| 摘要: | 本研究利用幾丁聚醣(chitosan, CS )奈米粒子作為多酚化合物(polyphenol)-没食單寧(gallotannin, GT)之載體(carrier)，利用三聚磷酸鈉(tripolyphosphate , TPP)與幾丁聚醣之間電荷反應，以離子凝膠技術(ionic gelation)包埋(entrapped)或吸附(adsorbed)没食單寧以及使用多層乳化包覆技術(water-oil-water)包覆(encapsulated)没食單寧的方式製備出一系列没食單寧-幾丁聚醣奈米粒子。比較不同製備方法的奈米粒子：包埋、吸附與包覆之間的差異性。利用穿透式電子顯微鏡(Transmission electron microscopy , TEM)和動態光散射粒徑分析儀(Dynamic light scattering, DLS)分別測試奈米粒子的直徑大小、界面電位(Zeta potential)及形狀外貌…等；並針對影響奈米粒子的物化性質作探討。其奈米粒子的粒徑大小和承載沒食單寧的包覆率，皆會因不相同的製備過程而有不同地影響因素。測試没食單寧-幾丁聚醣奈米粒子對纖維母細胞(L929 mouse fibroblast)的活性測試以及對革蘭氏陽性菌(Gram-positive coccus)—金黃色葡萄球菌(Staphylococcus aureus)之抗菌活性，藉以量測出最小抑菌濃度(minimum inhibitory concentration, MIC)及最小殺菌濃度(minimum bactericidal concentration, MBC)。研究結果發現將沒食單寧包裹於幾丁聚醣奈米粒子中，顆粒大小圓潤、平均，其中更以W/O/W製備得之結果為最佳；在一般環境下，奈米粒子能夠保護核蕊物質--單寧，不會因受到外在環境而產生變化；而是在給予一個特定仿造生物體環境之條件下，例如：pH値、機械性質…等，造成奈米粒子的幾丁聚醣外殼會溶解破裂，使沒食單寧逐漸釋放出來。沒食單寧無細胞毒性但奈米粒子中的單寧濃度會對細胞生長造成影響，而其粒子對金黃色葡萄球菌具有抗菌活性。
This study presents the preparation and characteristic of chitosan (CS) nanoparticles as a carrier of polyphenol compound – gallotannin (GT). The preparation method of CS nanoparticles is using the electric charge interaction between sodium tripolyphosphate (TPP) and chitosan. Gallotannin is entrapped or absorbed by the ionic gelation technology with chitosan as well as using multi-layer emulsification encapsulation technology: water-oil-water encapsulation gallotannin to prepare a series of GT-CS nanoparticles. We also discuss the differences of these three nanoparticle preparation methods (entrapped, absorbed, encapsulated ).
The nanoparticles’ diameter, zeta potential and the shape are measured by using transmission electron microscopy (TEM) and the dynamic light scattering (DLS). The influences on the physicochemical characterization of nanoparticles are also discussed. The cell’s activity with L929 mouse fibroblast and antibacterial activity with the Gram-positive coccus-Staphylococcus aureus of GT-CS nanoparticle were detected, and minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) are measured. Those results show that gallotannin had encapsulated in chitosan nanoparticle, and the particle was round and size average, especially the W/O/W particles had the best effect. The nanoparticles could protect the core material-tannin under the normal environment. But in the specific condition such as creating the outer covering, chitosan nanoparticle could be dissolved and cause gallotannin to release gradually under the copy organisam environment, for example: pH, mechanical property, etc. Gallotannin is found non-toxicity but the tannin density in nanoparticle would influence on the cell growth. These particles had the antibacterial activity to Staphylococcus. |
