Institute of Chemistry, University of Silesia in Katowice, Katowice, 40-006, Poland.
Silesian Center for Education and Interdisciplinary Research, 75 Pulku Piechoty 1a, 41-500 Chorzow, Poland.
ACS Biomater Sci Eng. 2022 Aug 8;8(8):3450-3462. doi: 10.1021/acsbiomaterials.2c00542. Epub 2022 Jul 20.
Cellular localization of carbon nanomaterials in cancer cells is essential information for better understanding their interaction with biological targets and a crucial factor for further evaluating their biological properties as nanovehicles or nanotherapeutics. Recently, increasing efforts to develop promising fullerene nanotherapeutics for cancer nanotechnology have been made. However, the main challenge regarding studying their cellular effects is the lack of effective methods for their visualization and determining their cellular fate due to the limited fluorescence of buckyball scaffolds. Herein, we developed a method for cellular localization of nonfluorescent and water-soluble fullerene nanomaterials using the click chemistry approach. First, we synthesized a triple-bonded fullerene probe (TBCser), which was further used as a starting material for 1,3-dipolar cycloaddition using 3-azido-7-hydroxycoumarin and sulfo-cyanine5 azide fluorophores to create fluorescent fullerene triazoles. In this work, we characterized the structurally triple-bonded [60]fullerene derivative and confirmed its high symmetry () and the successful formation of fullerene triazoles by spectroscopic techniques (i.e., ultraviolet-visible, fluorescence, and Fourier transform infrared spectroscopies) and mass spectrometry. The created fluorescent fullerene triazoles were successfully localized in the MCF-7 breast cancer cell line using fluorescent microscopy. Overall, our findings demonstrate that TBCser localizes in the lysosomes of MCF-7 cells, with only a small affinity to mitochondria.
碳纳米材料在癌细胞中的细胞定位对于更好地了解它们与生物靶标的相互作用是必不可少的信息,也是进一步评估它们作为纳米载体或纳米治疗剂的生物特性的关键因素。最近,人们越来越努力地开发有前途的富勒烯纳米治疗剂用于癌症纳米技术。然而,由于富勒烯支架的荧光有限,研究它们的细胞效应的主要挑战是缺乏有效的可视化方法和确定它们的细胞命运的方法。在这里,我们开发了一种使用点击化学方法对非荧光和水溶性富勒烯纳米材料进行细胞定位的方法。首先,我们合成了一个三键结合的富勒烯探针(TBCser),它进一步用作使用 3-叠氮-7-羟基香豆素和磺基花青 5 叠氮荧光团进行 1,3-偶极环加成的起始材料,以创建荧光富勒烯三唑。在这项工作中,我们对结构上三键结合的 [60]富勒烯衍生物进行了表征,并通过光谱技术(即紫外-可见、荧光和傅里叶变换红外光谱)和质谱证实了其高对称性()和富勒烯三唑的成功形成。创建的荧光富勒烯三唑成功地使用荧光显微镜定位于 MCF-7 乳腺癌细胞系中。总的来说,我们的研究结果表明,TBCser 定位于 MCF-7 细胞的溶酶体中,与线粒体的亲和力很小。
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