臺灣博碩士論文加值系統

將棕櫚酰殼聚醣(NPCS)、超順磁氧化鐵奈米顆粒(SPIO)、抗癌藥物阿黴素
(DOX) 利用電噴霧技術製備一種新型的藥物載體微米顆粒。NPCS 的特性透過核
磁共振光譜(NMR)以及膨潤性測試所製備的殼聚醣改質物具有pH 標靶的特性。
DOX-SPIO/NPCS 微米顆粒的顆粒型態以及粒徑尺寸利用穿透式以及掃描式電子
顯微鏡進行分析。結果顯示在電噴霧中使用甲酸/丙酮複合溶劑在顆粒的成形效
果相較於純甲酸溶劑較好。所製備的微米顆粒的粒徑尺寸為185 ± 87 nm。之後
利用傅立葉紅外線光譜(FT-IR)鑑定DOX-SPIO/NPCS 顆粒材料表面性質，所製備
的微米顆粒經過電噴霧技術不會改變材料的性質。在穿透式電子顯微鏡(TEM)
以及核磁共振T2影像(MRI)分析結果其超順磁氧化鐵奈米顆粒(SPIO)成功被包覆
於微米顆粒載體中。在體外藥物釋放實驗中，DOX-SPIO/NPCS 藥物載體顆粒相
較於鹼性環境中酸性環境(pH 6.5)會加速藥物的釋放量。微米顆粒在細胞毒性分
析(MTT 測試)中顯示DOX-SPIO/NPCS 微米顆粒相較於藥物DOX具有更好的抗
腫瘤能力，除此之外，當DOX-SPIO/NPCS 微米顆粒包覆5μg/mL DOX 藥物在
酸性環境(pH 6.5)中相較於鹼性環境中能更有效殺死HepG2 肝癌細胞。綜合以上
解結果所製備的DOX-SPIO/NPCS 微米顆粒具有相當的潛力作為pH 應答型藥物
傳輸系統應用於肝癌的治療。

A novel pH-sensitive drug delivery micro-particles, based on N-palmitoyl
chitosan (NPCS) using electrospray technology to transport the superparamagnetic iron oxide (SPIO) and anticancer drug doxorubicin (DOX) was developed. The characteristic of NPCS was characterized by nuclear magnetic resonance(NMR) and swelling testing, it showed that modified chitosan had pH-targeted property. The morphology and size of the DOX-SPIO/NPCS micro-particles were investigated by transmission and scanning electron microscopy. Based on using different vaporization properties of electrospray solvent showed formic acid/acetone co-solvent has better
particle forming ability than formic acid solvent alone in suspension collection. The micro-particles had the diameters in 185 ± 87 nm. Materials surface properties of DOX-SPIO/NPCS micro-particles, was confirmed using Fourier Transform Infrared Spectroscopy (FTIR) to make sure that using electrospray technology will not change the materials properties. Transmission electron microscopy (TEM) and nuclear magnetic resonance T2 imaging(NMRI) confirmed the SPIO in composite micro-particles. In drug release profile, we found the DOX-SPIO/NPCS drug delivery micro-particles was accelerated in acidic environment (pH 6.5). Micro-particles in
cytotoxicity assay (MTT assay) showed that DOX-SPIO/NPCS micro-particles has
better anti-tumor ability than that of DOX freeform. Additionally, micro-particles were loaded 5 μg/mL DOX in acidic environment (pH 6.5) to kill HepG2 cell effectively. The DOX-SPIO/NPCS micro-particles could be potentially applied as pH sensitive drug delivery system for Hepatic carcinoma therapy.

中文摘要.................................................................. I
英文摘要................................................................. II
全名與縮寫對照表 ......................................................... IV
致謝...................................................................... V
目次..................................................................... VI
圖次..................................................................... IX
表次.................................................................... XII
第1 章 緒論............................................................... 1
1-1. 前言................................................................. 1
1-2. 實驗動機與目的........................................................ 1
1-3. 實驗流程與架構........................................................ 2
第2 章 文獻回顧............................................................ 4
2-1. 肝癌................................................................. 4
2-1.1 肝癌................................................................ 4
2-1.2 肝癌療法與抗癌藥物.................................................... 5
2-1.3 阿黴素(Doxorubicin, DOX) ........................................... 8
2-2. 奈米科技的應用與製備技術............................................... 9
2-2.1 奈米藥物於醫學上的應用................................................ 9
2-2.2 電噴霧製備技術...................................................... 11
2-3. 生物醫學材料簡介...................................................... 13
2-3.1 殼聚醣與棕櫚酰殼聚醣(N-palmitoyl Chitosan, NPCS) .................... 13
2-3.2 超順磁氧化鐵(Super Paramagnetic Iron Oxide, SPIO) .................. 14
2-3.3 pH 應答型材料於癌症環境 ............................................. 15
第3 章 材料與研究方法 ..................................................... 20
3-1. 實驗藥品............................................................. 20
3-2. 實驗儀器............................................................. 22
3-3. 電噴霧技術製備DOX-SPIO/NPCS 顆粒 ..................................... 24
3-4.1 製備棕櫚酰殼聚醣(N-palmitoyl Chitosan, NPCS) ........................ 24
3-4.2 製備DOX-SPIO/NPC 藥物載體顆粒 ....................................... 24
3-4. NPCS 物理性質分析.................................................... 25
3.4.1 核磁共振光譜(Nuclear Magnetic Resonance, NMR) ...................... 25
3.4.2 不同酸鹼條件下NPCS 膨潤性質.......................................... 25
3-5. 電噴霧DOX-SPIO/NPCS 顆粒材料分析 ..................................... 26
3-5.1 利用傅立葉紅外光譜(Fourier-Transform Infrared Spectrometer,
FT-IR)化學結構分析........................................................ 26
3-5.2 電子顯微鏡(Field-Emission Scanning Electron Microscope, FE-SEM)
於DOX-SPIO/NPCS 顆粒型態分析 ............................................. 26
3-5.3 使用穿透式電子顯微鏡(Transmission electron microscope, TEM)進
行DOX-SPIO/NPCS 顆粒尺寸分析 ............................................. 27
3-5.4 核磁共振T2 影像成像實驗(Nuclear Magnetic Resonance Imaging,
NMRI) ...................................................................27
3-6. 阿黴素標準曲線建立................................................... 28
3-7. 阿黴素包覆率計算........... 28
3-8. DOX-SPIO/NPCS 顆粒之體外釋放曲線 .................................... 29
3-9. 肝癌細胞株HEPG2 .................................................... 30
3-9.1 細胞種類........................................................... 30
3-9.2 細胞培養基配置...................................................... 30
3-9.3 細胞培養........................................................... 31
3-9.4 冷凍細胞........................................................... 32
3-9.5 活化細胞........................................................... 32
3.9.6 細胞計數........................................................... 32
3-10. 體外抗腫瘤細胞療效試驗評估........................................... 33
3-10.1 不同藥物濃度對於腫瘤細胞之影響...................................... 34
3-11 實驗統計分析........................................................ 34
第4 章 結果與討論 ....................................................... 35
4-1. NPCS 物理性質分析 .................................................. 35
4-1.1 核磁共振光譜分析.................................................... 35
4-1.2 不同pH 值下NPCS 的體積變化 ......................................... 36
4-2. DOX-SPIO/NPCS 顆粒材料性質分析 ...................................... 38
4-2.1 FT-IR 分析結果與討論 ............................................... 38
4-2.2 SEM 顆粒型態分析 ................................................... 39
4-2.3 TEM 分析結果與討論 ................................................. 41
4-2.4 MRI 實驗結果與討論 ................................................. 41
4-3. 阿黴素藥物包覆率計算.................................................. 43
4-4. DOX-SPIO/NPCS 顆粒體外釋放曲線測試與分析 ............................. 44
4-5. 體外抗腫瘤細胞療效試驗評估............................................ 46
第5 章 結論.............................................................. 48
第6 章 未來展望.......................................................... 50
參考文獻................................................................. 51

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