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研究生:郭玉宏
研究生(外文):Kuo, Yu-Hung
論文名稱:施加電場對PC-12類神經細胞生長定位與軸突機械性質之影響
論文名稱(外文):The Effect of Applied Electric Field on PC-12 Neuron-like Cell Outgrowth Orientation and Mechanical Properties
指導教授:朱銘祥朱銘祥引用關係林宙晴林宙晴引用關係
指導教授(外文):Ju, Ming-ShaungLin, Chou-Ching
口試委員:褚晴輝胡晉嘉
口試委員(外文):Ching-Hwei ChueJin-Jia Hu
口試日期:2010-07-15
學位類別:碩士
校院名稱:國立成功大學
系所名稱:奈米科技暨微系統工程研究所
學門:工程學門
學類:材料工程學類
論文種類:學術論文
論文出版年:2010
畢業學年度:98
語文別:中文
論文頁數:66
中文關鍵詞:電刺激PC-12類神經細胞免疫螢光染色原子力顯微鏡鈍角錐力學模型
外文關鍵詞:Electrical stimulationPC-12 neuron-like cellImmunofluorescentAtomic force microscopyBlunted-pyramid model
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當神經束受到病變影響產生斷裂時,如果斷裂間隙過大,必須考慮使用神經導管手術進行神經修復。如果可以在神經導管內施加適當之電刺激,一般相信可以使再生神經纖維束更有效地連結。本研究主要目的為探討神經細胞軸突在電場刺激下,對其生長定位與機械性質之影響。
本研究實驗對像為PC-12類神經細胞,方法為針對同一顆細胞做原子力顯微鏡多點定位壓深實驗,並搭配免疫螢光染色。因為為高解析度、低壓深實驗,因此同時考慮原子力顯微鏡真實探針幾何形狀,使用鈍角錐力學模型分析壓深實驗資料,計算出細胞軸突末端視楊氏模數。
實驗結果顯示,施加電場使軸突生長方向逐漸偏向電場方向。視楊氏模數部分,本研究發現細胞軸突末端視楊氏模數與骨架分佈具有正相關性,另外,電場刺激會造成細胞軸突末端骨架重組並導致視楊氏模數增強。當神經細胞受到電場刺激時,會使生長錐上鈣離子濃度增加,造成軸突生長方向偏向負極。另外板足(filopodium)及偽足(lamellpodium)結構上也會受到鈣離子影響,使軸突末端視楊氏模數增加。
The nerve may be fractured due to trauma or disease, if the gap between nerve segments is too far from each other, the use of nerve conduits is often utilized for nerve repair. By applying appropriate electrical stimulation in nerve conduit, it is believed that regeneration of nerve fibers can enhanced. The main purpose of this thesis was to explore outgrowth orientation and mechanical properties of the nerve axons affected by the electrical field stimulation.
The subjects in this study are PC-12 neuron-like cells. Methods for multi-point indentation using atomic force microscope and immunofluorescence image for the same cell. A blunted-pyramid model by considering the real probe geometry was employed to calculate the apparent Young’s modulus of the axon
The results showed that the orientation of axon moves to the cathode progressively. The mechanical properties of axon are highly correlated to the distribution of cytoskeleton. The calcium ion concentration at growth cone may be increased due to electric field and the ultra structure filopodium and lamellpodium might be reorganized and their apparent Young’s modulus were increased.
摘 要 I
Abstract II
致 謝 IV
目 錄 V
圖目錄 VII
表目錄 VIII
符號表 IX
第一章 緒論 1
1.1 研究背景 1
1.2 文獻回顧 5
1.3 研究動機及目的 8
1.4 本文架構 9
第二章 方法與實驗 10
2.1 PC-12類神經細胞培養及軸突誘發程序 10
2.2 原子力顯微鏡 13
2.3 電刺激實驗 17
2.4 免疫螢光染色實驗 20
2.5 PC-12類神經細胞軸突多點壓深實驗 22
2.6 鈍角錐力學模型 25
2.7 實驗流程 33
第三章 實驗結果 35
3.1 PC-12類神經細胞軸突定位結果 35
3.2 PC-12類神經細胞多點壓深實驗結果 41
第四章 討論 52
4-1 PC-12類神經細胞軸突生長機制 52
4-2 PC-12類神經細胞軸突末端(包含生長錐)機械性質 56
4-3 力學模型比較 59
第五章 結論與建議 62
5.1結論 62
5.2建議 63
參 考 文 獻 64
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