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系統識別號 U0026-2608201908584400
論文名稱(中文) 以黏彈性有限元素法分析紫杉醇臨床施打計劃對活體PC-12類神經細胞之影響
論文名稱(英文) Effects of Clinical Dosing Schedule of Paclitaxel on Viscoelasticity of Living PC-12 Cells – Finite Element Analysis
校院名稱 成功大學
系所名稱(中) 機械工程學系
系所名稱(英) Department of Mechanical Engineering
學年度 107
學期 2
出版年 108
研究生(中文) 江柏宏
研究生(英文) Bo-Hing Jiang
學號 N16054988
學位類別 碩士
語文別 中文
論文頁數 134頁
口試委員 指導教授-朱銘祥
口試委員-林宙晴
口試委員-林建宏
中文關鍵字 生物力學  紫杉醇  化療引發周邊神經病變  類線性黏彈理論  共軛聚焦顯微鏡光學切片術  三维影像模型重建  有限元素法 
英文關鍵字 Biomechanics  paclitaxel  CIPN  quasi-linear viscoelastic theory  finite element method  3D model reconstruction  AFM  confocal microscope 
學科別分類
中文摘要 癌症為一種細胞不正常再生並入侵或轉移至其他組織的疾病並且為全球第二大死因,在癌症的化學藥物治療中,常誘發周邊神經產生病變並迫使降低藥物劑量或終止療程而造成病人的死亡。然而目前此病變作用機轉仍未有定論,故本研究以力學之觀點探討此問題。
本研究以紫杉醇施加於PC-12類神經細胞以模擬人體內神經細胞受紫杉醇之影響,引入臨床現行施打計劃下血漿中的紫杉醇濃度,探討兩種施打計畫對活體PC-12細胞黏彈性力學的影響。以原子力顯微鏡對受損之細胞進行應力鬆弛試驗,並分別以類線性黏彈模型與有限元素法估測細胞黏彈之力學性質。另外,應用共軛對焦顯微鏡建立包含細胞本體與細胞核之三維有限元素模型,並以軸對稱模型所得材料參數模擬單顆細胞三維應力鬆弛。類線性黏彈模型分析顯示細胞結構阻尼參數短時注射組之變化大於長時注射組,表示短時注射對細胞之黏滯性影響較大。有限元素分析顯示藥物濃度改變對細胞核之機械性質之影響不明顯,藥物主要影響細胞胞漿,驗證紫杉醇改變細胞內微管分佈從而造成機械性質之改變。切楊氏模數結果發現短時注射組於施打過程中(12hr)有最劇烈之硬度變化並於施打結束後使細胞變軟,長時注射則是先使細胞變軟後於施打結束後趨於正常值。

關鍵字:生物力學、紫杉醇、化療引發周邊神經病變、類線性黏彈理論、共軛聚焦顯微鏡光學切片術、三維影像模型重建、有限元素法
英文摘要 Chemotherapy treatment of cancer by paclitaxel often results in peripheral neuropathy which is a main reason for patients to withdraw from therapy and reduce the survival rate. Most past researches of chemotherapy induced peripheral neuropathy (CIPN) focused on biochemical field. The goal of this thesis is using finite element (FE) method and quasi-linear viscoelasticity (QLV) theory to analyze the indentation of PC-12 cells subjected to in vitro simulations of two clinical paclitaxel doses. Mechanical properties of the cells were measured by curve fitting to both QLV theory and a FE model. Stress-strain relationship was derived from hyperelastic part for comparison purpose. Full 3D cell geometry model was built from confocal microscope immunofluorescences images of the cells. QLV results show that apparent Young’s modulus rises at 12 hours then goes down at 18 hours for 3-hr infusion groups and rises at 6 and 18 hour for 24-hr infusion groups. Viscous parameter C rises at 6, 12 hours and goes down for 3-hr group. FE results show that relaxation modulus at cytoplasm changes with the decreasing of paclitaxel concentration while that of the nucleus remains unchanged. From the stress-strain results, the Young’s moduli of the cytoplasm are lower than that of control groups for both 3-hr and 24-hr infusion groups, it implies that cytoplasm gets softer after treated by paclitaxel. At nucleus, the stiffness rises at 12 hours for the 3-hr infusion groups and rises from 6 to 18 hour for the 24-hr groups. In conclusion, both stiffness and viscous properties have larger changes in the 3-hr infusion group than the 24-hour group, suggesting that the cells were damaged more severely due to short time infusion dosing schedule. The soften of paclitaxel treated cytoplasm is consistent with the fact that paclitaxel would disrupt the microtubules of the cell. FE simulation results suggest that, regarding the peak indentation force, the axisymmetric model yield similar results as that of the full 3D model, saving the computation effort very much.
Key words: Biomechanics, paclitaxel, CIPN, quasi-linear viscoelastic theory, finite element method, 3D model reconstruction, AFM, confocal microscope
論文目次 摘要 i
誌謝 x
目錄 xi
圖目錄 xiii
表目錄 xvii
符號表 xix
第一章 緒論 1
1.1 化療引起的神經病變 1
1.2 類神經細胞與細胞機械特性 4
1.3 太平洋紫杉醇 5
1.4 細胞力學簡介 6
1.5 原子力顯微鏡掃描術 7
1.6 共軛聚焦顯微鏡光學切片術 8
1.7 有限元素分析 10
1.8 生物力學結合有限元素法研究 11
1.9 研究動機與目的 14
第二章 研究方法 15
2.1 PC-12類神經細胞培養及軸突誘發程序 17
2.1.1 PC-12類神經細胞培養 17
2.1.2 PC-12類神經細胞軸突誘發 19
2.2 紫杉醇濃度配製和施打計畫 21
2.3 原子力顯微鏡掃描實驗 23
2.3.1 原子力顯微鏡工作原理 23
2.3.2原子力顯微鏡硬體設備及操作 26
2.3.3 原子力顯微鏡力量感測系統 28
2.3.4 壓電掃描器潛變校正 29
2.4 PC-12類神經細胞機械性質量測 31
2.4.1 活細胞前處理 31
2.4.2 應力鬆弛試驗流程與壓印點選取 31
2.4.3 壓印試驗接觸點尋找 34
2.5 類線性黏彈模型擬合 39
2.6 共軛聚焦顯微鏡三維影像重建 44
2.6.1 細胞固定及免疫螢光染色 44
2.6.2 細胞本體與細胞核面積、形貌與位置及三維影像重建 46
2.7 有限元素法逆向分析 47
2.7.1 細胞影像與二維幾何模型建立 47
2.7.2 網格與元素 49
2.7.3 材料本構方程式與參數給定 52
2.7.4 細胞介面性質 54
2.7.5 分析部負載與邊界條件 55
2.7.6 逆向有限元素最佳化參數擬合 55
第三章 結果 58
3.1 活細胞前處理 58
3.2 壓電致動器潛變修正 60
3.3 類線性黏彈理論參數擬合 64
3.4 逆向有限元素參數分析 75
3.5三維細胞模型重建及有限元素模擬 99
3.5.1 共軛焦細胞顯微影像前處理結果 99
3.5.2 三維細胞立體模型建立 104
3.5.3 三維有限元素模型模擬結果 106
第四章 討論 108
4.1 類線性黏彈模型與有限元素模型比較 108
4.2 藥物濃度之探討 110
4.3 針尖半徑對類線性黏彈模型擬合之影響 111
4.4 針尖半徑對有限元素模型擬合之影響 114
4.5光學影像與AFM細胞高度差異 119
4.6 有限元素模型之缺點 120
第五章 結論與建議 122
5.1結論 122
5.2 建議 123
附錄 A 逆有限元素法最佳化數據表 124
參考文獻 127
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