光熱治療於 Cal 27 細胞之測試

確定本研究所合成之奈米複合材料均無細胞毒性之疑慮後，將利用金奈米粒 子之光熱轉換特性，應用於光熱治療，藉由光熱轉換所產生之熱能導致細胞死亡。

光熱治療所選用之細胞株為對二氧化矽修飾之奈米複合材料具較良好細胞存活 率之口腔癌細胞 Cal 27。實驗過程如圖 3-22 所示，將濃度為 100 μg/mL 奈米複 合材料餵食於 Cal 27 細胞後，以功率為 1.7 W/cm²之 980 nm 紅外光雷射照射一 分鐘後，加入台盼藍(trypan blue)進行染色，破碎之細胞將殘留 trypan blue 染劑 於其細胞質內，因此經光學顯微鏡觀察若具深藍色之區域即為細胞死亡之現象。

圖 3-22 光熱治療過程示意圖。

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光熱治療之結果如圖 3-23 與圖 3-24 所示，圖 3-23 中之四組對照組分別為 僅具 Cal 27 細胞、加入二氧化矽修飾之上轉換奈米粒子(UCNP@SiO2)、加入球 狀金奈米粒子(Au NP)與加入棒狀金奈米粒子(Au NR)之四種組別，雷射照射後 並以台盼藍進行染色後於顯微鏡下觀察，可發現四組皆無細胞死亡之現象。

圖 3-23 光熱治療實驗於口腔癌細胞 Cal 27 之對照組結果。

圖 3-24 為 修 飾 球 狀 金 奈 米 粒 子 (UCNP@SiO2-AuNP) 及 棒 狀 金 奈 米 粒 子 (UCNP@SiO2-AuNR) 之組別，分別加入低濃度與高濃度之金奈米粒子，可發現 UCNP@SiO2-AuNP 組別於雷射照射處具少數細胞死亡現象而添加較高濃度之球 狀金奈米粒子後具稍大範圍細胞死亡之現象，此外 UCNP@SiO2-AuNR 之組別則 具相當明顯之細胞死亡現象，而添加高濃度之棒狀金奈米粒子後細胞呈現大範圍 死亡之現象。由上述之結果可發現光熱治療之效果隨修飾較多金奈米粒子而提高，

另一部分則為球狀金奈米粒子與棒狀金奈米粒子之差異，造成如此明顯差異之原 因推測為棒狀金奈米粒子具較高之消光係數(extinction coefficient)，可吸收較多 光能並轉換為熱能，藉以提供較高效率之光熱治療效果。

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圖 3-24 光熱治療實驗於口腔癌細胞 Cal 27 之結果。

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第四章 結論

(1) 本研究所製備之二氧化矽修飾之上轉換奈米粒子於水溶液中以單顆分散且 無聚集之現象發生，無論修飾球狀金奈米粒子或棒狀金奈米粒子於其表面後 亦呈現相當均勻之分散性，並於細胞毒性測試中證實本研究所合成之奈米複 合材料具低生物毒性，對於往後於體內(in vivo)之研究具相當大潛力。

(2) 本研究所製備之多功能奈米複合材料可透過 980 nm 之雷射激發，同時達到 細胞標定與光熱治療之效果，上轉換奈米粒子之特性放光可透過共軛聚焦顯 微鏡驗證其於細胞標定之能力。另一方面，透過表面修飾金奈米粒子，利用 上轉換奈米粒子之特性放光與表面電漿共振峰相重疊之特性，將光能轉換為 熱能以提供光熱治療之熱能來源致使細胞死亡。而本研究亦透過合成不同形 貌之金奈米粒子以探討其光熱轉換之特性，於口腔癌細胞 cal 27 之光熱治療 實驗中可發現，棒狀金奈米粒子之光熱治療效果較優於球狀金奈米粒子，且 隨棒狀金奈米粒子比例增加，光熱治療之效果愈趨顯著。

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