Zimmermann Kristen A, Inglefield David L, Zhang Jianfei, Dorn Harry C, Long Timothy E, Rylander Christopher G, Rylander M Nichole
School of Biomedical Engineering and Sciences, Virginia Polytechnic Institute and State University, ICTAS Building, Stanger Street, Blacksburg, VA 24061, USA.
Department of Chemistry, Virginia Polytechnic Institute and State University, ICTAS II Building, Washington Street, Blacksburg, VA 24061, USA.
J Nanopart Res. 2014 Jan;16(1). doi: 10.1007/s11051-013-2078-3. Epub 2013 Dec 4.
Single-walled carbon nanohorns (SWNHs) have great potential to enhance thermal and chemotherapeutic drug efficiencies for cancer therapies. Despite their diverse capabilities, minimal research has been conducted so far to study nanoparticle intracellular transport, which is an important step in designing efficient therapies. SWNHs, like many other carbon nanomaterials, do not have inherent fluorescence properties making intracellular transport information difficult to obtain. The goals of this project were to (1) develop a simple reaction scheme to decorate the exohedral surface of SWNHs with fluorescent quantum dots (QDs) and improve conjugate stability, and (2) evaluate SWNH-QD conjugate cellular uptake kinetics and localization in various cancer cell lines of differing origins and morphologies. In this study, SWNHs were conjugated to CdSe/ZnS core/shell QDs using a unique approach to carbodiimide chemistry. Transmission electron microscopy and electron dispersive spectroscopy verified the conjugation of SWNHs and QDs. Cellular uptake kinetics and efficiency were characterized in three malignant cell lines: U-87 MG (glioblastoma), MDA-MB-231 (breast cancer), and AY-27 (bladder transitional cell carcinoma) using flow cytometry. Cellular distribution was verified by confocal microscopy, and cytotoxicity was also evaluated using an alamarBlue assay. Results indicate that cellular uptake kinetics and efficiency are highly dependent on cell type, highlighting the significance of studying nanoparticle transport at the cellular level. Nanoparticle intracellular transport investigations may provide information to optimize treatment parameters (e.g., SWNH concentration, treatment time, etc.) depending on tumor etiology.
单壁碳纳米角(SWNHs)在提高癌症治疗的热疗和化疗药物效率方面具有巨大潜力。尽管它们具有多种功能,但迄今为止,对于纳米颗粒细胞内运输的研究却很少,而这是设计有效治疗方法的重要一步。与许多其他碳纳米材料一样,SWNHs没有固有的荧光特性,因此难以获得细胞内运输信息。本项目的目标是:(1)开发一种简单的反应方案,用荧光量子点(QDs)修饰SWNHs的外表面并提高共轭稳定性;(2)评估SWNH-QD共轭物在不同来源和形态的各种癌细胞系中的细胞摄取动力学和定位。在本研究中,采用一种独特的碳二亚胺化学方法将SWNHs与CdSe/ZnS核/壳量子点共轭。透射电子显微镜和电子色散光谱证实了SWNHs和量子点的共轭。使用流式细胞术在三种恶性细胞系中表征细胞摄取动力学和效率:U-87 MG(胶质母细胞瘤)、MDA-MB-231(乳腺癌)和AY-27(膀胱移行细胞癌)。通过共聚焦显微镜验证细胞分布,并使用alamarBlue测定法评估细胞毒性。结果表明,细胞摄取动力学和效率高度依赖于细胞类型,突出了在细胞水平研究纳米颗粒运输的重要性。纳米颗粒细胞内运输研究可能会提供信息,以便根据肿瘤病因优化治疗参数(例如,SWNH浓度、治疗时间等)。