Department of Medicine, Surgery and Neuroscience, Rheumatology Unit, University of Siena, Italy.
J Appl Toxicol. 2013 Dec;33(12):1506-13. doi: 10.1002/jat.2912. Epub 2013 Sep 11.
The aim of our study was to evaluate the effects of gold (Au) and silver (Ag) nanoparticles (NPs) at different concentrations on cultured human osteoarthritic chondrocytes. Cell viability and inducible nitric oxide synthase expression were evaluated by light microscopy. Using transmission electron microscopy (TEM) and field emission gun-based scanning transmission electron microscopy/energy dispersive spectroscopy (FEG-STEM/EDS) allowed us to localize NPs. Gene expression of matrix metalloproteinases 1, 3 and 13 and A disintegrin and metalloproteinase with thrombospondin motifs -4 and -5 were carried out by real-time polymerase chain reaction. A cell viability test indicated a significant dose-dependent cytotoxic effect of both NPs. At concentrations of 160 and 250 μM NP light microscopy showed chondrocytes with signs of apoptosis and an increased presence of inducible nitric oxide synthase. Au-NPs were characterized by FEG-STEM/EDS and TEM analysis localized NPs in cytoplasm and in endocytotic vesicles. On the contrary, the Ag-NPs were undetectable by FEG-STEM/EDS and TEM. Increased gene expression, particularly in matrix metalloproteinase-3, was observed for both NPs (160 μM), but at a concentration of 250 μM the expression of the evaluated genes became lower. Our in vitro studies, although preliminary, suggest that engineered Au and Ag-NPs appear to be harmful for human osteoarthritic chondrocytes in high concentrations (160-250 μM).
本研究旨在评估不同浓度金(Au)和银(Ag)纳米颗粒(NPs)对培养的人骨性关节炎软骨细胞的影响。通过光镜评估细胞活力和诱导型一氧化氮合酶的表达。使用透射电子显微镜(TEM)和场发射枪扫描透射电子显微镜/能量色散光谱(FEG-STEM/EDS)使我们能够定位 NPs。通过实时聚合酶链反应进行基质金属蛋白酶 1、3 和 13 以及解整合素和金属蛋白酶与血栓反应蛋白 4 和 5 的基因表达。细胞活力测试表明,两种 NPs 均具有显著的剂量依赖性细胞毒性作用。在 160 和 250 μM NP 浓度下,光镜显示软骨细胞具有凋亡迹象,诱导型一氧化氮合酶的表达增加。FEG-STEM/EDS 和 TEM 分析表明 Au-NPs 定位于细胞质和内吞小泡中。相比之下,Ag-NPs 无法通过 FEG-STEM/EDS 和 TEM 检测到。两种 NPs(160 μM)都观察到基因表达增加,特别是基质金属蛋白酶-3,但在 250 μM 浓度下,评估基因的表达降低。我们的体外研究虽然初步,但表明工程 Au 和 Ag-NPs 在高浓度(160-250 μM)下似乎对人骨性关节炎软骨细胞有害。