Folic-Acid-Functionalized Graphene Oxide Nanocarrier: Synthetic Approaches, Characterization, Drug Delivery Study, and Antitumor Screening
Folic-Acid-Functionalized Graphene Oxide Nanocarrier: Synthetic Approaches, Characterization, Drug Delivery Study, and Antitumor Screening
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DOI：
10.1021/acsanm.7b00324
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发表时间：
2018-02-01
影响因子：
5.9
通讯作者：
Alves, Oswaldo Luiz
中科院分区：
文献类型：
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作者：
de Sousa, Marcelo;Visani de Luna, Luis Augusto;Alves, Oswaldo Luiz
在这项工作中，我们开发并筛选了基于氧化石墨烯（GO）和叶酸（FA）的纳米载体用于递送化疗药物的潜在抗肿瘤活性。通过石墨剥离工艺合成GO。将FA连接至PEG（4，7，10-三氧杂-1，13-十三烷二胺）以形成FA PEG，然后偶联至GO表面。喜树碱（CPT）被进一步吸附在GO上用作递送研究中的药物模型。中间体FA-PEG分子的合成以及与GO的偶联以形成GO-FA纳米载体通过基础和最先进的表征技术来证实，包括红外（FTIR）光谱、热重分析（TGA）、电喷雾电离（ESI）质谱、透射电子显微镜（TEM）、透射电子显微镜和魔角旋转碳-13核磁共振（CP/MAS C-13 NMR）光谱。FTIR光谱显示在用FA-PEG官能化GO后羧基的信号强度显著降低。GO-FA的TGA显示约20%的官能团来自FA-PEG。GO FA显示高CPT负载能力（37.8%）。体外研究证实了超过200小时的药物释放延长。与生理pH（7.4）相比，酸性pH（5.0）减缓CPT从纳米载体的释放。针对两种广泛研究的临床前细胞模型研究了GO FA和GO FA + CPT的毒性筛选：J774，一种具有巨噬细胞表型和高增殖率的肿瘤细胞;和HepG 2，一种从人肝细胞癌获得的具有叶酸转运蛋白的肿瘤细胞。未载药的GO-FA纳米载体的毒性依赖于细胞类型，并且对J774没有毒性，但对HepG 2有高毒性。FA在载有CPT的纳米载体中的存在对于在两种肿瘤细胞系中实现凋亡至关重要。此外，共聚焦显微镜显示的FITC标记的GO-FA的粘附和内化的肿瘤细胞系。
In this work, we developed and screened the potential antitumor activity of a nanocarrier based on graphene oxide (GO) and folic acid (FA) for the delivery of chemotherapy drugs. GO was synthesized by the graphite exfoliation process. FA was linked to PEG (4,7,10-trioxa-1,13-tridecanediamine) to form FA PEG, followed by coupling to the GO surface. Camptothecin (CPT) was further adsorbed on GO for use as a drug model in the delivery study. The synthesis of the intermediate FA-PEG molecule and coupling to GO for the formation of the GO-FA nanocarrier were confirmed by basic and state-of-the-art characterization techniques, including infrared (FTIR) spectroscopy, thermogravimetric analysis (TGA), electrospray ionization (ESI) mass spectrometry, transmission electron microscopy (TEM), and magic-angle spinning carbon-13 nuclear magnetic resonance (CP/MAS C-13 NMR) spectroscopy. FTIR spectroscopy showed a significant reduction in the signal intensity of the carboxylic groups after the functionalization of GO with FA-PEG. TGA of GO-FA revealed that approximately 20% of the functional groups were from FA-PEG. GO FA indicated a high CPT loading capacity (37.8%). In vitro studies confirmed prolonged drug release over 200 h. Acidic pH (5.0) slowed the release of CPT from the nanocarrier compared to that at physiological pH (7.4). The toxicity screening of GO FA and GO FA + CPT was investigated for two widely studied preclinical cell models: J774, a tumor cell with macrophage phenotype and high proliferation rate; and HepG2, a tumor cell obtained from human hepatocellular carcinoma with folate transporters. The toxicity of the GO-FA nanocarrier without drug loading was dependent on the cell type and presented no toxicity to J774 but high toxicity to HepG2. The presence of FA in the nanocarrier loaded with CPT was crucial to achieve apoptosis in both tumor cell lines. In addition, confocal microscopy revealed both the adhesion and internalization of the FITC-labeled GO-FA by the tumor cell lines.