臺灣博碩士論文加值系統

高度惡性神經膠質瘤是最常見侵略性高的原發性腦腫瘤，根據統計全球平均十萬人中有3到5名患者，即便接受各種治療，患者癒後存活率也無太大的改善，現今治療腦種瘤方式以外科手術切除為主，然而其神經膠質瘤的向外浸潤的特性，造成術後極高的復發風險。因此術後多會以輔助性化療抑制腦中殘留的腫瘤。然而經由口服或注射進入人體的藥物遇到最大的問題在於腦中有血腦屏障 ( blood brain barrier, BBB ) 的結構大幅限制藥物由血液循環滲透到腦組織，經過循環代謝難以維持以有效的藥物濃度，因此發展有別於傳統的給藥方式。近年來，利用生物可分解的生醫材料攜帶藥物，直接於手術後進行顱內給藥是重要的研究方向，如臨床上的 Gliadel® Wafer，該藥物載體將直接在腦中進行藥物控制釋放，使目標區域周邊能維持有效藥物濃度並長期存在，增加藥物的治療效果。
本研究，使用氧化透明質酸水膠（Oxidized hyaluronic acid-based hydrogel）做親水性藥物卡鉑（Carboplatin）的藥物載體結合放射線治療神經膠質瘤，探討釋放載卡鉑藥物過程中結合放射線治療神經膠質瘤，改善臨床產品遇到的問題提供新的選擇治療方式，提高存活率。
研究結果顯示，經過改質後的氧化透明質酸與己二酸二醯肼形成水膠，並從( fourier transform infrared spectroscopy, FTIR ) 光譜圖分析其官能基的變化，從 ( electronic data systems, EDS ) 元素分析中也可以得知藥物是存在於水膠當中，在流變儀分析中利用溫度的控制延長其成膠時間，在藥物的釋放中與治療的時間規劃符合，經由 IC50 確認給予藥物的濃度 2 mg/ml，其中在藥物釋放的過程中搭配劑量 10 Gy 的放射線治療，動物組別分為為治療組、水膠組、卡鉑組、卡鉑水膠組、放射組、卡鉑放射組、卡鉑水膠放射組，其中以卡鉑放射組、卡鉑水膠放射組具有治療的效果並將存活率提高至百分之五十。
綜合所述的實驗結果，比較了各組不同治療的方法，最後比較了卡鉑放射組與卡鉑水膠放射這兩種方法，卡鉑水膠簡化對小鼠神經膠質瘤的治療且同時保有放射協同效應，提供了術後注射後照放射的臨床應用。

High grade glioma is the most common aggressive type of brain tumors. According to statistics, 3 ~ 5 cases occur per 100,000 population per year. Even if the patients receive a variety of treatments, their survival rate is relatively low. The main reason is that glioma has diffuse infiltration properties that grow outwardly into normal brain tissue leading to a higher recurrence of the prognosis; hence, the objective of this study is to offer a way that may effectively inhibite tumor growing and prolong survival.
In the study, oxidized hyaluronic acid hydrogel as carboplatin carrier with radiotherapy are used to ttreat for mice with glioma, to evaluate the efficacy of the other animal experiments, and to improve Gliadel® Wafer problems by providing new options for treatment.
The results show that the modified oxidized hyaluronic acid and adipic acid dihydrazide form hydrogel, and its functional group change is evident through the application of FTIR. In addition, it was found carboplatin exsisted in the hydrogel by the EDS element analysis. In the rheometer analysis, we could utilize temperature to control gelling time. Moreover, the release profile and treatment schedule that the drug release with radiation therapy are consistent. In the treatment on mice with glioma, the survival rate increased to 50 percent.
In conclusion, comparing with carboplatin and hydrogel carboplatin simplified the drug delivery for mice gliomas and remained the synergic effect with radiotherapy, which provides clinical application of intraoperative injection followed by radiotherapy.

致謝 i
中文摘要 ii
圖目錄 vii
表目錄 ix
第一章 前言 1
1.1 神經膠質瘤 1
1.2 神經膠質瘤的現今治療 3
1.2.1 手術治療 3
1.2.3 化學藥物治療 4
1.3 血腦屏障 ( blood-brain-barrier, BBB ) 5
1.4 局部藥物釋放 6
1.5 研究目的 9
第二章 理論基礎 10
2.1 卡鉑 10
2.2 高分子水膠做藥物載體 11
2.2.1 溶劑交換 11
2.2.2 光聚合反應 11
2.2.3 離子鍵交聯 11
2.2.4 pH值成膠 11
2.2.5 溫度成膠 12
2.3 透明質酸（Hyaluronic acid, HA） 13
2.4 氧化透明質酸 ( Oxidized hyaluronic acid, Oxi-HA )……....………….…..14
2.5 水膠的流變特性 16
第三章 材料與方法 17
3.1 實驗儀器與藥品 17
3.2 實驗流程與方法 20
3.3 實驗流程圖 20
3.4 材料製備 21
3.4.1 氧化透明質酸製備 21
3.4.2 氧化透明質酸/己二酸二醯肼/卡鉑水膠製備 21
3.5 材料分析 22
3.5.1 傅立葉轉換紅外線分析 22
3.5.2 氧化程度 23
3.5.3 能量散射光譜儀 24
3.5.4 流變儀分析 25
3.5.5 材料降解 25
3.5.6 藥物釋放曲線 25
3.6 生物安全性測試 26
3.6.1 材料的細胞毒性分析 26
3.6.2 卡鉑之安全劑量 31
3.7 動物實驗 32
3.7.1 體內動物模式 32
3.7.2 生物冷光影像分析 34
第四章 結果與討論 35
4.1 材料分析 35
4.1.1 傅立葉轉換紅外線分析 35
4.1.2 TNBS assay 偵測氧化程度 37
4.1.3 EDS元素分析 38
4.1.4 流變性質 39
4.1.5 材料降解 40
4.1.6 藥物釋放 41
4.2 生物相容性 42
4.2.1 WST-1 assy 42
4.2.2 LDH assay 43
4.2.3 Live & Dead staining 44
4.2.4 半抑制濃度 ( half maximal inhibitory concentration, IC50 ) 46
4.3 動物實驗 47
4.3.1 體重分析 47
4.3.2 腫瘤體積分析 48
4.3.3 冷光分析 49
4.3.4 存活分析 52
4.3.5 組織切片分析 53
4.3.6 血液分析 56
第五章 結論 57
第六章 參考文獻 58

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