具葉酸受器標靶特性之奈米金桿於光熱與光動力治療探討

The Study of Gold Nanorods for Folate Receptor Targeted Photothermal and Photodynamic Therapy

郭書妤(Shu-Yu Guo)

指導教授 ： 李文婷

中原大學/工學院/生物醫學工程研究所/碩士(2015年)

https://doi.org/10.6840/CYCU.2015.00156

摘要

本研究合成水相的奈米金桿 (Gold nanorods, GNRs) 作為光感藥物靛花青素 (Indocyanine green, ICG) 與葉酸 (Folic acid, FA) 的載體，並探討其在人類表皮樣癌-表皮細胞生長因子受體 (Epithelial growth factor receptor, EGFR) 過表達的 A 431 細胞以及葉酸受體 (Folate receptor, FR) 過表達的KB 細胞之光熱 (Photothermal Therapy, PTT) 與光動力治療 (Photodynamic Therapy, PDT) 應用。首先製備出 GNR-PEG-ICG、GNR-PEG-FA 與 GNR-PEG-ICG-FA 三種藥物，並利用紫外光/可見光光譜、傅立葉轉換紅外線光譜及螢光光譜確認 PEG-ICG 與 PEG-FA 已成功接枝至 GNRs 上。此外，由表面電位及膠體電泳結果可推測 GNRs 因表面帶有溴化十六烷基三甲銨 (Cetyltrimethylammonium bromide, CTAB)，所以呈現正電；而 GNR-PEG-ICG 表面則帶有負電荷；GNR- PEG-FA 與 GNR-PEG-ICG-FA 表面電位皆為電中性。ICG、GNRs、GNR-PEG- ICG 及 GNR-PEG-ICG-FA 皆有光熱效應產生，而 ICG 及 GNRs 的溫度變化較大。體外實驗發現 KB 細胞僅在 4 小時就達到對 GNR-PEG-ICG-FA 的最大攝取量；添加 FA 可顯著抑制 KB 細胞對 GNR-PEG-ICG-FA 的攝入，證實 GNR-PEG-ICG-FA 對 KB 細胞有標靶的功用。以波長 780 nm、能量 50 J/cm2 光源搭配不同藥物對細胞進行 PDT-PTT，發現 GNR-PEG-ICG 對 A431 細胞的半致死劑量 (LD50) 為 76.9±20.2 nM，而 GNR-PEG-ICG-FA對 A431 細胞的 LD50 為 254.1±12.0 nM；GNR-PEG-ICG 對 KB 細胞的 LD50 為 100.0± 14.0 nM， 而 GNR-PEG-ICG-FA 對 KB 細胞的 LD50 則下降為 58.6±0.7 nM。綜上所述，本研究成功地開發出具有 FR 專一性的 GNR-PEG-ICG-FA，並驗證其對 KB 細胞的 PDT-PTT 療效。

關鍵字

光熱治療 ； 光動力治療 ； 葉酸 ； 靛花青素 ； 奈米金桿

並列摘要

The study was to investigate the application of water-soluble gold nanorods (GNRs) as indocyanine green (ICG) and folic acid (FA) delivering vectors for photodynamic therapy (PDT) and photothermal therapy (PTT) against human epidermoid carcinoma, epithelial growth factor receptor (EGFR) overexpressed A 431 cells and folate receptor (FR) overexpressed KB cells. Ultraviolet-visible spectroscopy, Fourier transformed infrared spectroscopy and fluorescence spectroscopy showed that PEG-ICG and PEG-FA were successfully conjugated to GNRs to prepare GNR-PEG-ICG, GNR-PEG-FA and GNR-PEG-ICG-FA. Zeta potential analyses and agarose gel electrophoresis revealed that GNRs carried positive charge due to cetyltrimethylammonium bromide (CTAB) attached on their surface. GNR-PEG-ICG carried negative charge, whereas GNR-PEG-FA and GNR-PEG-ICG-FA possessed electroneutrality. ICG, GNRs, GNR-PEG-ICG and GNR-PEG-ICG-FA all showed photo-thermal effect of which higher temperature elevation was observed in ICG and GNRs. In vitro study found that the accumulation of GNR-PEG-ICG-FA in KB cells achieved maximum after 4 hours incubation. The targeting ability of GNR-PEG-ICG-FA to KB cells was demonstrated by observing the uptake inhibition of GNR-PEG-ICG-FA with the addi- tion of free FA. PDT-PTT effect was evaluated with light at wavelength of 780 nm and energy density of 50 J/cm2 in combination of different drugs. Half lethal dose (LD50) of GNR-PEG-ICG on A431 cells was 76.9 ±20.2 nM, whereas LD50 of GNR-PEG- ICG-FA was 254.1±12.0 nM. LD50 of GNR-PEG-ICG on KB cells was 100.0±14.0 nM, whereas LD50 of GNR-PEG-ICG-FA decreased to 58.6±0.7 nM. In summary, GNR- PEG-ICG-FA with FR specificity was successfully developed and its PDT-PTT effect on KB cells was demonstrated.

並列關鍵字

gold nanorod ； folic acid ； photodynamic therapy ； photothermal therapy ； indocyanine green

參考文獻

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