López-Martín Raúl, Aranda-Sobrino Nieves, De Enciso-Campos Nerea, Sánchez Elena H, Castañeda-Peñalvo Gregorio, Lee Su Seong, Binns Chris, Ballesteros-Yáñez Inmaculada, De Toro Jose A, Castillo-Sarmiento Carlos A
Departamento de Física Aplicada, Instituto Regional de Investigación Científica Aplicada (IRICA), Universidad de Castilla-La Mancha, Ciudad Real 13071, Spain.
Department of Inorganic and Organic Chemistry and Biochemistry, School of Medicine, University of Castilla-La Mancha, Ciudad Real 13071, Spain.
R Soc Open Sci. 2024 Jun 19;11(6):231839. doi: 10.1098/rsos.231839. eCollection 2024 Jun.
Nanoparticle uptake by cells is a key parameter in their performance in biomedical applications. However, the use of quantitative, non-destructive techniques to obtain the amount of nanoparticles internalized by cells is still uncommon. We have studied the cellular uptake and the toxicity of core-shell maghemite-silica magnetic nanoparticles (MNPs), with a core diameter of 9 nm and a shell thickness of 3 nm. The internalization of the nanoparticles by mouse neuroblastoma 2a cells was evaluated by sensitive and non-destructive Superconducting Quantum Interference Device (SQUID) magnetometry and corroborated by graphite furnace atomic absorption spectroscopy. We were thus able to study the toxicity of the nanoparticles for well-quantified MNP uptake in terms of nanoparticle density within the cell. No significant variation in cell viability or growth rate was detected for any tested exposure. Yet, an increase in both the amount of mitochondrial superoxide and in the lysosomal activity was detected for the highest concentration (100 μg ml) and incubation time (24 h), suggesting the onset of a disruption in ROS homeostasis, which may lead to an impairment in antioxidant responses. Our results validate SQUID magnetometry as a sensitive technique to quantify MNP uptake and demonstrate the non-toxic nature of these core-shell MNPs under our culture conditions.
细胞对纳米颗粒的摄取是其在生物医学应用中性能的一个关键参数。然而,使用定量、非破坏性技术来获取细胞内化纳米颗粒的量仍然并不常见。我们研究了核壳结构的磁赤铁矿-二氧化硅磁性纳米颗粒(MNPs)的细胞摄取及其毒性,其核心直径为9纳米,壳层厚度为3纳米。通过灵敏且非破坏性的超导量子干涉装置(SQUID)磁力测定法评估了小鼠神经母细胞瘤2a细胞对纳米颗粒的内化情况,并通过石墨炉原子吸收光谱法进行了验证。因此,我们能够根据细胞内纳米颗粒的密度,研究纳米颗粒对定量摄取的纳米颗粒的毒性。对于任何测试的暴露情况,均未检测到细胞活力或生长速率有显著变化。然而,在最高浓度(100μg/ml)和孵育时间(24小时)下,检测到线粒体超氧化物的量和溶酶体活性均增加,这表明活性氧(ROS)稳态开始受到破坏,这可能导致抗氧化反应受损。我们的结果验证了SQUID磁力测定法作为一种量化纳米颗粒摄取的灵敏技术,并证明了在我们的培养条件下这些核壳结构纳米颗粒的无毒性质。