Milić Mirta, Leitinger Gerd, Pavičić Ivan, Zebić Avdičević Maja, Dobrović Slaven, Goessler Walter, Vinković Vrček Ivana
Institute for Medical Research and Occupational Health, Ksaverska cesta 2, 10 000, Zagreb, Croatia.
J Appl Toxicol. 2015 Jun;35(6):581-92. doi: 10.1002/jat.3081. Epub 2014 Oct 28.
The rapid progress and early commercial acceptance of silver-based nanomaterials is owed to their biocidal activity. Besides embracing the antimicrobial potential of silver nanoparticles (AgNPs), it is imperative to give special attention to the potential adverse health effects of nanoparticles owing to prolonged exposure. Here, we report a detailed study on the in vitro interactions of citrate-coated AgNPs with porcine kidney (Pk15) cells. As uncertainty remains whether biological/cellular responses to AgNPs are solely as a result of the release of silver ions or whether the AgNPs themselves have toxic effects, we investigated the effects of Ag(+) on Pk15 cells for comparison. Next, we investigated the cellular uptake of both AgNPs and Ag(+) in Pk15 cells at various concentrations applied. The detected Ag contents in cells exposed to 50 mg l(-1) AgNPs and 50 mg l(-1) Ag(+) were 209 and 25 µg of Ag per 10(6) cells, respectively. Transmission electron microscopy (TEM) images indicated that the Pk15 cells internalized AgNPs by endocytosis. Both forms of silver, nano and ionic, decreased the number of viable Pk15 cells after 24 h in a dose-dependent manner. In spite of a significant uptake into the cells, AgNPs had only insignificant toxicity at concentrations lower than 25 mg l(-1) , whereas Ag(+) exhibited a significant decrease in cell viability at one-fifth of this concentration. The Comet assay suggested that a rather high concentration of AgNP (above 25 mg l(-1) ) is able to induce genotoxicity in Pk15 cells. Further studies must seek deeper understanding of AgNP behavior in biological media and their interactions with cellular membranes.
银基纳米材料的迅速发展及其早期在商业上的被接受归功于它们的杀菌活性。除了利用银纳米颗粒(AgNPs)的抗菌潜力外,由于长期接触,必须特别关注纳米颗粒对健康的潜在不利影响。在此,我们报告了一项关于柠檬酸盐包覆的AgNPs与猪肾(Pk15)细胞体外相互作用的详细研究。由于对于AgNPs的生物/细胞反应是仅仅由于银离子的释放,还是AgNPs本身具有毒性作用仍存在不确定性,我们研究了Ag(+)对Pk15细胞的影响以作比较。接下来,我们研究了在不同应用浓度下Pk15细胞对AgNPs和Ag(+)的细胞摄取情况。暴露于50 mg l(-1) AgNPs和50 mg l(-1) Ag(+)的细胞中检测到的Ag含量分别为每10(6)个细胞209和25 μg Ag。透射电子显微镜(TEM)图像表明Pk15细胞通过内吞作用内化了AgNPs。纳米银和离子银这两种形式在24小时后均以剂量依赖的方式减少了存活的Pk15细胞数量。尽管细胞对AgNPs有大量摄取,但在浓度低于25 mg l(-1)时AgNPs仅具有微不足道的毒性,而Ag(+)在该浓度的五分之一时就表现出细胞活力的显著下降。彗星试验表明,相当高浓度的AgNP(高于25 mg l(-1))能够在Pk15细胞中诱导遗传毒性。进一步的研究必须更深入地了解AgNP在生物介质中的行为及其与细胞膜的相互作用。