於本篇研究中，透過化學共沉澱法成功的合成出具有介孔結構的磁性氫氧基磷灰石奈米顆粒(Mesoporous magnetic hydroxyapatite, MPmHAp)。因MPmHAp具超順磁性且含獨特的介孔結構，如高的表面積(54 m2/g)，於藥物釋放實驗中證明此搭載抗癌藥物Doxorubicin的HER2-MPmHAp能有效的控制並延長藥物釋放速率(130 hr)，說明此HER2-MPmHAp能搭載更大量的藥物分子及具有緩慢釋放的特性，適合作為藥物載體之模型。進一步將合成的MPmHAp表面接枝HER-2抗體(HER2-MPmHAp)來達到標靶乳癌細胞的目的。過度表現HER-2受體的腫瘤細胞透過受體介導的內吞作用(receptor mediated endocytosis)具大量的奈米顆粒含量(341.1 pg/cell)。最後，透過搭載抗癌藥物及藉由外加磁場的作用下，針對腫瘤細胞達到熱化療的雙重療效。

In this study, the result demonstrated that iron doped hydroxyapatite nanocrystal with mesoporous structure was successfully prepared through one step route. mesoporous materials have gained enhanced interest with particular attention as drug storage and release hosts due to their higher surface area(54 m2/g) and unique textural properties. This material exhibits rod-like, crystalline structure, which is suitable for drug (Doxorubicin) release as drug carrier and prolong the release time(130 hr).Due to some tumor may overexpressing HER-2 genes, so it can immobilization HER-2 antibody onto MPmHAp surface to active targeting tumor cell and through receptor-mediated endocytosis, it can enhance cell uptake of HER2-MPmHAp(341.1 pg/cell). Finally, it can using chemohyperthermia that hyperthermia improves the antitumor effect of some chemotherapeutic agents.

圖目錄 VII
表目錄 IX
第一章 緒論 1
1.1 前言 1
1.2 乳癌 3
1.3 腫瘤之臨床治療 5
1.3.1 手術 5
1.3.2 化學療法 6
1.3.3 放射療法 7
1.3.4 熱治療 8
1.4 乳癌治療方法 10
1.5 研究目的 11
第二章 文獻回顧 13
2.1 超順磁奈米顆粒於癌症治療的應用 13
2.1.1 磁性產熱的機制 14
2.1.2 磁性氫氧機磷灰石(mHAp) 16
2.2 磁性奈米晶體的表面修飾 18
2.2.1 聚乙二醇(PEG) 19
2.3 介孔(mesoporous) 21
第三章 實驗與方法 23
3.1 實驗流程圖 23
3.2 實驗步驟與方法 25
3.2.1製備介孔磁性氫氧基磷灰石( MPmHAp) 25
3.2.2介孔磁性氫氧基磷灰石之表面改質( APTES-MPmHAp) 26
3.2.3接枝HER-2抗體( HER2-MPmHAp) 27
3.3 材料分析 28
3.3.1 X光繞射分析(X-Ray Diffractometer, XRD) 28
3.3.2 感應耦合電漿光學放射光譜(Inductively Coupled Plasma, ICP) 29
3.3.3 穿透式電子顯微鏡(Transmission Electron Microscopy, TEM) 30
3.3.4 震動樣品磁力計(Vibrating Sample Magnetometer, VSM) 31
3.3.5 高周波電感應加熱器(High Frequency-Induction Heating machine, HFIHM) 32
3.3.6 BET比表面積測試(Brunauer-Emmett-Teller, BET) 33
3.3.7 體外(in vitro)藥物裝載及釋放 34
3.4 細胞培養及生物相容性測試 35
3.4.1 細胞培養及計數 36
3.4.2 LDH細胞毒性測試 38
3.4.3 細胞攝取定量分析 40
第四章 結果與討論 41
4.1 介孔磁性氫氧基磷灰石之物化性分析 41
4.1.1 X光繞射分析 41
4.1.2穿透式電子顯微鏡 43
4.1.3 比表面積測試 46
4.1.4 元素分析 48
4.1.5 震動樣品磁力計 50
4.1.6高周波電感應加熱器 52
4.1.7藥物裝載及釋放 53
4.2 體外(in vitro)細胞分析 55
4.2.1細胞毒性 55
4.2.2細胞攝取之定量分析 57
4.2.3 細胞攝取之毒性測試 60
第五章 結論 62

圖目錄
圖 1- 1 抗體的基本構造 4
圖 1- 2 實驗設計圖 12

圖 2- 1 Neel自旋及布朗運動 15
圖 2- 2 超順磁材料與外加磁場關係 15
圖 2- 3 氫氧基磷灰石的分子結構圖 17
圖 2- 4聚乙二醇之化學結構式 20
圖 2- 5介孔(mesoporous)示意圖 22

圖 3- 1 LDH反應過程 38

圖 4- 1 不同鐵濃度含量的MPmHAp之XRD分析 42
圖 4- 2 不同PEG濃度的MPmHAp之XRD分析 42
圖 4- 3 MPmHAp 21之TEM影像，箭頭處為氧化鐵 44
圖 4- 4 MPmHAp之HR-TEM影像 44
圖 4- 5 MPmHAp之電子擇區繞射圖 45
圖 4- 6 (a) MPmHAp 21及(b) MPmHAp 24之BET曲線圖及BJH之分佈圖 47
圖 4-7 合成的MPmHAp在不同鐵濃度下之鈣(●)及鐵(□)含量 49
圖 4- 8 (a) MPmHAp 23及(b) MPmHAp 24於30kOe下之VSM分析 50
圖 4- 9 (a) MPmHAp 24及(b) MPmHAp 34於30kOe下之VSM分析 51
圖 4- 10外加交變磁場作用下，0.5 mg/ml的(a) MPmHAp24及(b) MPmHAp 34於水中之溫度變化量 52
圖 4-11 DOX-HER2-MPmHAp 24於pH7.4之藥物釋放曲線圖 54
圖 4- 12 3T3細胞與不同濃度的MPmHAp 4共培養24hr及72hr之細胞毒性分析 56
圖 4- 13 低表現HER-2受體的乳腺癌細胞MCF-7及過度表現HER-2受體的乳腺癌細胞SK-BR-3與HER2-MPmHAp共培養至12小時之定量分析 59
圖 4- 14 搭載抗癌藥物Doxorubicin之HER2-MPmHAp奈米顆粒藉由外加磁場作用下之細胞毒性測試 61

表目錄
表 1 合成MPmHAp所含不同莫耳量的聚乙二醇(PEG)及氧化鐵(Iron oxide)對照表 24
表 2 MPmHAp質地結構之特性分析（SBET:Surface area, Vp:Pore volume, Dp:Pore size） 47
表 3添加不同鐵源前驅物MPmHAp之成分元素分析（*理論值） 48

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