臺灣博碩士論文加值系統

本研究是利用環境敏感型高分子製備奈米顆粒並應用於藥物釋放。系統架構是利用酸鹼敏感型天然高分子-幾丁聚醣，在醋酸水溶液中進行自組裝，形成幾丁聚醣/醋酸錯合微胞。接續在微胞中加入氮-異丙基丙烯醯胺單體，以過硫酸鉀在高溫下起始反應，由於生成的聚(氮-異丙基丙烯醯胺)為溫度敏感型高分子，當反應溫度高於其最低臨界溶解溫度時，高分子鏈產生疏水性而縮進微胞中，最後交聯外層幾丁聚醣，完成[幾丁聚醣/醋酸]-聚(氮-異丙基丙烯醯胺)奈米顆粒製備。
實驗的第一部分討論幾丁聚醣/醋酸微胞的成形條件、形態觀察與粒徑分析。首先使用螢光光譜儀並以芘做為疏水探針，測量幾丁聚醣/醋酸之臨界微胞濃度，討論醋酸濃度與氮-異丙基丙烯醯胺單體，對於臨界微胞濃度的影響。接續使用穿透式電子顯微鏡與動態粒徑分析儀研究微胞的形態與粒徑。
實驗的第二部分為[幾丁聚醣/醋酸]-聚(氮-異丙基丙烯醯胺)奈米顆粒的製程研究。藉由調控單體進料量、交聯劑進料量、交聯溫度、反應溫度等參數討論對於顆粒形態、粒徑、界面電位的影響。顆粒溫感性質的測試是使用紫外光-可見光分光光譜儀，量測該顆粒之最低臨界溶解溫度；並利用動態粒徑分析儀，計算顆粒在升溫前後粒徑的收縮比率，並測試是否具有溫感可逆性。
實驗最後的部分為[幾丁聚醣/醋酸]-聚(氮-異丙基丙烯醯胺)奈米顆粒在藥物釋放的應用。模擬藥物選用鹽酸四環素，控制載藥環境進行包覆96小時，接續以離心的方式分離含藥顆粒，並透過上清液的未包覆藥物量推算藥物於顆粒含量及藥物包覆效率，同時討論初始載藥量對於包覆的影響。將含藥顆粒分散在不同環境(酸鹼值、溫度、鹽類濃度、酵素、自由基)的生理食鹽水中，探討各個變因對於累積釋放比率的影響，藉以了解顆粒對於藥物控制釋放的能力。

In this research, the stimuli-responsive polymers: chitosan(CS) and poly(N-isopropylacrylamide)(PNIPAAm) were used to synthesize the nanoparticles as a drug carrier. The amount of pH-sensitive polymer, chitosan, was adjusted to a proper molar ratio with acetic acid(HAc) to form CS/HAc micelles by self-assembly. Next, NIPAAm was introduced into the system and initialized by potassium persulfate(KPS) at high temperature which was higher than Lower Critical Concentration Temperature(LCST) of PNIPAAm; therefore, PNIPAAm polymer chain would become hydrophobic and shrink into micelles. The [CS/HAc]-PNIPAAm particles were eventually completed by cross-linking the the out-layer chitosan.

The first part of this research was focused on the formation conditions of CS/HAc micelles, including the measurements of Critical Micellization Concentration(CMC), morphologies, and size distribution. CMC was measured by fluorescence spectrometer and pyrene was used as a hydrophobic probe. The effect of the concentration of acetic acid and presence of NIPAAm were studied. The observation of morphologies and size distribution were obtained with transmission electron microscopy(TEM) and dynamic light scattering(DLS), respectively.

The second part was the synthesis of [CS/HAc]-PNIPAAm nanoparticle. The morphology, particle size distribution, and zeta-potential of the nanoparticles were discussed by manipulate several parameters such as concentration of NIPAAm and cross-linking agent, cross-linking temperature, and polymerization temperature. The thermo-sensitivity and LCST of particles were tested by UV-Vis spectrophotometer. Dynamic light scattering technique was also applied to examine the particle size-reversible ability.

Finally, Doxycycline Hyclate was chosen as stimuli-drug and the drug-loaded particles were prepared under stable environmental conditions. After loading drug for 96hrs, the drug-loaded particles and the rest of stimuli-drug solution were separated via centrifugation process. The drug-loading content and the encapsulation efficiency were calculated via measuring the drug content in the upper phase of centrifugate. Furthermore, effect of initial drug-loading concentration on the encapsulation efficiency was discussed. Drug-loaded particles were dispersed in normal saline which was controlled in different conditions such as temperature, pH, concentration of salt, and the presence of enzyme and free radical. The release properties of drug-loaded particles in aforementioned conditions were studied.

論文口試委員會審定書 i
致謝 ii
中文摘要 I
Abstract II
目錄 IV
圖目錄 VII
表目錄 X
第一章 緒論 1
1-1前言 1
1-2研究目的 1
第二章 文獻回顧 2
2-1環境敏感型高分子材料簡介與應用 2
2-1-1環境敏感型高分子材料定義與分類 2
2-1-2溫度敏感型高分子材料簡介 3
2-1-3酸鹼(pH)敏感型高分子材料簡介 9
2-1-4其他環境敏感型高分子材料簡介 13
2-2 奈米顆粒合成技術與未來發展趨勢 15
2-2-1 奈米顆粒合成技術之演進 15
2-2-2 奈米顆粒之未來發展趨勢 30
2-3 藥物傳輸載體與控制釋放系統簡介 33
2-3-1藥物傳輸系統與藥物控制釋放原理 33
2-3-2藥物控制釋放傳輸系統 35
第三章 實驗方法 43
3-1實驗藥品 43
3-2實驗儀器 47
3-3實驗步驟 49
3-3-1實驗流程圖 49
3-3-2幾丁聚醣/醋酸微胞 50
3-3-2-1臨界微胞濃度測試 50
3-3-2-2幾丁聚醣/醋酸微胞粒徑與表面電位分析 51
3-3-2-3幾丁聚醣/醋酸微胞形態觀察 52
3-3-3 [幾丁聚醣/醋酸]-聚(氮-異丙基丙烯醯胺)奈米顆粒 53
3-3-3-1合成方法 53
3-3-3-2最低臨界溶解溫度測試及轉化率分析 54
3-3-3-3奈米顆粒粒徑與表面電位分析 55
3-3-3-4奈米顆粒形態觀察 55
3-3-4含藥奈米顆粒製備及藥物釋放實驗 56
第四章 結果與討論 58
4-1幾丁聚醣/醋酸微胞 58
4-1-1微胞成形機制 58
4-1-2臨界微胞濃度分析 58
4-1-2微胞形態觀察及粒徑分析 59
4-2 [幾丁聚醣/醋酸]-聚(氮-異丙基丙烯醯胺)奈米顆粒 60
4-2-1顆粒合成機制 60
4-2-2氮-異丙基丙烯醯胺進料量的影響 60
4-2-3交聯劑的影響 62
4-2-4聚合溫度的影響 64
4-2-5奈米顆粒溫感性質測試 64
4-3含藥奈米顆粒製備及藥物釋放實驗 65
4-3-1含藥奈米顆粒製備 65
4-3-2藥物釋放實驗 65
4-3-2-1 溫度與酸鹼值對奈米顆粒藥物釋放的影響 65
4-3-2-2 藥物進料量對奈米顆粒藥物釋放的影響 67
4-3-2-3 鹽類濃度對奈米顆粒藥物釋放的影響 67
4-3-2-4 分解幾丁聚醣對奈米顆粒藥物釋放的影響 68
第五章 結論 88
參考文獻 90

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