Hifni Badriah, Khan Mona, Devereux Stephen J, Byrne Maria H, Quinn Susan J, Simpson Jeremy C
School of Chemistry, University College Dublin, Belfield, Dublin 4 D04 N2E5, Ireland.
School of Biology & Environmental Science, University College Dublin, Belfield, Dublin 4 D04 N2E5, Ireland.
ACS Appl Bio Mater. 2020 Oct 19;3(10):6790-6801. doi: 10.1021/acsabm.0c00748. Epub 2020 Sep 17.
The high surface area, facile functionalization, and biocompatibility of carbon nanohorns (CNHs) make them attractive for many applications, including drug delivery. The cellular destination of nanomaterials dictates both the therapeutic application and the potential toxicity. Identifying the uptake mechanism is challenging as several endocytic pathways have been identified that facilitate cellular entry. Here, the cellular uptake of fluorescently labeled CNHs was assessed by utilizing quantitative cell-based assays to determine the factors influencing how internalization occurs and the destinations they reach in HeLa cells. Cell viability assays suggest that about 80% of the cells remained viable even at the highest concentration of 20 μg/mL exposure to CNHs. Uptake studies revealed that when pulse-chase conditions were applied, CNHs were seen to be localized both at the cell periphery and in a juxtanuclear pattern inside HeLa cells, in the latter case colocalizing with the lysosomal marker LAMP1. RNA interference studies, using a panel of RNA tools to individually deplete key molecules associated with the endocytic machinery, failed to block the internalization of CNHs into cells, suggesting that multiple mechanisms of endocytosis are used by this particle type.
碳纳米角（CNHs）具有高比表面积、易于功能化和生物相容性，这使其在包括药物递送在内的许多应用中具有吸引力。纳米材料的细胞归宿决定了其治疗应用和潜在毒性。由于已确定多种促进细胞摄取的内吞途径，因此确定摄取机制具有挑战性。在此，通过利用基于细胞的定量分析来评估荧光标记的CNHs的细胞摄取，以确定影响内化发生方式及其在HeLa细胞中到达位置的因素。细胞活力分析表明，即使在最高浓度20μg/mL的CNHs暴露下，仍有约80%的细胞保持活力。摄取研究表明，当采用脉冲追踪条件时，在HeLa细胞中可观察到CNHs定位于细胞周边以及细胞核旁，在后一种情况下与溶酶体标记物LAMP1共定位。使用一组RNA工具分别耗尽与内吞机制相关的关键分子的RNA干扰研究未能阻止CNHs内化进入细胞，这表明这种颗粒类型利用了多种内吞机制。
