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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Title:	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
Authors:	魏延麟;Wei,Yen-Lin
Contributors:	生物醫學工程研究所
Keywords:	超聲波駐波場;聚苯乙烯微球;自體組裝;逐層堆疊;分子傳遞;Ultrasonic standing wave fields;Polystyrene microsphere;Self-assembly;Layer by layer;Molecular transport
Date:	2013-08-28
Issue Date:	2013-10-08 15:10:31 (UTC+8)
Publisher:	國立中央大學
Abstract:	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。 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.
Appears in Collections:	[Institute of Biomedical Engineering] Electronic Thesis & Dissertation

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