角膜內皮癒合之功能 | zh_TW |
| dc.title | AI-Based Image Analysis of Myosin Function in Corneal Endothelial Wound Healing | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 113-2 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 黃義侑;林育君 | zh_TW |
| dc.contributor.oralexamcommittee | Yi-You Huang;Yu-Chun Lin | en |
| dc.subject.keyword | ROCK抑制劑,角膜內皮細胞,傷口癒合,AI輔助影像分析,細胞遷移, | zh_TW |
| dc.subject.keyword | ROCK inhibitor,corneal endothelial cells,wound healing,AI-assisted image analysis,cell migration, | en |
| dc.relation.page | 131 | - |
| dc.identifier.doi | 10.6342/NTU202502010 | - |
| dc.rights.note | 同意授權(限校園內公開) | - |
| dc.date.accepted | 2025-07-23 | - |
| dc.contributor.author-college | 工學院 | - |
| dc.contributor.author-dept | 醫學工程學系 | - |
| dc.date.embargo-lift | 2030-07-21 | - |
| 顯示於系所單位： | 醫學工程學研究所 | |
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|---|---|---|---|
| ntu-113-2.pdf 未授權公開取用 | 7.9 MB | Adobe PDF | 檢視/開啟 |
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