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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/97808完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 游佳欣 | zh_TW |
| dc.contributor.advisor | Jiashing Yu | en |
| dc.contributor.author | 曲丹尼 | zh_TW |
| dc.contributor.author | Edgar Daniel Quiñones Pardo | en |
| dc.date.accessioned | 2025-07-17T16:05:38Z | - |
| dc.date.available | 2025-07-18 | - |
| dc.date.copyright | 2025-07-17 | - |
| dc.date.issued | 2025 | - |
| dc.date.submitted | 2025-07-14 | - |
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Electrical Stimulation Increases the Secretion of Cardioprotective Extracellular Vesicles from Cardiac Mesenchymal Stem Cells. Cells 2023, 12 (6). DOI: 10.3390/cells12060875 From NLM. (101) Erwin, N.; Serafim, M. F.; He, M. Enhancing the Cellular Production of Extracellular Vesicles for Developing Therapeutic Applications. Pharm Res 2023, 40 (4), 833-853. DOI: 10.1007/s11095-022-03420-w From NLM. (102) Kowkabany, G.; Bao, Y. Nanoparticle Tracking Analysis: An Effective Tool to Characterize Extracellular Vesicles. Molecules 2024, 29 (19). DOI: 10.3390/molecules29194672 From NLM. (103) Auger, C.; Brunel, A.; Darbas, T.; Akil, H.; Perraud, A.; Bégaud, G.; Bessette, B.; Christou, N.; Verdier, M. Extracellular Vesicle Measurements with Nanoparticle Tracking Analysis: A Different Appreciation of Up and Down Secretion. Int J Mol Sci 2022, 23 (4). DOI: 10.3390/ijms23042310 From NLM. (104) Kang, S. Y.; Lee, E. J.; Byun, J. W.; Han, D.; Choi, Y.; Hwang, D. W.; Lee, D. S. 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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/97808 | - |
| dc.description.abstract | 細胞外囊泡 (EV) 的治療潛力已引起廣泛關注，應用於包括傷口癒合和再生醫學在內的一系列臨床領域。然而，在優化 EV 的產量和治療品質方面仍面臨挑戰。在此背景下，出現了兩種截然不同但又互相補充的策略。第一種策略是透過使用獨特的細胞形態來增強 EV 的產生，例如人類脂肪幹細胞 (hASC) 的細胞球體培養。這些 3D 培養物已被證明能夠顯著增加 EV 的分泌，同時改善所產生 EV 的血管生成特性，這對於傷口癒合至關重要。在糖尿病大鼠模型中的體內研究表明，EV 治療後膠原蛋白生成、上皮再生和血管生成增強，凸顯了該方法的治療前景。第二種策略著重於將生物電子界面 (BEI) 與奈米結構基底結合，以調節細胞行為並增加 EV 的產生。此方法採用基於暈苯的奈米纖維陣列，並以膠原蛋白等生物相容性層進行改質，從而增強細胞黏附和增殖。在這些細胞奈米結構系統中施加持續電刺激，可使永生化骨髓基質細胞 (IBMSC) 的胞外囊泡 (EV) 釋放量顯著增加 300%，且對 EV 大小和細胞活力無不良影響。這些發現凸顯了 BEI 和奈米結構材料在增強 EV 生成的同時保持細胞完整性的潛力。細胞形態和生物電子介面這兩種策略都代表著優化 EV 生成和療效的有希望的途徑。兩者結合，可提供一種強大而多層面的方法來提高 EV 的產量和質量，從而增強其在再生醫學、傷口癒合等領域的應用潛力。 | zh_TW |
| dc.description.abstract | The therapeutic potential of extracellular vesicles (EVs) has garnered significant interest for a range of clinical applications, including wound healing and regenerative medicine. However, challenges persist in optimizing both the yield and therapeutic quality of EVs. In this context, two distinct yet complementary strategies have emerged. The first involves enhancing EV production through the use of unique cellular morphologies, such as cell spheroid cultures of human adipose-derived stem cells (hASCs). These 3D cultures have been shown to significantly increase EV secretion while improving the angiogenic properties of the produced EVs, which are critical for wound healing. In vivo studies in diabetic rat models demonstrated enhanced collagen production, re-epithelization, and angiogenesis following EV treatment, underlining the therapeutic promise of this approach. The second strategy focuses on the integration of bioelectronic interfaces (BEIs) with nanostructured substrates to modulate cell behavior and increase EV production. By employing coronene-based nanofiber arrays and modifying them with biocompatible layers like collagen, this approach promotes stronger cell adhesion and proliferation. The addition of continuous electrical stimulation to these cell-nanostructure systems led to a significant 300% increase in EV release from Immortalized Bone Marrow Stromal Cells (IBMSCs), with no adverse effects on EV size or cell viability.These findings highlight the potential of BEIs and nanostructured materials to enhance EV production while maintaining cell integrity. Both strategies—cellular morphologies and bioelectronic interfaces—represent promising avenues for optimizing EV production and therapeutic efficacy. When combined, they offer a powerful, multifaceted approach to improving the yield and quality of EVs, enhancing their potential for applications in regenerative medicine, wound healing, and beyond. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2025-07-17T16:05:38Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2025-07-17T16:05:38Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | Certificate of Thesis/Dissertation Approval from the Oral Defense Committee……...…I
Acknowledgement and/or Dedication………………………………..…………………II Abstract & Keywords (Chinese)…………………………………………………...…..III Abstract & Keywords (English)………………………………………………………..V Table of Contents…..………………………………………………………………….VII List of Figures…………..…………………………………………………………......XII Chapter 1Introduction………...………………………………………………………….1 1.1 Biointerface Engineering…………...………………………………………………..1 1.1.1 Overview of Biointerface Engineering ………………………………...………..1 1.1.2 Surface chemistry …………………………………………………...…………...2 1.1.3 Tissue engineering scaffolds……………………………………………..….…...4 1.1.4 Three-dimensional Cell Culturing Methods…………………………….………..6 1.1.5 Organic Electrochemical Transistor………………………………………...……7 1.1.6 Biomedical Microelectromechanical systems……………………………....……8 1.2 Wound Healing………………………………………………………...…………….9 1.2.1 Overview of Wound Healing…………………………………………………...9 1.2.2 Challenges of wound healing………………………………………………….10 1.2.3 Factors Affecting Wound Healing…………………………………………….12 1.2.4 Diabetic Wound Healing…………………………………………………...…14 1.2.5 Bioengineering Approaches for Wound Healing……………………………...15 1.3 Extracellular Vesicles……………………………………………………………..16 1.3.1 Overview of Extracellular vesicles……………………………………………16 1.3.2 Mechanisms of EV Biogenesis………………………………………………..18 1.3.3 EV Isolation and Characterization Methods…………………………………..20 1.3.4 Strategies for Extracellular vesicles production………………………………20 1.4 Motivations and Aims………………………………………………………….…22 Chapter 2 Materials and Methods………………………………………………….…24 2.1 Materials……………………………………………………………………….….24 2.2 Equipment………………………………………………………………………...27 2.3 Methods………………………………………………………………….……..…29 2.3.1 Monolayer cell culture…………………………………………………..…….29 2.3.2 Sheet morphology formation ..……..……………………………………….…29 2.3.3 Spheroid morphology formation ………………………………………….......30 2.3.4 Isolation of EVs from different cell morphologies…………………………....31 2.3.5 Physical characterization of EVs……………………………………………...32 2.3.6 RNA and MicroRNA data analysis…………………………………………....32 2.3.7 Scratch assay………………………………………………………….……….33 2.3.8 Cell proliferation…………………………………………………………........33 2.3.9 Tube formation assay………………………………………………………….34 2.3.10 In vivo Wound Healing Experiments……………………………………..…34 2.3.11 Histology, Immunochemistry and Immunofluorescence Analysis…………..35 2.3.12 Fabrication of ITO/PEDOT:PSS pattern…………………………………….35 2.3.13 CR-based NF arrays and device assembly…………………………………..36 2.3.14 Surface modification of CR-based NF arrays…………………………….…37 2.3.15 Morphological characterization…………………………………………..…37 2.3.16 Electrochemical characterization…………………………………….………38 2.3.17 OECT Cell studies………………………………………………………...…38 2.3.18 OECT Extracellular vesicles enhancement experiments…………………….39 2.3.19 Isolation of EVs from OECT……………………………………………...…39 2.3.20 Physical characterization of EVs from OECT………………………………40 2.3.21 Statistical analysis……………………………………………………………40 Chapter 3 Results and Discussion ……………………………………………………41 3.1 Three dimensional cell morphology experiments………………………………...41 3.1.1 Selection of morphology spheroid and size …………………………………..41 3.1.1.1 Cell morphology (spheroid) diameter measurement……..………………...41 3.1.1.2 Nanoparticle Tracking Analysis of cell morphology (spheroid) EVs.……42 3.1.1.3 Bicinchoninic Acid assay of cell morphology (spheroid) EVs…………...45 3.1.1.4 Cell viability effect of EVs from cell morphology (spheroids)…………...46 3.1.2 hASC characterization……………………………………………………...…48 3.1.2.1 Histological examination………………………………………..…………48 3.1.2.2 Scanning Electronic Microscopy………………………………………..…50 3.1.3 Physical characterization of EVs……………………………………………...52 3.1.3.1 Nanoparticle Tracking Analysis of cell morphologies EVs……………….52 3.1.3.2 Bicinchoninic Acid assay of cell morphologies EVs……………………..54 3.1.3.3 Transmission Electron Microscopy of cell morphologies EVs……………57 3.1.4 MicroRNA data analysis………………………………………………………58 3.1.4.1 Expression of miRNAs……………………………………………………58 3.1.4.2 EV biogenesis……………………………………………………………...59 3.1.5 Scratch assay…………………………………………………………………..61 3.1.5.1 Cell migration assay……………………………………………………….61 3.1.6 Cell proliferation………………………………………………………………65 3.1.6.1 Cell viability in endothelial cells…………………………………………..65 3.1.6.2 Cell viability in fibroblast cells……………………………………………66 3.1.7 Tube formation assay………………………………………………………….68 3.1.7.1 Angiogenic effect of EVs………………………………………………….68 3.1.8 In vivo Wound Healing Experiments …………………………………...……70 3.1.8.1 Wound closure rate………………………………………………………..70 3.1.9 Histology, Immunohistochemistry and immunofluorescence Analysis………75 3.1.9.1 Promoted granulation tissue regeneration…………………………………75 3.1.9.2 Collagen deposition………………………………………………………..77 3.1.9.3 Epidermal thickness……………………………………………………….81 3.1.9.4 CD31antibody staining……………………………………………………83 3.1.9.5 H&E Staining ……………………………………………………………..87 3.2 OECT electrical stimulation experiments………………………………………...88 3.2.1 Characterization of OECT ……………………………………………………88 3.2.1.1 SEM characterization……………………………………………………...88 3.2.1.2 Brightfield characterization before and after ES. …………………………90 3.2.1.3 Cyclic voltammetry analysis………………………………………………95 3.2.1.4 Electrochemical impedance spectroscopy spectral analysis………………..97 3.2.1.5 Extracellular vesicles profile………………………………………………99 3.2.2 Physical characterization of EVs post ES……………………………………..101 3.2.2.1 Nanoparticle Tracking Analysis of EVs post ES in MCF-7……………...101 3.2.2.2 Nanoparticle Tracking Analysis of EVs post ES in IBMSC …………….105 3.2.2.3 Bicinchoninic Acid assay of EVs post ES in MCF-7…………………….108 3.2.2.4 Bicinchoninic Acid assay of EVs post ES in IBMSC……………………110 3.2.3 Cell characterization post ES………………………………………………...112 3.2.3.1 Cell viability post ES in MCF-7………………………………………….112 3.2.3.2 Cell viability post ES in IBMSC…………………………………………113 Chapter 4 Conclusions and Future Works……………………………………………115 References…………………………………………………………………………….117 | - |
| dc.language.iso | en | - |
| dc.subject | 有機電化學電晶體（OECT） | zh_TW |
| dc.subject | 糖尿病傷口癒合 | zh_TW |
| dc.subject | 單層形態 | zh_TW |
| dc.subject | 片狀形態 | zh_TW |
| dc.subject | 球狀形態 | zh_TW |
| dc.subject | 細胞外囊泡（EV） | zh_TW |
| dc.subject | 循環腫瘤細胞（CTC） | zh_TW |
| dc.subject | 暈苯奈米纖維（CR-NF） | zh_TW |
| dc.subject | diabetes wound healing | en |
| dc.subject | Coronene nanofibers (CR-NF) | en |
| dc.subject | organic electrochemical transistor (OECT) | en |
| dc.subject | circulating tumor cell (CTC) | en |
| dc.subject | extracellular vesicles (EV) | en |
| dc.subject | spheroid morphology | en |
| dc.subject | sheet morphology | en |
| dc.subject | monolayer morphology | en |
| dc.title | 優化細胞外囊泡生成和治療潛力的多方面策略 | zh_TW |
| dc.title | Multifaceted Strategies for Optimizing Extracellular Vesicle Production and Therapeutic Potential | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 113-2 | - |
| dc.description.degree | 博士 | - |
| dc.contributor.oralexamcommittee | 蕭育生;侯詠德;魏暘;楊凱強;林哲緯 | zh_TW |
| dc.contributor.oralexamcommittee | Yu-Sheng Hsiao;Yung-Te Hou;Yang Wei;Kai-Chiang Yang;Che Wei Lin | en |
| dc.subject.keyword | 暈苯奈米纖維（CR-NF）,有機電化學電晶體（OECT）,循環腫瘤細胞（CTC）,細胞外囊泡（EV）,球狀形態,片狀形態,單層形態,糖尿病傷口癒合, | zh_TW |
| dc.subject.keyword | Coronene nanofibers (CR-NF),organic electrochemical transistor (OECT),circulating tumor cell (CTC),extracellular vesicles (EV),spheroid morphology,sheet morphology,monolayer morphology,diabetes wound healing, | en |
| dc.relation.page | 124 | - |
| dc.identifier.doi | 10.6342/NTU202501701 | - |
| dc.rights.note | 同意授權(全球公開) | - |
| dc.date.accepted | 2025-07-16 | - |
| dc.contributor.author-college | 工學院 | - |
| dc.contributor.author-dept | 永續化學科技國際研究生博士學位學程 | - |
| dc.date.embargo-lift | 2025-07-18 | - |
| 顯示於系所單位： | 永續化學科技國際研究生博士學位學程 | |
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