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    請使用永久網址來引用或連結此文件: http://libir.tmu.edu.tw/handle/987654321/50832


    題名: 含茶多酚奈米載體於乾眼症治療之研究
    Preparation and Evaluation of Tea Polyphenol contained Nanocarriers for Dry Eye Syndrome Treatment
    作者: 邱紋瑛
    Chiu, Wen-Ying
    貢獻者: 生醫材料暨組織工程研究所
    曾靖孋
    楊維中
    關鍵詞: 表沒食子兒茶酸鹽;明膠;玻尿酸;奈米載體;乾眼症
    (-)-Epigallocatechin-3-gallate;Gelatin;Hyaluronic acid;nanoparticles;Dry Eye Syndrome
    日期: 2013-07-16
    上傳時間: 2018-10-03 17:25:32 (UTC+8)
    摘要: 乾眼症在眼科臨床上，定義為眼睛表面發炎的一種疾病，發炎因子會刺激上皮細胞、杯狀細胞及神經等，使淚膜產生不穩定以及淚液高滲透壓等情形。在過去乾眼症好發於老年人口，但隨著科技的進步，過度使用電子產品加上用眼習慣不良的因素，患者年齡層快速下降且罹患人口增高。眼藥水是眼科用藥常見的劑型，但給藥後卻只有低於5%的藥量會停留在眼部組織，生物利用率非常的低。為了上發展一個更有效的眼科用藥，本研究欲製備黏附型高分子奈米載體，藉由黏附型高分子改質，提高載體與淚液層、角膜的交互作用，進而增加奈米載體在眼部表面所停留的時間，提高其生物利用率。
    表沒食子兒茶酸鹽(EGCG)為茶多酚中主要成分之一，其具有抗氧化及抗發炎反應之特性，故針對乾眼症的發炎症狀，本研究選用此藥物進行治療評估。明膠(Gelatin)為生物性高分子，作為藥物載體有良好的生物相容性。而玻尿酸(HA)可增加載體及角膜接觸時的黏附性，故結合上列三種材質進進行奈米藥物載體研製。以期延長載體於角膜停留的時間，增加藥物的生物利用率，達到治療乾眼症效果。
    藉由各種製備參數的探討，本研究所得製備之奈米載體最佳化條件為： GNPs-GE125H (Gelatin-EGCG-HA混合比例為2 mg/ml-2 mg/ml-125 μg/ml)；其粒徑大小為222.90 ± 1.73 nm，表面電荷為12.19 ± 0.34 mV，且EGCG之包覆率可達99.56 %。在穿透式電子顯微鏡下觀察，顯示載體呈圓球狀且均勻分布，亦可清楚觀察到HA在載體外圍的結構。以螢光染劑模擬EGCG釋放，奈米載體GNPs-GE125H之螢光相較於螢光水溶液更為緩慢釋放。進而以人類角膜上皮細胞共培養，所有組別在EGCG濃度20 μg/ml以下，均有良好的細胞活性，但至第七天時EGCG水溶液對細胞活性沒有影響，GNPs-GE以及GNPs-GE125H對細胞依然有持續性影響力，顯示兩者均有緩釋效果；且GNPs-GE125H更為明顯。在螢光奈米載體與細胞共培養的的實驗中，顯示GNPs-GE125H螢光奈米載體於細胞內之螢光含量較高，亦證實GNPs-GE125H螢光奈米載體可達到細胞內較高濃度累積。此外將細胞進行氧化破壞再給予奈米載體共培養實驗中，顯示EGCG具有降低氧化傷害的效果。根據上列實驗成果，顯示本研究所研製之Gelatin-EGCG-HA奈米載體具有良好生物相容性、藥物緩釋效果；並可提高載體於角膜細胞累積量、且可降低角膜細胞氧化傷害程度，因此具有應用於乾眼症治療之可行性。
    Aging population has become more and more percentage. With long time watching computer or phone, these also cause significant increase of patients who get dry eye Syndrome (DES). DES is a general disease in ophthalmic clinic causing by ocular surface inflammation. The major treatment for DES is using artificial tears and eye-drop contained hyaluronic acid (HA). But those just only help maintain moisture on the ocular surface, no anti-inflammation effect. In ophthalmological treatment, therapeutic agent is usually quickly removed to nasolacrimal duct and is actually hampered by the lack of an efficient ocular drug delivery carrier.
    Nanocarries offers a promising alternative chance for the treatment of ocular diseases to do the possibility to increase drug reach in the ocular surface. The main objective of the present work is to fabricate bioadhesive polysaccharides, hyaluronic acid (HA) and biodegradable polymer, gelatin, intended for the delivery of drug to the anterior ocular tissue especially cornea. In this study, the flavonoid, (-)-epigallocatechin gallate (EGCG), was chosen for its anti-inflamation ability for treat.
    The particle size of GNPs-GE125H (2 mg/ml Gelatin-2 mg/ml EGCG-125 μg/ml HA) was found to be of nanometric size at 222.90 ± 1.73 nm with positive zeta potential at around 12.19 ± 0.34 mV. EGCG encapsulation rate were about 99.56 %, and has slow release effect. A round shape of the nanoparticles was observed from TEM image. From the release rate experiment, solution from of fluorescence was quickly release in to PBS, but GNPs-GE and GNPs-GE125H showed slow release behavior. EGCG, GNPs-GE and GNPs-GE125H solution at concentration 20 μg/ml of EGCG were non-toxic with HCE-T cell. The GNPs-GE and GNPs-GE125H solution could caused lower effect cell viability, on day 7. From anti-oxidation experiment, cell treated with EGCG, GNPs-GE and GNPs-GE125H solution showed higher cell viability than H2O2 treated one. This indicated that these EGCG formulations maintain the anti-oxidation of EGCG.
    In conclusion, EGCG could be encapsulated in biological polymer-gelatin and HA to from as cationic nanoparticles; it may have potential to inter adsorbed on negatively charged cornea for ocular drug delivery. So we expect this bioadhesive nanocarriers (GNPs-GE125H) have good treatment effect of DES patients.
    描述: 碩士
    指導教授-曾靖孋
    共同指導教授-楊維中
    委員-吳其昌
    委員-董國忠
    委員-謝明發
    資料類型: thesis
    顯示於類別:[生醫材料暨組織工程研究所] 博碩士論文

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