Vicente-Zurdo David, Gómez-Gómez Beatriz, Romero-Sánchez Iván, Rosales-Conrado Noelia, León-González María Eugenia, Madrid Yolanda
Analytical Chemistry Department, Faculty of Chemistry Sciences, Complutense University of Madrid, E-28040, Madrid, Spain.
Analytical Chemistry Department, Faculty of Chemistry Sciences, Complutense University of Madrid, E-28040, Madrid, Spain.
Anal Chim Acta. 2023 Apr 8;1249:340949. doi: 10.1016/j.aca.2023.340949. Epub 2023 Feb 6.
Alzheimer's disease (AD) is the most prevalent neurodegenerative disease, representing 80% of the total dementia cases. The "amyloid cascade hypothesis" stablishes that the aggregation of the beta-amyloid protein (Aβ) is the first event that subsequently triggers AD development. Selenium nanoparticles stabilized with chitosan (Ch-SeNPs) have demonstrated excellent anti-amyloidogenic properties in previous works, leading to an improvement of AD aetiology. Here, the in vitro effect of selenium species in AD model cell line has been study to obtain a better assessment of their effects in AD treatment. For this purpose, mouse neuroblastoma (Neuro-2a) and human neuroblastoma (SH-SY5Y) cell lines were used. Cytotoxicity of selenium species, such as selenomethionine (SeMet), Se-methylselenocysteine (MeSeCys) and Ch-SeNPs, has been determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry methods. Intracellular localisation of Ch-SeNPs, and their pathway through SH-SY5Y cell line, have been evaluated by transmission electron microscopy (TEM). The uptake and accumulation of selenium species by both neuroblastoma cell lines have been quantified at single cell level by single cell- Inductively Coupled Plasma with Mass Spectrometry detection (SC-ICP-MS), with a previous optimisation of transport efficiency using gold nanoparticles (AuNPs) ((69 ± 3) %) and 2.5 mm calibration beads ((92 ± 8) %). Results showed that Ch-SeNPs would be more readily accumulated by both cell lines than organic species being accumulation ranges between 1.2 and 89.5 fg Se cell for Neuro-2a and 3.1-129.8 fg Se cell for SH-SY5Y exposed to 250 μM Ch-SeNPs. Data obtained were statistically treated using chemometric tools. These results provide an important insight into the interaction of Ch-SeNPs with neuronal cells, which could support their potential use in AD treatment.
阿尔茨海默病（AD）是最常见的神经退行性疾病，占所有痴呆病例的80%。“淀粉样蛋白级联假说”认为，β-淀粉样蛋白（Aβ）的聚集是随后引发AD发展的首要事件。壳聚糖稳定的硒纳米颗粒（Ch-SeNPs）在先前的研究中已显示出优异的抗淀粉样蛋白生成特性，有助于改善AD病因。在此，对AD模型细胞系中硒物种的体外效应进行了研究，以更好地评估它们在AD治疗中的作用。为此，使用了小鼠神经母细胞瘤（Neuro-2a）和人神经母细胞瘤（SH-SY5Y）细胞系。通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）和流式细胞术方法测定了硒代蛋氨酸（SeMet）、硒甲基硒代半胱氨酸（MeSeCys）和Ch-SeNPs等硒物种的细胞毒性。通过透射电子显微镜（TEM）评估了Ch-SeNPs在细胞内的定位及其通过SH-SY5Y细胞系的途径。通过单细胞电感耦合等离子体质谱检测（SC-ICP-MS）在单细胞水平上对两种神经母细胞瘤细胞系对硒物种的摄取和积累进行了定量，此前使用金纳米颗粒（AuNPs）（(69 ± 3)%）和2.5 mm校准珠（(92 ± 8)%）对转运效率进行了优化。结果表明，与有机物种相比，两种细胞系对Ch-SeNPs摄取更快，暴露于250 μM Ch-SeNPs时，Neuro-2a细胞中Ch-SeNPs的积累范围为1.2至89.5 fg Se/细胞，SH-SY5Y细胞中为3.1至129.8 fg Se/细胞。使用化学计量工具对获得的数据进行了统计学处理。这些结果为Ch-SeNPs与神经元细胞的相互作用提供了重要见解，这可能支持它们在AD治疗中的潜在应用。
