Study of Uptake Mechanisms of Halloysite Nanotubes in Different Cell Lines.
Study of Uptake Mechanisms of Halloysite Nanotubes in Different Cell Lines.
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DOI：
10.2147/ijn.s303816
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发表时间：
2021
影响因子：
8
通讯作者：
Di Blasi F
中科院分区：
文献类型：
--
作者：
Biddeci G;Spinelli G;Massaro M;Riela S;Bonaccorsi P;Barattucci A;Di Blasi F
硬铝石纳米管(HNTs)是一种化学式为Al2Si2O5(OH)4×nH2O的天然铝硅酸盐粘土，具有中空管状结构。由于其特殊的结构，HNTs可以作为药物载体系统发挥重要作用。目前，HNTs内化到活细胞的机制以及转运途径仍不清楚。因此，本研究以巨噬细胞系和非巨噬细胞系为模型，建立埃洛石纳米管内化的体外机制。HNT/CURBO杂化系统中，一个荧光探针(CURBO)被限制在HNT管腔内，作为一个模型来研究HNTs的运输途径机制。用MTS法检测HNT/CURBO在细胞模型上的细胞相容性。为了确定参与细胞摄取的内化途径，我们进行了各种内吞抑制研究，并使用荧光显微镜验证了细胞对纳米材料的内化。我们通过读取上清液在570 nm处的吸光度来评价HNT/CURBO与人红细胞(HRBC)接触的溶血效果。HNT/CURBO具有高度的生物相容性，没有明显的溶血作用。抑制试验的结果表明，在两种被研究的细胞系中，纳米管的内化过程是以能量依赖的方式发生的，尽管它们具有不同的特征。特别是，在非吞噬细胞中，参与了依赖于笼蛋白的和独立的内吞作用。在吞噬细胞中，除了吞噬和依赖于笼蛋白的内吞作用外，微管还参与埃洛伊特的细胞运输。在被细胞内化后，HNT/CURBO定位于细胞质区域，尤其是核周区域。了解HNTs的细胞转运途径有助于合理设计新型药物传递系统，对其在生物技术中的应用具有重要价值。
Halloysite nanotubes (HNTs) are a natural aluminosilicate clay with a chemical formula of Al2Si2O5(OH)4×nH2O and a hollow tubular structure. Due to their peculiar structure, HNTs can play an important role as a drug carrier system. Currently, the mechanism by which HNTs are internalized into living cells, and what is the transport pathway, is still unclear. Therefore, this study aimed at establishing the in vitro mechanism by which halloysite nanotubes could be internalized, using phagocytic and non-phagocytic cell lines as models. The HNT/CURBO hybrid system, where a fluorescent probe (CURBO) is confined in the HNT lumen, has been used as a model to study the transport pathway mechanisms of HNTs. The cytocompatibility of HNT/CURBO on cell lines model was investigated by MTS assay. In order to identify the internalization pathway involved in the cellular uptake, we performed various endocytosis-inhibiting studies, and we used fluorescence microscopy to verify the nanomaterial internalization by cells. We evaluated the haemolytic effect of HNT/CURBO placed in contact with human red blood cells (HRBCs), by reading the absorbance value of the supernatant at 570 nm. The HNT/CURBO is highly biocompatible and does not have an appreciable haemolytic effect. The results of the inhibition tests have shown that the internalization process of nanotubes occurs in an energy-dependent manner in both the investigated cell lines, although they have different characteristics. In particular, in non-phagocytic cells, clathrin-dependent and independent endocytosis are involved. In phagocytic cells, in addition to phagocytosis and clathrin-dependent endocytosis, microtubules also participate in the halloysite cellular trafficking. Upon internalization by cells, HNT/CURBO is localized in the cytoplasmic area, particularly in the perinuclear region. Understanding the cellular transport pathways of HNTs can help in the rational design of novel drug delivery systems and can be of great value for their applications in biotechnology.