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標題: | 幾丁聚醣奈米粒子包載靛藍二磺酸鈉以偵測大腸癌細胞 Chitosan Nanoparticles Loaded with Indigo Carmine for Endoscopy Detection of Colorectal Cancer cells |
作者: | Jian-Wen Chen 陳建文 |
指導教授: | 謝銘鈞(Ming-Jium Shieh) |
關鍵字: | 幾丁聚醣,靛藍二磺酸鈉,葉酸, chitosan,indigo carmine,folic acid, |
出版年 : | 2007 |
學位: | 碩士 |
摘要: | 在大腸直腸癌的診斷中,內視鏡是一項方便且普遍使用的診斷工具,由內視鏡所觀察到的影像可以判斷腸子的健康情況。而為了彌補純目視下的不足,有兩種方式可以作為輔助的判斷。一種為染色內視鏡,另一種為放大內視鏡。前者為使用染劑(如indigo carmine)直接噴灑在腸壁的表面,增加層次性,並且會容易累積在潰瘍、糜爛部位的底部,摺疊處、凹陷處等,可將病灶與周邊正常黏膜的界限區分。而後者為可以將倍率放大至一百倍,已經是準微觀(semi-micro)的程度,可以直接對病灶處看的更為仔細。
但indigo carmine並無特定標靶的能力,因此本研究目的為合成一表面接枝上具有特定標靶到癌細胞能力的配體(如葉酸)的幾丁聚醣高分子,並且把接枝上葉酸的幾丁聚醣高分子利用離子凝膠法與藍色染劑Indigo carmine製備成奈米粒子,使其奈米粒子同時具有特定標靶到癌細胞以及顯色的功用。幾丁聚醣為生物可分解、生物可相容性、無毒、抗菌的高分子材料。把接枝上葉酸的幾丁聚醣與indigo carmine製備成的奈米粒子,量測其粒徑大小、表面膜電位、和包載indigo carmine的效率。幾丁聚醣奈米粒子標靶到細胞的能力的評估則是把含有奈米粒子的培養基直接加在HT-29細胞上,經過一段時間後,來量測其粒子黏覆在細胞的量。 由實驗結果得知,所製備的幾丁聚醣奈米粒子 (包載0.5 mg/mL indigo carmine;0.02N acetic acid) 的平均粒徑介於120~140nm之間,並隨著葉酸改質率提高而使得尺寸也稍稍變大。表面膜電位則大約略大於20mV。而細胞黏附幾丁聚醣奈米粒子的累積量隨著時間增加,其中葉酸改質率較高的幾丁聚醣所製備的奈米粒子具有較好的效果。顯示奈米粒子接枝上較多的葉酸對HT-29細胞具有較好的標靶效果。 綜合上述,本實驗針對包載indigo carmine 的幾丁聚醣奈米粒子的特性進行體外研究的探討,由這些結果將有助於包載indigo carmine的幾丁聚醣奈米粒子於大腸直腸癌上用內視鏡診斷上的應用。 Endoscopy is a common diagnosis device for the detection of colorectal cancer. From the view of bowel wall we could determine the condition of intestine. There are two methods to improve the diagnostic sensitivity. One is chromoendoscopy, and the other is magnifying endoscopy. The former uses color contrast dye such as indigo carmine to spray on the surface of intestine which can accumulate in the pits and crevices of the mucosa to identify the lesion. The latter could magnify until one hundred times, and watch clearly. But indigo carmine does not target specifically. Due to the reason we want to synthesis chitosan conjugated with target-specific ligand such as folic acid and use this polymer to prepare chitosan nanoparticles loaded with indigo carmine by ionic gelation method. Let these particles combine the targeting ability and contrast stain effect. Chitosan is biodegradable, biocompatible, nontoxicity, and antibacterial. In this study, the particle size, zeta potential and loading efficiency were determined. Moreover, the targeting effect of chitosan nanoparticles was detected by adding them into the medium directly to incubate the colon cell line HT-29 cells. The particle size of chitosan nanoparticles ( loaded with 0.5 mg/mL indigo carmine;0.02N acetic acid ) were between 120nm and 140nm, and increased gradually with the rise of conjugated ratio of folic acid. The zeta potential is about 20mV. The accumulation of cell adhesive increased with incubation time, and the higher conjugated ratio of folic acid reveal better effect. The result shows chitosan conjugated with more folic acid could possess good targeting efficiency in HT-29 cells. To sum up, the characterizations of chitosan nanoparticles loaded with indigo carmine in vitro were studied. These results may improve the diagnosis of colorectal cancer through endoscopy by chitosan nanoparticles loaded with indigo carmine. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/28004 |
全文授權: | 有償授權 |
顯示於系所單位: | 醫學工程學研究所 |
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