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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 李伯訓 | |
dc.contributor.author | Ren-Yin Chang | en |
dc.contributor.author | 張人尹 | zh_TW |
dc.date.accessioned | 2021-06-17T02:16:48Z | - |
dc.date.available | 2021-02-22 | |
dc.date.copyright | 2018-02-22 | |
dc.date.issued | 2017 | |
dc.date.submitted | 2017-09-22 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/68292 | - |
dc.description.abstract | 順鉑(cisplatin)用來治療口腔癌已超過三十幾年,不過嚴重的副作用限制順鉑在癌症的治療上的應用,然而順鉑透過奈米粒子的攜帶可以降低在使用上所帶來的毒性。因此本研究利用兩種不同材料-幾丁聚醣(chitosan)聚乳酸聚甘醇酸(poly(lactic-co-glycolic acid, PLGA)以及普魯士藍製備奈米粒子,觀察攜帶順鉑之效果,同時利用離子導入法(iontophoresis)加強奈米粒子穿透癌症部位的能力。合成之奈米粒子透過FTIR (Fourier transform infrared spectroscopy)分析官能基,動態光散射分析儀(Dynamic Light Scattering)檢測粒徑大小,分布程度,穿透式電子顯微鏡(Transmission electron microscope)觀察顆粒大小、分散性、一致性、粒子型態。在藥物包覆方面,使用可見光紫外光分光光譜儀(UV-vis)製作檢量線並測量包覆率、釋放率。體外實驗則使用兩株不同口腔癌細胞SAS、Cal27進行模擬,經過細胞活性檢測(MTT assay) 觀察奈米粒子本身之生物相容性以及藥物釋放後癌細胞毒殺之效果。
實驗結果顯示,本研究第一部分合成之幾丁聚醣-聚乳酸聚甘醇酸奈米粒子在FTIR檢測出聚乳酸聚甘醇酸本身之特徵峰1082 cm−1(C-O-C)以及1749 cm−1 (C=O stretching),同時也觀察到奈米粒子外包覆的幾丁聚醣特徵峰3332 cm−1(NH2, OH)、1550 cm−1 (amide I N-H bending vibration),顯示幾丁聚醣成功包覆在奈米粒子外。穿透式顯微鏡下則觀察到100~150nm範圍的粒徑大小以及圓形的顆粒形狀。在藥物包覆率方面原聚乳酸聚甘醇酸奈米粒子為3.08±0.02%,而幾丁聚醣-聚乳酸聚甘醇酸奈米粒子則無法成功包覆藥物。實驗第二部分合成之普魯士藍奈米粒子,FTIR檢測當中確實有出現普魯士藍的特徵峰2064cm−1 (C-N stretching),以及包覆層PVP的特徵峰1641cm−1碳氧雙鍵(amide II C=O stretching)。在穿透式電子顯微鏡下粒子大小分布為100nm,型態為方形。普魯士藍奈米粒子順鉑包覆率為39.85±7.83%,藥物釋放方面可持續釋放二天,釋放量達43.20± 5.55μg。在細胞毒殺試驗中,普魯士藍奈米粒子250μg/ml組可使SAS及Cal27兩株癌細胞的存活率分別下降至5.95±0.37%以及14.57±1.26%,展現出良好的抗癌效果。在iontophoresis導入實驗中,可以觀察到iontophoresis成功驅動普魯士藍奈米粒子穿透纖維通透膜。 | zh_TW |
dc.description.abstract | Cisplatin has been used for treating oral cancer for over 3 decades, but severe side effects limit its application. Nanoparticles were synthesized to decrease toxicity of cisplatin. The aim of this study was to prepare nanoparticles by chitosan-PLGA (poly (lactic-co-glycolic acid) and prussian blue and observe the encapsulation efficiency of two different materials. The iontophoresis was used to enhance the penetrating ability of nanoparticles on tumor site. Functional group of nanoparticles were measured by FTIR (Fourier transform infrared spectroscopy). The nanoparticles size and morphology were analyzed by DLS (Dynamic Light Scattering) and TEM (Transmission electron microscope). UV-VIS spectrophotometer was used to determine the encapsulation efficiency and release rate of cisplatin. The anticancer efficacy of nanoparticles on SAS and Cal27 were evaluated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay.
In the first part, FTIR results of chitosan-PLGA nanoparticles showed the characteristic peaks of 1082 and 1749 cm−1 which were identified as stretching vibrations of the C-O-C and C=O groups in PLGA, respectively. The 3332 cm−1 was the characteristic peak of N-H and O-H stretching vibrations in chitosan. The morphology of nanoparticles were spherical and the size of particles ranged between 100-150nm under TEM. PLGA nanoparticles showed the encapsulation efficiency as 3.08±0.02%, but the cisplatin could not be successfully entrapped to chitosan-PLGA-cisplatin nanoparticles. For the second part of result, FTIR demonstrated that prussian blue had characteristic peak at 2064 cm-1 associated with C-N functional group. The characteristic peak of PVP was also observed at 1641cm-1 as stretching vibrations of C=O group. The TEM indicated that Prussian blue particles were cubic shape as well as the size of particles were around 100nm. Encapsulation efficiency of prussian blue nanoparticles was calculated as 39.85±7.83%. The total amount of drug released from prussian blue nanoparticles was 43.20± 5.55μg. The cell viability of SAS and Cal27 were decreased to 5.95±0.37% and 14.57±1.26% respectively after incubation with 250μg/ml prussian blue nanoparticles for 3 days. After applying iontophoresis for 3 hours, the prussian blue nanoparticles successfully penetrated the cellulose membrane. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T02:16:48Z (GMT). No. of bitstreams: 1 ntu-106-R04450009-1.pdf: 5094563 bytes, checksum: f9253a48b62edcd8db06b48b21cae55b (MD5) Previous issue date: 2017 | en |
dc.description.tableofcontents | 口試委員會審定書 i
致謝 ii 摘要 iii ABSTRACT v 目錄 vi 圖目錄 ix 表目錄 xi Chapter 1 緒論 1 Chapter 2 文獻回顧 2 2.1 口腔癌 2 2.2 順鉑(cisplatin) 2 2.3 奈米粒子 3 2.4 高分子聚合奈米粒子 3 2.5 幾丁聚醣 4 2.5.1 幾丁聚醣黏膜附著性(Mucoadhesive) 5 2.5.2 Tight junctions 5 2.6 PLGA 6 2.6.1 雙重乳化法(double emulsion) 7 2.7 配位聚合物(Coordination polymers) 8 2.7.1 普魯士藍 9 2.7.2 孔洞型普魯士藍(Porous Prussian blue) 9 2.8 離子導入法 10 Chapter 3 材料方法 11 3.1 實驗藥品 11 3.2 實驗儀器 13 3.3 實驗流程圖(圖 3 1) 15 3.4 奈米粒子製備 16 3.4.1 幾丁聚醣聚乳酸聚甘醇酸(Chi-PLGA)奈米粒子製備 16 3.4.2 普魯士藍(Prussian Blue)奈米粒子製備 19 3.5 奈米粒子基本特性檢測 20 3.5.1 動態光散射分析儀(Dynamic Light Scattering, DLS) 20 3.5.2 傅立葉轉換紅外線光譜儀(FTIR) 20 3.5.3 穿透式電子顯微鏡(Transmission electron microscope, TEM) 20 3.5.4 掃描式電子顯微鏡(Scanning Electron Microscope) 21 3.6 奈米粒子藥物包覆率及控制釋放 21 3.6.1 Cisplatin檢量線製備 21 3.6.2 PLGA奈米粒子藥物包覆率、藥物承載率計算 22 3.6.3 普魯士藍奈米粒子藥物包覆率、藥物承載率計算 22 3.6.4 普魯士藍奈米粒子藥物釋放 23 3.7 細胞培養及藥物測試 23 3.7.1 細胞培養液配置 23 3.7.2 解凍細胞 24 3.7.3 繼代培養 24 3.7.4 冷凍細胞 24 3.8 細胞活性測試(MTT Assay) 25 3.8.1 普魯士藍奈米粒子生物相容性測試 25 3.8.2 普魯士藍奈米粒子抗癌效果測試 25 3.9 Iontophoresis導入奈米粒子 26 Chapter 4 實驗結果 27 4.1 Chi-PLGA奈米粒子特性分析 27 4.1.1 FTIR 27 4.1.2 動態光散射分析儀(DLS) 27 4.1.3 穿透式電子顯微鏡(TEM) 28 4.2 PLGA藥物包覆率變因探討 28 4.2.1 不同PVA對藥物包覆影響之探討 28 4.2.2 不同藥物量對藥物包覆影響之探討 28 4.2.3 不同高分子濃度對藥物包覆影響之探討 29 4.2.4 幾丁聚醣對藥物包覆影響之探討 29 4.3 普魯士藍奈米粒子特性分析 29 4.3.1 FTIR 29 4.3.2 動態光散射分析儀(DLS) 30 4.3.3 穿透式電子顯微鏡(TEM) 30 4.3.4 掃描式電子顯微鏡(SEM)及能量散射X-射線光譜(EDS) 30 4.4 普魯士藍奈米粒子藥物包覆率及釋放 31 4.4.1 PB-Cis藥物包覆率及藥物承載率 31 4.4.2 PB-Cis藥物控制釋放 31 4.5 普魯士藍奈米粒子於細胞釋放之情形 31 4.5.1 PB-shell生物相容性測試 31 4.5.2 PB-Cis細胞毒殺效果 32 4.6 Iontophoresis傳遞普魯士藍奈米粒子之效果 32 Chapter 5 討論 57 5.1 PLGA奈米粒子大小及型態 57 5.2 PLGA奈米粒子包覆率變因探討 57 5.2.1 PVA對包覆率之影響 58 5.2.2 藥物量對包覆率之影響 58 5.2.3 高分子聚合物濃度對包覆率之影響 58 5.2.4 幾丁聚醣對包覆率之影響 59 5.3 普魯士藍奈米粒子大小及型態 59 5.4 普魯士藍奈米粒子藥物包覆及控制釋放 60 5.5 普魯士藍奈米粒子細胞測試 61 5.5.1 生物相容性 61 5.5.2 PB-Cis癌細胞毒殺試驗 61 5.6 Iontophoresis傳遞普魯士藍奈米粒子 62 Chapter 6 結論 63 | |
dc.language.iso | zh-TW | |
dc.title | 攜帶順鉑之奈米粒子對口腔癌的治療及應用 | zh_TW |
dc.title | Development of Cisplatin Loaded Nanoparticles for
Treating Oral Cancer | en |
dc.type | Thesis | |
dc.date.schoolyear | 106-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 張哲政,魏大欽 | |
dc.subject.keyword | 奈米粒子,聚乳酸聚甘醇酸,幾丁聚醣,普魯士藍,順鉑,口腔癌,離子導入法, | zh_TW |
dc.subject.keyword | oral cancer,PLGA,chitosan,prussian blue,cisplatin,nanoparticles,iontophoresis, | en |
dc.relation.page | 70 | |
dc.identifier.doi | 10.6342/NTU201702566 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2017-09-25 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 口腔生物科學研究所 | zh_TW |
顯示於系所單位: | 口腔生物科學研究所 |
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