Leite Paulo Emílio Corrêa, Pereira Mariana Rodrigues, do Nascimento Santos Carlos Antonio, Campos Andrea Porto Carreiro, Esteves Ticiana Mota, Granjeiro José Mauro
Divisão de Biologia Celular e Bioengenharia, Diretoria de Metrologia Aplicada as Ciências da Vida (DIMAV), Brazil.
Instituto de Biologia, Universidade Federal Fluminense, Niterói, RJ, Brazil.
Toxicol In Vitro. 2015 Aug;29(5):819-27. doi: 10.1016/j.tiv.2015.02.010. Epub 2015 Mar 16.
Gold nanoparticles (AuNP) have been widely used for many applications, including as biological carriers. A better understanding concerning AuNP safety on muscle cells is crucial, since it could be a potential tool in the nanomedicine field. Here, we describe the impact of polyethylene glycol-coated gold nanoparticles (PEG-AuNP) interaction with differentiated skeletal muscle C2C12 cells on cell viability, mitochondria function, cell signaling related to survival, cytokine levels and susceptibility to apoptosis. Intracellular localization of 4.5 nm PEG-AuNP diameter size was evidenced by STEM-in-SEM in myotube cells. Methods for cytotoxicity analysis showed that PEG-AuNP did not affect cell viability, but intracellular ATP levels and mitochondrial membrane potential increased. Phosphorylation of ERK was not altered but p-AKT levels reduced (p<0.01). Pre-treatment of cells with PEG-AuNP followed by staurosporine induction increased the caspases-3/7 activity. Indeed, cytokines analysis revealed a sharp increase of IFN-γ and TGF-β1 levels after PEG-AuNP treatment, suggesting that inflammatory and fibrotic phenotypes process were activated. These data demonstrate that PEG-AuNP affect the myotube physiology leading these cells to be more susceptible to death stimuli in the presence of staurosporine. Altogether, these results present evidence that PEG-AuNP affect the susceptibility to apoptosis of muscle cells, contributing to development of safer strategies for intramuscular delivery.
金纳米颗粒（AuNP）已被广泛应用于许多领域，包括作为生物载体。更好地了解AuNP对肌肉细胞的安全性至关重要，因为它可能成为纳米医学领域的一种潜在工具。在此，我们描述了聚乙二醇包覆的金纳米颗粒（PEG-AuNP）与分化的骨骼肌C2C12细胞相互作用对细胞活力、线粒体功能、与存活相关的细胞信号传导、细胞因子水平以及凋亡易感性的影响。通过扫描透射电子显微镜（STEM-in-SEM）在肌管细胞中证实了直径为4.5 nm的PEG-AuNP的细胞内定位。细胞毒性分析方法表明，PEG-AuNP不影响细胞活力，但细胞内ATP水平和线粒体膜电位升高。细胞外调节蛋白激酶（ERK）的磷酸化未改变，但磷酸化蛋白激酶B（p-AKT）水平降低（p<0.01）。用PEG-AuNP预处理细胞后再用星形孢菌素诱导，会增加半胱天冬酶-3/7的活性。事实上，细胞因子分析显示，PEG-AuNP处理后，干扰素-γ（IFN-γ）和转化生长因子-β1（TGF-β1）水平急剧升高，表明炎症和纤维化表型过程被激活。这些数据表明，PEG-AuNP影响肌管生理，导致这些细胞在存在星形孢菌素的情况下更容易受到死亡刺激。总之，这些结果表明PEG-AuNP影响肌肉细胞对凋亡的易感性，有助于制定更安全的肌肉内递送策略。
