近年來生醫材料已廣泛應用於標靶治療之細胞、組織工程及生醫感測等新興醫療器材和生物技術產業上，亦屬政府推動重點產業之要項，而藥物釋放技術在近年來已廣泛應用於生醫材料領域上，為最近幾年發展迅速之給藥型態。
　　癌症數國人十大死因榜首，對於許多癌症以前是採單一種藥物治療的方式，結果病人服藥量多、副作用大，療效卻有限。現在治療癌症不再是使用單一化療藥物，而是聯合多種藥物﹑以不同機制進行治療，此治癌新策略也稱作雞尾酒療法。
　　在此我們選用大腸癌細胞(HCT-8)作為研究目標，提出了一個整合性的生醫晶片，我們發展了一合併試驗平台，該平台允許高通量試驗、但藥物劑量低，減少昂貴的化療藥物使用量。並且我們可以個人化的進行雞尾酒藥物篩選，找到最適合患者的藥物比例。
　　在本實驗中我們使用水膠 PEGDA (poly(ethylene glycol)diacrylate)，為PEG改質成兩端具有烯基雙鍵(acrylate or methacrylate) 因此可經由照射UV光交聯。利用水膠照光會固化的特性加上光罩部分曝光，可以形成包藥水膠液珠陣列，達到定量的目的。而最後定量好的包藥水交會直接對準合併細胞培養晶片，使藥物同時平行釋放到細胞中。此晶片操作快速，準確性高，而且少量之樣品即足以供分析之用，測定範圍廣。我們可以合併五種藥物在動態範圍千倍之下，藥物完整釋放僅需30秒，完成高通量的試驗。我們同時以二維以及三維方式培養細胞，同時比較IC50可以看出以三維培養的細胞抗藥性較高，可以歸咎於二為培養非接近體內自然狀態生長，三維培養較接近真實腫瘤狀態。最後雞尾酒結果顯示出我們的晶片是接近傳統手動配藥結果，證實晶片可行性。

In recent years, biomedical materials have been widely applied in the treatment of the target cells. While tissue engineering and biomedical sensing and other new medical devices and biotechnology industries also belongs to the Government’s promotional key point. And drug delivery technology in recent years has been widely developed in the medical material field. Cell-based assays have been an important pillar of the drug discovery process to provide a simple, fast, and cost-effective tool to avoid large-scale and cost-intensive animal testing.
In our study we introduces a combinatory assay platform that allows high-throughput but low-drug-dosage screening of five anti-cancer drugs as a cocktail for personalized cancer treatment. Photosensitive PEGDA hydrogel was employed for drug dosage definition through drop array formation and selective UV crosslinking process. The finally defined cocktail drugs in hydrogel will be directly released in parallel when combined with cell chips. This device is capable to combine 5 drugs with 1000 folds dynamic range in 30 second with low drug consumption for in-parallel cocktail screening process.
And it is imperative to establish in vitro cell-based systems that can more realistically mimic the in vivo cell behaviors and provide more predictable results to in vivo tests. We found that cells in the 3D culture environment differ morphologically and physiologically from cells in the 2D culture environment. Drugs were highly active in 2D monolayer culture but less active and gradually lost their activity in 3D spheroids. Our cocktail chip results are analogous to a conventional method.

總目錄
第一章 緒論 1
1.1 研究背景 1
1.2 研究動機 2
1.3 研究目的 2
第二章 文獻回顧 4
2.1雞尾酒藥物療法 4
2.2 大腸癌簡介 4
2.2.1 大腸癌藥物治療 6
（一）化學藥物 6
（二）標靶藥物 8
2.3水膠簡介 9
2.3.2水膠的應用 13
2.3.3水膠定量以及擴散釋放 16
2.4液珠成型方式 18
2.5藥物混合方法 20
2.5.1連續式混合 20
2.5.2平行式混和藥物 22
第三章 晶片設計 25
3.1 晶片設計 25
3.2 液珠陣列成型 26
3.3 演算法介紹 27
第四章 實驗步驟、設備與材料 29
4.1親疏水性晶片製程流程 29
4.1.1 詳細流程 29
4.2水膠原料與配製 32
4.2.1水膠與光起始劑配製溶液 32
4.2.2水膠固化 33
4.3 細胞培養 34
4.3.1 附著型細胞(adherent cell) 34
4.3.2 懸浮型細胞(suspension cell) 35
4.3.3 融合瘤﹙hybridoma﹚ 35
4.3.3 三維細胞培養(3D cell culture) 36
4.4紫外線/可見光分光光譜儀分析定量 37
4.5半數抑制濃度(IC50)計算 38
4.6細胞計數 38
第五章 實驗結果與討論 40
5.1液珠成型測試 40
5.2 PEGDA毒性測試 41
5.3水膠包覆螢光擴散 43
5.3.1.螢光偵測法 43
5.3.2 Rhodamine 6G 溶液的配製 43
5.3.3水膠濃度、照光時間不同的影響 45
5.3.4比較不同PEGDA分子量的影響 46
5.3.5可見光/紫外光分光光譜儀偵測法 47
5.3.6水膠包覆R6G擴散測試 48
5.4水膠包覆五種藥物釋放 48
5.5 IC50測量 50
5.6雞尾酒測試 54　
第六章 結論、未來工作 57
6.1 總結 57
參考文獻 58
 

[1] Lapidot, T et.al,”A cell initiating human acute myeloid leukaemia after transplantation into SCID mice.”,Nature, 1994
[2] Young-Hyun Jin et.al, Lab-on-a-chip, 2009
[3] 社團法人中華民國愛滋感染者權益促進會, website
[4] Chih-Ming Ho, “Use of Fractional Factorial Designs in Antiviral Drug Studies.”, Wiley Online Library, 2012
[5] 中山大學附屬醫院腸胃科, website
[6] Qiu Y. , Park K. ,”Environment-sensitive hydrogels for drug delivery.”,Advanced Drug Delivery Reviews, 53, 321-339 ,2001
[7] S. Bushetti, V. Singh, S. Raju, Atharjaved, Veermaram,”Stimuli sensitive hydrogels: A review.”, Indian Journal of Pharmaceutical Education and Research. 43 3 241 250 0019-5464, 2009
[8] Brandon V. Slaughter, Shahana S. Khurshid, Omar Z. Fisher,
[9] Ali Khademhosseini, and Nicholas A. Peppas, “Hydrogels in Regenerative Medicine.”, Adv. Mater. 21, 3307–3329,2009
[10] Yuanting Xu, Li Li, Xixun Yu, Zhipeng Gu, Xu Zhang., “Feasibility study of a novel crosslinking reagent (alginate dialdehyde) for biological tissue fixation.” ,Carbohydrate Polymers, Vol.87 (2), pp.1589-1595, 2011
[11] Liu, V.A. and S.N. Bhatia, “Three-Dimensional Photopatterning of Hydrogels Containing Living Cells.”,Biomedical Microdevices 4(4): p.257-266., 2002.
[12] Jos Wennink, “Biodegradable Hydrogels by Physical and Enzymatic crosslinking of biomacromolecules.”, Lab on a Chip,1984
[13] Lan Wei, Chunhua Cai, Jiaping Lin, and Tao Chen, “Dual-drug delivery system based on hydrogel/micelle composites .”,Biomaterials ,30: 2606, 2009
[14] James M. Anderson, “Journal of Biomedical Materials Research Part B: Applied Biomaterials.”, Wiley Online Library,2012
[15] A.R.Parker and C.R.Lawrence,”Water Capture by a Desert Beetle.”,Nature,414,33-34,2001
[16] Pavel A. Levkin, “DropletMicroarray: facile formation of arrays of microdroplets and hydrogel micropads for cell screening applications.”, Lab on a Chip, 2012
[17] George M. Whitesides, “Fabricating Large Arrays of Microwells with Arbitrary Dimensions and Filling Them Using Discontinuous Dewetting.”,Anal. Chem. 1998, 70, 2280-2287,1999
[18] Young-Hyun Jin, “Lab-on-a-chip for multiplexed biosensing of residual antibiotics in milk.”, Lab on a Chip, 2009
[19] Shaojiang Zeng, “Microvalve-actuated precise control of individual droplets in microfluidic devices.”, Lab on a Chip, 2008
[20] Kang Yuejun, “Microfluidic synthesis of monodisperse PEGDA microbeads for sustained release of 5-fluorouracil.”, Lab on a Chip, 2014
[21] Su Eun Chung, “One-step pipetting and assembly of ECM for high-throughput multiplexed bioassays.”, Nature, 2014
[22] Martina Hitzbleck, “Capillary soft valves for microfluidics.”,Lab on a chip,2012
[23] JR Sanchez-Valencia, “Excitation transfer mechanism along the visible to the Near-IR inrhodamine J-heteroaggregates.”,Advance Article on the web ,2010
[24] Material Safety Data Sheet. Rhodamine 6G MSDS, sciencelab website