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    题名: 製備包覆靛氰綠之聚乳酸甘醇酸標靶奈米粒子用於乳癌光熱暨光動治療之研究;Fabrication of the Targeting Indocyanine Green-Encapsulated PLGA Nanoparticles for Phototherapy of Breast Cancer.
    作者: 賴允涵;Lai,Yun-han
    贡献者: 生物醫學工程研究所
    关键词: PLGA奈米粒子;靛氰綠;光治療;乳癌
    日期: 2015-08-26
    上传时间: 2015-09-23 10:56:27 (UTC+8)
    出版者: 國立中央大學
    摘要: 本研究我們使用微乳液法合成出包覆靛氰綠(Indocyanine Green;ICG)之聚乳酸甘醇酸(Poly(Lactic-co-Glycolic Acid);PLGA)的標靶奈米粒子(HER-2 Targeted ICG-Encapsulated PLGA Nanoparticles;HIPNPs),其中我們利用交聯反應將聚乙二醇與乳癌細胞表面受器抗體Human Epidermal Growth Factor Receptor 2 (HER-2)依序接枝在奈米粒子表面,以增進奈米粒子的生物相容性與對乳癌細胞專一標定的功能,並利用傅立葉轉換紅外線光譜儀分析粒子表面官能基與結合螢光抗體的方式來證明聚乙二醇跟乳癌抗體的成功接枝。經過儀器分析奈米粒子之平均粒徑與表面電位分別為302  1.8 nm和-15  0.15 mV;而ICG的包覆率約為70%。再來我們利用激發波長808 nm搭配強度為6 W/cm2的近紅外光雷射照射HIPNPs奈米溶液,結果顯示在含有25 μM的HIPNPs溶液中,照射2分鐘內溫度即可到達70 oC的高溫且該溫度可持續5分鐘,同時在相同雷射條件刺激下,我們測量到90單位的單態氧的釋放。接著我們將不同濃度的HIPNPs溶液與MDA-MB-453細胞一同培養24小時後分析得到細胞的存活率為90 %,以此我們可確認HIPNPs不具生物毒性。另外,藉由MDA-MB-453 (HER-2 Positive)的螢光表現量遠大於MCF-7 (HER-2 Negative)的結果,我們可確認HIPNPs對HER-2+細胞的專一性。最後,我們使用激發波長808 nm且強度為6 W/cm2的近紅外光雷射照射內吞HIPNPs的MDA-MB-453乳癌細胞5分鐘,經由計算得知細胞的存活率僅存6 %,證明了HIPNPs可有效進行光治療使乳癌細胞死亡,並有望進而發展成為一種治療癌症的材料。;The biodegradable indocyanine green (ICG)-loaded poly(lactic-co-glycolic acid) nanoparticles (IPNPs) have been successfully established by using solvent evaporation method. Furthermore we conjugated the functional molecules like Polyethylene glycol (PEG) and human epithelial receptor-2 (HER-2) antibody onto the IPNPs surface by utiliznig the EDC/NHS crosslinking method to fabricate the HER-2 targeted ICG-loaded PLGA nanoparticles (HIPNPs) for phototherapy of breast cancer cells. The ICG encapsulation efficiency was about 70 % determined from the UV-Vis spectrophotometry. The mean size and surface charge of the HIPNPs were 302 ± 1.8 nm and -15 ± 0.15mV through dynamic light scattering and zeta potential analyzer, respectively. Based on the analysis of UV-Vis spectrophotometric absorbances, the encapsulated ICG modestly disintegrated 18% and 63% while ICG freely distributed in water dramatically degraded 60% and 95% at 4 and 37℃ for 48 h, respectively, manifesting the ICG molecules in HIPNPs can be protected by PLGA matrix. In addition, the bulk temperature dramatically increased 45℃ and the singlet oxygen released in the presence of HIPNPs (ICG concentration = 25 μM) under 808 nm-laser exposure with intensity of 6 W/cm2 for 5 min has successfully proved the photothermal and photodynamic function of ICG. Finally through the cellular assay using MCF-7 (HER-2 negative), and MDA-MB-453 (HER-2 positive), we found HIPNPs can specifically target to the HER-2 protein and didn’t have biotoxicity. Even, the nmber of survival MDA-MB-453 was significantly low by treating HIPNPs with identical laser exposure, identifying the effectiveness of phototherapy of HIPNPs. We demonstrated the developed HIPNPs enable to provide the theranostic efficacy for breast cancer cells
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