本研究利用雙碳鏈陽離子型界面活性劑dihexadecyldimethyl-ammonium bromide （DHDAB）及單碳鏈陰離子型界面活性劑sodium dodecylsulfate （SDS）所形成之類三碳鏈離子對雙親分子dihexadecyldimethylammonium-dodecylsulfate （DHDA-DS），混合膽固醇以強制型製程製備出陰陽離子液胞，並分析膽固醇對於液胞物理特性、雙層膜的相轉移行為及流動性的影響。此外，也探討液胞分散液稀釋後的物理穩定性，並進一步分析其細胞活性。實驗結果顯示，純成分DHDA-DS可形成液胞，但可能由於水溶性較佳的DS-易脫離雙層膜結構，導致液胞帶正電，由於有足夠的帶電性使液胞具有良好的物理穩定性。當膽固醇添加量高於20 mol%後，界面電位開始些微上升，推測是雙層膜結構中開始有膽固醇結晶產生，其固醇環對DS-從雙層膜溶出的抑制能力降低，進而影響雙層膜的排列行為。透過螢光偏極化法及動態雷射光散射法分析雙層膜的相轉移行為，添加10 mol%膽固醇時讓雙層膜排列更緊密，導致相轉移溫度增加幅度較顯著。當膽固醇添加量高於20 mol%後，液胞雙層膜的相轉移行為幾乎消失。由紅外光譜法及螢光偏極化法分析雙層膜的流動性，在膽固醇結晶生成前，雙層膜整體的流動性無明顯的變化，但靠近雙層膜中心的流動性上升，造成這樣的差異可能是膽固醇結構對不同碳氫鏈區段誘導出不同效應所導致。當膽固醇結晶生成後，整體碳鏈的流動性上升，由膽固醇的不規則效應主導，但靠近雙層膜中心的流動性則因為螢光分子轉動的空間減少而下降。稀釋後液胞的界面電位隨著稀釋倍率的增加呈現先升後降的趨勢，但帶電量依舊足夠提供液胞良好的物理穩定性。利用穩定性足夠的液胞進行正常人類支氣管上皮細胞的活性檢測，由於在純水環境下製備出的陰陽離子液胞在培養液中容易有聚集沉澱，導致無法得知準確的細胞毒性，因此將製備陰陽離子液胞的環境置換成體積比14／1的水與DMSO混合液，目的是為了增加液胞分散液與培養液間的作用，減緩沉澱現象。由顯微鏡觀察，當液胞分散液稀釋至0.25 mM時，和純水環境下製備並稀釋至0.2 mM相比，黑色聚集沉澱有顯著的減少，藉由存活率的計算也發現膽固醇的添加及稀釋倍率的提高均有助於減緩液胞毒性。

In this study, pseudotriple-chained ion pair amphiphile, dihexadecyldimethylammonium-dodecylsulfate (DHDA-DS), was prepared from double-chained cationic surfactant, dihexadecyldimethylammonium bromide (DHDAB), and single-chained anionic surfactant, sodium dodecylsulfate (SDS). In order to manipulate the vesicular bilayer properties, cholesterol was used as an additive in the catanionic vesicle formation and the effects of cholesterol on the catanionic vesicle properties were evaluated. By using a forced formation process, DHDA-DS could form positively charged vesicles in aqueous phase with an average size of 132 nm and show good stability. With increased cholesterol content, it could suppress the phase transition behavior of DHDA-DS/cholesterol vesilces. Fluorescence polarization and infrared spectroscopy analyse showed that the addition of cholesterol into the DHDA-DS vesicles could have different effects on the vesicular membrane fluidity in different regions of the bilayer membranes. In addition, the zeta potential of the vesicles firstly increased with increasing dilution ratio and then decreased at high dilution ratio. Because of the positive charge on the vesicle surfaces, vesicle dispersions were stable when they were diluted. Human bronchial epithelial cells (BEAS-2B) were used to perform the cytotoxicity test with vesicle dispersions. In order to know exact cell viability, a small amount of DMSO solvent was added during the hydration process to reduce the vesicle aggregation. Moreover, the addition of cholesterol could increase the cell viability.

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