Department of Animal, Dairy and Veterinary Sciences, Utah State University, Logan, UT, 84322, USA; Institute of Animal Husbandry and Veterinary Science, Guizhou Academy of Agricultural Sciences, Guiyang, Guizhou, 550005, China.
Department of Biological Engineering, Utah State University, Logan, UT, 84322, USA.
Talanta. 2022 Aug 15;246:123522. doi: 10.1016/j.talanta.2022.123522. Epub 2022 May 10.
Since silver ion is known for its antimicrobial function, most of the research has focused mainly on toxicity effects rather than the role of silver ion in general biology and the behind mechanism of actions of silver ion in mammalian cells. Moreover, a conventional in vitro approach to estimate the effects of silver ion on cells does not provide information about the biochemical changes and might accompany artifacts due to invasive and destructive sample preparation processes. In the present study, in-situ real time approaches were applied to evaluate the impact of silver ion (0.57, 1.34, 1.96, 2.33 mg/L) on fibroblast cells. Raman spectroscopy analysis showed that Raman peak intensities of proteins and nucleic acids significantly increased in the cells after exposure to silver ion for 21 h, especially at relatively higher levels 1.34, 1.96, and 2.33 mg/L. Raman peak at 1585 cm and liquid scanning transmission electron microscopy energy-dispersive x-ray spectroscopy (STEM-EDS) analysis revealed the fate of silver ion that was taken up by the cell and reduced into metallic silver accumulating in the cell as silver nanoparticles. These results suggest cells were undergoing different activities such as enhanced metabolic activities rather than cell apoptosis or cell death. Additionally, Raman spectroscopy predicted the level of silver ion exposed to the cell at 2.11 ± 0.38 and 1.73 ± 0.26 mg/L by the PLS prediction model, compared with the results measured by inductively coupled plasma mass spectrometry (ICP-MS), 2.14 ± 0.07 and 1.87 ± 0.07 mg/L respectively, suggesting Raman spectroscopy can provide a new and fast approach to determine and measure the concentration of silver ion or probably other tested molecules treated to the cell for the future research.
由于银离子具有抗菌功能，因此大多数研究主要集中在毒性作用上，而不是银离子在一般生物学中的作用及其在哺乳动物细胞中的作用机制。此外，传统的体外方法来估计银离子对细胞的影响不能提供关于生化变化的信息，并且由于侵入性和破坏性的样品制备过程，可能会伴随出现假象。在本研究中，应用了原位实时方法来评估银离子（0.57、1.34、1.96、2.33mg/L）对成纤维细胞的影响。拉曼光谱分析表明，暴露于银离子 21 小时后，细胞中蛋白质和核酸的拉曼峰强度显着增加，特别是在相对较高的 1.34、1.96 和 2.33mg/L 水平下。在 1585cm 处的拉曼峰和液体扫描透射电子显微镜能量色散 X 射线能谱（STEM-EDS）分析揭示了银离子的命运，细胞摄取的银离子被还原成纳米银并在细胞内积累。这些结果表明，细胞正在经历不同的活动，例如增强的代谢活性，而不是细胞凋亡或细胞死亡。此外，通过偏最小二乘预测模型，拉曼光谱预测了细胞暴露于 2.11±0.38 和 1.73±0.26mg/L 的银离子水平，而电感耦合等离子体质谱（ICP-MS）测量的结果分别为 2.14±0.07 和 1.87±0.07mg/L，表明拉曼光谱可以为未来的研究提供一种新的快速方法来确定和测量细胞中银离子或可能其他测试分子的浓度。
