請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/56206完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 王兆麟(Jaw-Lin Wang) | |
| dc.contributor.author | Wen-Hao Hwang | en |
| dc.contributor.author | 黃文顥 | zh_TW |
| dc.date.accessioned | 2021-06-16T05:18:55Z | - |
| dc.date.available | 2020-08-06 | |
| dc.date.copyright | 2020-08-06 | |
| dc.date.issued | 2020 | |
| dc.date.submitted | 2020-07-27 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/56206 | - |
| dc.description.abstract | 不管是經過氣管手術或者關節受損，軟骨組織修復的速度都非常緩慢。在再生醫學的領域當中，有許多研究都嘗試提高幹細胞的增生速度與軟骨分化速度，希望藉此解決軟骨修復難題，本研究亦為此目標對人類的骨髓間質幹細胞進行研究。 實驗使用自製超音波驅動的壓電刺激系統，探討壓電刺激對幹細胞增生數量、軟骨分化程度以及細胞聚合形態的影響。使用西方墨點法與免疫螢光染色對刺激後的細胞內部phospho-ERK與SOX9蛋白質含量進行分析，並以細胞極性與細胞核β-catenin濃度探討壓電刺激與典型和非典型Wnt訊息傳遞路徑之間的關係。 本研究的結果顯示壓電刺激造成幹細胞內部phospho-ERK提升並有效地加快細胞增生。在LIC與LIC-Pro兩種刺激裝置不同的超音波能量層級之下 (約0.6 mW/cm^2與18 mW/cm^2) 皆促進細胞聚合與軟骨分化，且無需使用軟骨分化培養液。在壓電刺激後活化了細胞中典型Wnt訊息傳遞路徑與非典型Wnt細胞平面極性路徑，使細胞產生極性並依照電場較高的區域聚合，隨著軟骨標記蛋白SOX9的提升，聚合的細胞進行軟骨分化。 骨骼中存在許多膠原蛋白使其具有壓電的特性，然而其與體內軟骨形成的關聯性尚不明確。以本研究中的結果推測，壓電環境很有可能在人體內形成軟骨的過程中起作用。在體內產生壓電效應後，幹細胞向著產生電場的區域遷移與聚合，接著在軟骨分化因子的作用下進行分化。 本研究已經對人類骨髓間質幹細胞進行增生、分化與聚合的分段實驗，但並不確定從細胞低密度時持續培養至軟骨分化，壓電刺激是否能夠整合並貫穿在所有過程。各階段細胞內部其他蛋白質變化以及實際在臨床上的應用為未來主要研究方向。 | zh_TW |
| dc.description.abstract | Cartilage repair is a growing market due to aging population. Among various approaches, mesenchymal stem cells have been considered as a promising solution for cartilage tissue engineering and regeneration as chondrocytes extracted from aging patients cannot proliferate easily. The objective of the study is to investigate the effect of piezoelectric stimulation (PE) on bone marrow-derived human mesenchymal stem cells (hMSCs) with respect to proliferation and differentiation. The results revealed that PE promoted the proliferation of hMSCs possibly through elevation of phospho-ERK and its translocation into nucleus. Furthermore, PE can induce chondrogenesis of hMSCs even without the use of differentiation medium. In particular, PE caused clustering of hMSCs in a pattern highly correlated with the charge distribution on the quartz plate. By analyzing the orientation of primary cilia with respect to cells, we found both ultrasound and piezoelectric can induce cell polarity but required differentiation media to exhibit a visible representation. Although piezoelectricity is abundantly available in bones, the mechanism between piezoelectricity and cartilage formation is unclear. The results in the study suggest that piezoelectricity might play a role in hMSC proliferation and chondrogenesis. Furthermore, our results indicate that piezoelectric stimulation might offer a cheaper and faster way to induce chondrogenesis from hMSCs. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T05:18:55Z (GMT). No. of bitstreams: 1 U0001-2707202017335100.pdf: 8522298 bytes, checksum: fce26f9bff11da127449f40944087a48 (MD5) Previous issue date: 2020 | en |
| dc.description.tableofcontents | 口試委員會審定書 I 致謝 II 摘要 IV Abstract V 目錄 VI 圖目錄 IX 表目錄 XI 第一章 緒論 1 1.1 超音波簡介 1 1.1.1 醫用超音波 1 1.1.2 超音波參數 2 1.2 間質幹細胞簡介 3 1.3 超音波與電流刺激對間質幹細胞的研究 5 1.4 艾爾遜藍染色法 6 1.5 西方墨點法和免疫螢光染色 7 1.5.1 西方墨點法與免疫螢光染色簡介 7 1.5.2 實驗使用抗體 7 1.6 研究目的與動機 12 第二章 材料與方法 13 2.1 細胞培養 13 2.2 超音波刺激系統和超音波驅動的壓電刺激系統 14 2.2.1 LIC刺激載台和LIC-Pro刺激載台 14 2.2.2 裝置參數量測 19 2.3 增生實驗 21 2.3.1 增生實驗－實驗組別 22 2.3.2 增生實驗－實驗流程 22 2.3.3 細胞計數方法 23 2.4 艾爾遜藍染色前導實驗 24 2.5 分化實驗 25 2.5.1 分化實驗－實驗組別 26 2.5.2 分化實驗－實驗流程 26 2.4.3 艾爾遜藍染色與軟骨分化比例分析 27 2.6 聚合實驗 29 2.6.1 聚合實驗－實驗組別 29 2.6.2 聚合實驗－實驗流程 30 2.6.3 雲母粉圖形與COMSOL程式模擬 30 2.7 西方墨點法和免疫螢光染色 31 2.7.1 西方墨點法和免疫螢光染色步驟簡介 31 2.7.2 phospho-ERK核內濃度探討 35 2.7.3 軟骨分化基因SOX9探討 37 2.7.4 Wnt訊息傳遞路徑探討 38 2.8 統計分析 43 第三章 實驗結果與討論 44 3.1 增生實驗 44 3.1.1 細胞增生結果 44 3.1.2 免疫螢光染色phospho-ERK核內濃度探討 45 3.1.3 西方墨點法phospho-ERK 47 3.2 分化實驗 48 3.2.1 艾爾遜藍染色法前導實驗 48 3.2.2 間質幹細胞軟骨分化形態 50 3.2.3 分化實驗結果 51 3.2.4 分化實驗－軟骨分化比例分析 55 3.2.5 軟骨分化基因SOX9探討 58 3.4 聚合實驗 60 3.4.1 細胞聚合形態 60 3.4.2 雲母粉圖形、COMSOL程式模擬與細胞形態比較 63 3.4.3 Wnt訊息傳遞路徑探討 67 第四章 結論 75 第五章 未來展望 75 參考文獻 76 | |
| dc.language.iso | zh-TW | |
| dc.subject | 壓電刺激 | zh_TW |
| dc.subject | 微能量超音波 | zh_TW |
| dc.subject | 軟骨分化 | zh_TW |
| dc.subject | 增生 | zh_TW |
| dc.subject | 人類骨髓間質幹細胞 | zh_TW |
| dc.subject | 聚合 | zh_TW |
| dc.subject | very-low-intensity ultrasound | en |
| dc.subject | proliferation | en |
| dc.subject | bone marrow-derived human mesenchymal stem cells | en |
| dc.subject | piezoelectric stimulation | en |
| dc.subject | aggregation | en |
| dc.subject | chondrogenesis | en |
| dc.title | 壓電刺激對人類間質幹細胞增生、分化與聚合的影響 | zh_TW |
| dc.title | The effect of piezoelectric stimulation on human mesenchymal stem cells: proliferation, differentiation and aggregation | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 108-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 林峯輝(Feng-Huei Lin),劉浩澧(Hao-Li Liu),張至宏(Chih-Hung Chang),陳彥榮(Edward Chern) | |
| dc.subject.keyword | 壓電刺激,微能量超音波,人類骨髓間質幹細胞,增生,軟骨分化,聚合, | zh_TW |
| dc.subject.keyword | piezoelectric stimulation,very-low-intensity ultrasound,bone marrow-derived human mesenchymal stem cells,proliferation,chondrogenesis,aggregation, | en |
| dc.relation.page | 80 | |
| dc.identifier.doi | 10.6342/NTU202001926 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2020-07-28 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 醫學工程學研究所 | zh_TW |
| 顯示於系所單位： | 醫學工程學研究所 | |
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