請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/68292
標題: | 攜帶順鉑之奈米粒子對口腔癌的治療及應用 Development of Cisplatin Loaded Nanoparticles for Treating Oral Cancer |
作者: | Ren-Yin Chang 張人尹 |
指導教授: | 李伯訓 |
關鍵字: | 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, oral cancer,PLGA,chitosan,prussian blue,cisplatin,nanoparticles,iontophoresis, |
出版年 : | 2017 |
學位: | 碩士 |
摘要: | 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。 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. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/68292 |
DOI: | 10.6342/NTU201702566 |
全文授權: | 有償授權 |
顯示於系所單位: | 口腔生物科學研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-106-1.pdf 目前未授權公開取用 | 4.98 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。