開發癌細胞膜包覆的靛氰綠-喜樹鹼-全氟化奈米乳劑用於皮膚黑色素癌光化學治療;Development of Cancerous Membrane-Covered Indocyanine Green-Camptothecin-Loaded Perfluorinated Emulsions for Photochemotherapy of Melanoma

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題名:	開發癌細胞膜包覆的靛氰綠-喜樹鹼-全氟化奈米乳劑用於皮膚黑色素癌光化學治療;Development of Cancerous Membrane-Covered Indocyanine Green-Camptothecin-Loaded Perfluorinated Emulsions for Photochemotherapy of Melanoma
作者:	陳采昕;Chen, Cai-Sin
貢獻者:	生醫科學與工程學系
關鍵詞:	黑色素瘤;仿生奈米乳劑;靛氰綠;免疫逃逸;同源靶向;光熱療法;光動力療法;melanoma;biomimetic nanoemulsions;indocyanine green;immune escape;homologous targeting;photothermal therapy;photodynamic therapy
日期:	2023-07-25
上傳時間:	2024-09-19 15:19:59 (UTC+8)
出版者:	國立中央大學
摘要:	細胞膜包覆奈米粒子(或奈米乳劑)的技術正越來越多地被提及，而塗有細胞膜的奈米乳劑被預期會有一些優勢，首先利用細胞膜的外塗層作為天然靶點，可以解決外源性物質容易被免疫系統清除的問題，其次利用細胞膜偽裝奈米乳劑的目的是欺騙癌細胞，使其認為同樣是癌細胞，如此一來包裹在癌細胞膜中的奈米乳劑就可以聚集在癌細胞中，實現天然藥物靶向的功能。首先合成靛氰綠（Indocyanine green, ICG）-喜樹鹼（Camptothecin, CPT）-負載的全氟化奈米乳劑（Indocyanine green-camptothecin-loaded perfluorinated emulsions, ICPEs），結合化療和光療的協同治療，此外細胞膜是從小鼠黑色素瘤（B16F10）細胞中提取出來，再利用共擠壓的方式將ICPEs和癌細胞膜（Cancerous membranes, CMs）結合，最後形成外層癌細胞膜包覆內核奈米乳劑(Cancerous membrane-covered indocyanine green-camptothecin-loaded perfluorinated emulsions, CMICPEs)。 ICPEs的尺寸和電位測量值為230.43 ± 2.96 nm和-22.24 ± 1.48 mV，CMICPEs的尺寸和電位測量值為248.41 ± 8.33 nm和-32.78 ± 0.29 mV，結果顯示包覆細胞膜後尺寸增大。ICG和CPT在奈米乳劑中的包覆率分別為96.64 ± 2.81%和61.11 ± 11.86%。藥物釋放的評估試驗中可見CMICPEs在照射近紅外光（808 nm，6 W cm-2）下快速釋放藥物。ICPEs和CMICPEs在37℃下的ICG穩定性比游離的ICG提升了約1.5倍。在近紅外光（808 nm，6 W cm-2）照射下可以觀察到CMICPEs的光熱效應，並且CMICPEs的光動力效應優於游離的ICG。體外細胞毒性試驗結果可見CMICPEs照射NIR與相同藥量的free CPT相比更具細胞毒性，存活率分別為6.06%和64.70%，有顯著差異，在體內的腫瘤療效試驗中，證實CMICPEs有抑制腫瘤的效果，並且在體內螢光成像試驗中，觀察到CMICPEs具有良好腫瘤滯留效果與腫瘤的靶向能力。從以上的實驗評估中，我們可以期望這項技術的發展，並且預期它作為一種新興的癌症治療材料的潛力。;Cell membrane-encapsulated nanoparticle (or nanoemulsion) technology has attracted significant attention in recent times. Nanoemulsions coated with cell membranes exhibit some advantages. First, using the outer coating of the cell membrane as a natural target may solve the problem of exogenous material being targeted and cleared by the immune system. Second, the nanoemulsions disguised by cell membranes can be misrecognized by cancer cells as their own; hence, nanoemulsions encased in cancer-cell membranes can reach other cancer cells and achieve natural drug targeting. Herein, indocyanine green (ICG)-camptothecin (CPT)-loaded perfluorinated emulsions (ICPEs) were synthesized. Cell membranes were extracted from melanoma (B16F10), and coextrusion was employed to combine the ICPEs with cancerous membranes (CMs). Finally, CM-covered ICPEs (CMICPEs) were formed. The size and potential measurements of the ICPEs and CMICPEs were 230.43±2.96 nm and –22.24±1.48 mV and 248.41±8.33 nm and –32.78±0.29 mV, respectively. The sizes increased after coating the cell membrane. The encapsulation rates of the ICG and CPT in the nanoemulsions were 96.64±2.81 and 61.11±11.86%, respectively. CMICPEs can achieve rapid drug release under near-infrared irradiation (NIR, 808 nm, 6 W cm-2). The ICG stability of the ICPEs and CMICPEs at 37 °C increased by approximately 1.5 times the stability exhibited by free ICG. The photothermal effect of the CMICPEs was observed under NIR irradiation (808 nm, 6 W cm-2). The photodynamic effect of the CMICPEs was determined to be better than that of the free ICG. In vitro cytotoxicity test results indicated that the CMICPEs under NIR were more cytotoxic than free CPT at the same dose, with significant differences in their survival rates (6.06 and 64.70%, respectively). In an in vivo tumor therapy test, CMICPEs were shown to have tumor suppressive effects, and good tumor retention and tumor targeting ability were observed in an in vivo fluorescence imaging test. We expect this technology to be further developed using these experimental results, and we anticipate its potential as an emerging material for cancer therapy.
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