臺灣博碩士論文加值系統

本研究以界達電位粒徑分析儀(zetasizer 3000)與高效毛細管電泳儀(high-performance capillary electrophoresis, HPCE)探討以可可脂(cocoa butter, CB)和棕櫚脂(tripalmitin)為脂質相的固體脂質奈米粒子(solid lipid nanoparticle, SLN)、以L-arginine對SLN表面改質形成的R-SLN和人腦微血管內皮細胞(human brain microvascular endothelial cells, HBMECs)在電磁場照射下的電泳遷移率與柔軟參數變化，利用TEM與FE-SEM觀察粒子外觀並以小角度X光散射儀(SAXS)得知SLN表面聚電解質層厚度，進一步結合柔軟粒子電泳理論計算SLN、R-SLN和HBMECs的固定電荷密度(fixed charge density)與柔軟參數(softness)。經由電子顯微鏡下觀察到SLN與R-SLN的型態為圓滑球型而且粒子大小均一，SLN脂質相中可可脂含量增加，造成SLN粒徑變大、電泳遷移率絕對值增加與柔軟參數下降。R-SLN表面上越多Arg造成電泳遷移率絕對值下降、增加聚電解質層厚度、柔軟參數增加與提高被HBMECs攝入的能力。電磁場效應造成SLN、R-SLN和HBMECs的電泳遷移率絕對值下降與柔軟參數上升。

The study aims to the electrophoretic mobility and the softness of solid lipid nanoparticles (SLN) composed of cocoa butter and tripalmitin as lipid phase, L-arginine loaded SLN (R-SLN), and human brain microvascular endothelial cells (HBMECs) were investigated under the influences of electromagnetic field with various power. The size of SLN and R-SLN were measured by zetasizer and confirmed by the images of FE-SEM and TEM. The softness, fixed charge density, and Donnan potential were estimated from the electrophoretic mobility of capillary electrophoresis and the thickness of polyelectrolyte layer of small angle X-ray scattering (SAXS) with the soft particle electrokinetic theory. The results revealed that, the size and the electrophoretic mobility of SLN increased with an increase in the content of CB, but the softness of SLN decreased. The electrophoretic mobility of R-SLN reduced with an increase in L-arginine adsorbed onto SLN surface, but the softness of R-SLN raised. For EMF effects, the electrophoretic mobility of SLN, R-SLN, and HBMECs decreased, but the softness increased.

中文摘要 I
英文摘要 II
目錄 III
圖目錄 VI
符號說明 VIII
第一章 緒論 1
1.1 細胞、膠體電動力學分析 1
1.2 研究動機與目的 2
第二章 文獻回顧 3
2.1 電動力學 3
2.1.1 電雙層與界達電位 3
2.1.2 毛細管電泳發展與原理 4
2.1.3 毛細管電泳應用於膠體分析 5
2.1.4 毛細管電泳應用於細胞分析 6
2.1.5 電泳遷移率計算 8
2.1.6 固定電荷密度與Donnan電位計算 8
2.1.7 柔軟粒子理論分析 9
2.2 固體脂質奈米粒子(Solid lipid nanoparticle, SLN) 11
2.3 精胺酸 14
2.4 電磁場於膠體溶液與生物醫學的應用 15
第三章實驗材料、儀器及步驟 19
3.1 實驗材料 19
3.1.1 SLN製備材料 19
3.1.2粒徑、zeta potential與電泳遷移率測定 20
3.1.3 TEM樣品處理材料 20
3.1.4人腦微血管內皮細胞培養與冷凍保存 21
3.1.5免疫螢光法 23
3.1.6其他實驗器具和耗材 23
3.2 實驗儀器 25
3.3 實驗步驟 29
3.3.1 SLN製備 29
3.3.2 Tris緩衝液製備 30
3.3.3粒子密度與表面覆輔v計算 30
3.3.4 R-SLN的製備 31
3.3.5 SLN和R-SLN的粒徑、界達電位 32
3.3.6 SLN和R-SLN電泳遷移率的測定 33
3.3.7穿透式電子顯微鏡 (Transmission electron microscope, TEM) 33
3.3.8場發射型掃描式電子顯微鏡 (Field emission scanning electron
microscopes, FE-SEM) 34
3.3.9小角度X-Ray繞射儀 (Small Angle X-Ray Scattering, SAXS) 34
3.3.10鼠尾膠原蛋白溶液的製備與舖盤 35
3.3.11細胞培養基配製 35
3.3.12人腦微血管內皮細胞培養 36
3.3.13 HBMECs於電磁場下攝入R-SLN，測定界達電位、電泳遷移率 37
3.3.14螢光分析HBMECs攝入SLN、R-SLN 37
第四章 結果與討論 39
4.1改變脂質相中可可脂比例對SLN的影響 39
4.2 改變L-arginine覆輔v對R-SLN的影響 41
4.3 電磁場對SLN、R-SLN的影響 43
4.4 電磁場對人腦微血管內皮細胞與SLN、R-SLN共培養時的影響 44
第五章 結論與建議 47
5.1 結論 47
5.2 建議 48
參考文獻 49
附錄A：柔軟粒子電泳理論的參數 78
附錄B：精胺酸對UV吸收度的檢量線 79
附錄C：SLN成份的結構式 80
附錄D：精胺酸於不同pH值溶液中結構式的改變 82

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