Non-toxic engineered carbon nanodiamond concentrations induce oxidative/nitrosative stress, imbalance of energy metabolism, and mitochondrial dysfunction in microglial and alveolar basal epithelial cells
Non-toxic engineered carbon nanodiamond concentrations induce oxidative/nitrosative stress, imbalance of energy metabolism, and mitochondrial dysfunction in microglial and alveolar basal epithelial cells
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DOI：
10.1038/s41419-018-0280-z
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发表时间：
2018-02-14
影响因子：
9
通讯作者：
Caruso, Giuseppe
中科院分区：
文献类型：
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作者：
Fresta, Claudia G.;Chakraborty, Aishik;Caruso, Giuseppe
工程纳米粒子正在发现广泛的生物医学应用，包括药物输送和触发细胞毒性现象的能力，可能对肿瘤细胞有用。必须充分了解它们的生物安全性和与细胞过程的相互作用。使用小胶质细胞（BV-2）和肺泡基底上皮细胞（A549），在这项研究中，我们确定了工程碳纳米金刚石（ECNs）对细胞活力，一氧化氮（NO）和活性氧（ROS）的生产，以及对能量代谢的影响。特别地，我们最初测量了细胞活力随ECN剂量增加而降低，在两种细胞系中发现了相似的细胞毒性ECN效应。随后，使用明显无细胞毒性的ECN浓度（2 μ g/mL，导致细胞数量减少< 5%），我们测定了NO和ROS的产生，并测量了与能量代谢、线粒体功能、氧化还原反应和抗氧化防御相关的化合物的浓度。我们发现，在两种细胞系中，非细胞毒性ECN浓度增加了持续氧化/亚硝化应激的NO和ROS产生，导致能量代谢失衡（高能磷酸盐和烟碱辅酶减少）和线粒体功能障碍这些结果强调了深入研究ECNs相互作用后发生的分子和生化变化的重要性（和一般的纳米颗粒）与活细胞，即使在明显无毒的浓度。由于ECNs在生物医学领域的使用越来越受到关注，因此在这些极具前景的工具找到正确的应用之前，必须对其生物安全性，毒性和/或可能的副作用进行全面评估。
Engineered nanoparticles are finding a wide spectrum of biomedical applications, including drug delivery and capacity to trigger cytotoxic phenomena, potentially useful against tumor cells. The full understanding of their biosafety and interactions with cell processes is mandatory. Using microglial (BV-2) and alveolar basal epithelial (A549) cells, in this study we determined the effects of engineered carbon nanodiamonds (ECNs) on cell viability, nitric oxide (NO) and reactive oxygen species (ROS) production, as well as on energy metabolism. Particularly, we initially measured decrease in cell viability as a function of increasing ECNs doses, finding similar cytotoxic ECN effects in the two cell lines. Subsequently, using apparently non-cytotoxic ECN concentrations (2 mu g/mL causing decrease in cell number < 5%) we determined NO and ROS production, and measured the concentrations of compounds related to energy metabolism, mitochondrial functions, oxido-reductive reactions, and antioxidant defences. We found that in both cell lines non-cytotoxic ECN concentrations increased NO and ROS production with sustained oxidative/nitrosative stress, and caused energy metabolism imbalance (decrease in high energy phosphates and nicotinic coenzymes) and mitochondrial malfunctioning (decrease in ATP/ADP ratio).These results underline the importance to deeply investigate the molecular and biochemical changes occurring upon the interaction of ECNs (and nanoparticles in general) with living cells, even at apparently non-toxic concentration. Since the use of ECNs in biomedical field is attracting increasing attention the complete evaluation of their biosafety, toxicity and/or possible side effects both in vitro and in vivo is mandatory before these highly promising tools might find the correct application.