博碩士論文 993211012 詳細資訊




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姓名 魏延麟(Yen-Lin Wei)  查詢紙本館藏   畢業系所 生物醫學工程研究所
論文名稱 結合超聲波駐波場與層堆疊自體組裝微球載體建構提高分子傳遞至細胞內效率之方法
(Using Ultrasonic Standing Wave Fields in Association with Self-Assembly Microspheres to Enhance the Efficiency of Molecular Transport in vitro)
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摘要(中) 為了提高細胞治療成效同時能將所應用的藥物輸送系統放大規模,本研究嘗試結合自體組裝之聚苯乙烯微球載體與超聲波駐波場以建構出促進分子傳遞至細胞內效率之方法。本研究係利用硒化鎘 / 硫化鋅量子點模擬藥物分子,將其以層堆疊自體組裝的方式塗佈於聚苯乙烯微球載體之表面製成量子點微米球,並以顯微技術與螢光光譜儀分析成品。實驗結果證明 1) 量子點晶體能夠以 1.0 pmole/cm^2 的容量密度均勻散佈於載體表面, 2) 量子點微米球與獨立量子點也擁有近似的光學性質, 3) 該量子點與載體間的靜電作用可承受環境剪應力之干擾。接著,在確認細胞與微球載體能夠被駐波場牽引至聲壓節面後,我們訂定出最佳的超聲波照射時間為五分鐘,並且證明超聲波駐波場不會對細胞與微球載體造成損害。最後利用流式細胞儀分析細胞經或未經駐波場照射後量子點微球傳遞至其內部之效果,結果顯示出超聲波駐波場可提高含有量子點的細胞數量約 1.2 倍 (P < 0.01) 以及細胞螢光強度約 1.3 倍 (P < 0.01) 。本研究證明了結合超聲波駐波場與微球載體有助於提升載體表面上的分子進入細胞內之效率。
摘要(英) To enhance the cellular therapy efficacy and scale up the drug delivery system used, we aimed to develop a complex molecular delivery system comprising ultrasound standing wave fields (USWF) and microsphere techniques. In this study, CdSe/ZnS quantum dots (QDs) were used to imitate drug molecules and the QDs-coated polystyrene microspheres were prepared through layer-by-layer approach. The developed QDs-coated microspheres were characterized using microscopy and spectrofluorometry, and exhibited that 1) QDs can entirely cover the surface of microspheres with uniform distribution in a coverage rate of 1.0 pmole/cm^2, 2) QDs-covered microspheres exhibited similar optical properties with isolated QDs, and 3) the electrostatic interactions between QDs and microsphere surfaces were robust enough to resist mechanical stress induced by ultrasound. After determining the optimal USWF exposure time of 5 minutes in which the cellular viability was > 90% within 48 h, we examined the efficiency of microspheres internalization of the DH82 macrophages with and without USWF treatment using flow cytometry. Our results showed that the cells with USWF exhibited 1.2-fold (P < 0.01) and 1.3-fold (P < 0.01) higher than the group without USWF in terms of fluorescence-expressed cell number and fluorescence intensity from the cells, respectively. The system of USWF in association with microspheres developed in this study provided a feasible means for enhancement of molecular transport efficiency in vitro.
關鍵字(中) ★ 超聲波駐波場
★ 聚苯乙烯微球
★ 自體組裝
★ 逐層堆疊
★ 分子傳遞
關鍵字(英) ★ Ultrasonic standing wave fields
★ Polystyrene microsphere
★ Self-assembly
★ Layer by layer
★ Molecular transport
論文目次 摘要 i
Abstract ii
目錄 iii
圖目錄 vi
表目錄 ix
第一章 緒論 1
第二章 文獻回顧 2
2.1 量子點塗佈微球載體 2
2.1.1 量子點簡介 3
2.1.2 量子點在細胞生物學之應用 8
2.1.3 層堆疊自體組裝合成策略 15
2.2 超聲波駐波場 19
2.2.1 超聲波與壓電效應 20
2.2.2 超聲波在醫學領域之應用 22
2.2.3 超聲波駐波場簡介 25
第三章 實驗藥品、儀器設備與研究方法 28
3.1 實驗藥品 28
3.2 儀器設備 29
3.3 細胞培養 31
3.4 超聲波裝置 32
3.5 研究方法 34
3.5.1 合成親水性 CdSe/ZnS 量子點 34
3.5.2 製作 CdSe/ZnS 量子點塗佈微球載體 35
3.5.3 CdSe/ZnS 量子點塗佈微球載體的光學測定 37
3.5.4 CdSe/ZnS 量子點塗佈微球載體的形態測定 38
3.5.5 CdSe/ZnS 量子點塗佈微球載體的穩定性測定 39
3.5.6 超聲波駐波場影響細胞存活率之測定 41
3.5.7 超聲波駐波場促進微球載體表面分子傳遞至細胞內之測定 43
3.5.8 統計分析 45
第四章 結果與討論 46
4.1 CdSe/ZnS 量子點塗佈微球載體於製程中的電位分析 46
4.2 CdSe/ZnS 量子點塗佈微球載體的形態與光學性質分析 49
4.3 CdSe/ZnS 量子點塗佈微球載體的表面容量分析 51
4.4 CdSe/ZnS 量子點塗佈微球載體的穩定性分析 54
4.5 細胞與 PS 微球載體被超聲波駐波場牽引至節面之現象 56
4.6 細胞經超聲波駐波場曝照後的存活率分析 60
4.7 超聲波駐波場提高載體表面分子傳遞至細胞內效率之分析 62
第五章 結論 66
第六章 參考文獻 67
附錄 75
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指導教授 李宇翔(Yu-Hsiang Lee) 審核日期 2013-8-28
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