TY - JOUR A1 - Haase, A. A1 - Rott, S. A1 - Mantion, Alexandre A1 - Graf, P. A1 - Plendl, J. A1 - Thünemann, Andreas A1 - Meier, W.P. A1 - Taubert, A. A1 - Luch, A. A1 - Reiser, G T1 - Effects of silver nanoparticles on primary mixed neural cell cultures: uptake, oxidative stress and acute calcium responses JF - Toxicological sciences N2 - In the body, nanoparticles can be systemically distributed and then may affect secondary target organs, such as the central nervous system (CNS). Putative adverse effects on the CNS are rarely investigated to date. Here, we used a mixed primary cell model consisting mainly of neurons and astrocytes and a minor proportion of oligodendrocytes to analyze the effects of well-characterized 20 and 40 nm silver nanoparticles (SNP). Similar gold nanoparticles served as control and proved inert for all endpoints tested. SNP induced a strong size-dependent cytotoxicity. Additionally, in the low concentration range (up to 10 µg/ml of SNP), the further differentiated cultures were more sensitive to SNP treatment. For detailed studies, we used low/medium dose concentrations (up to 20 µg/ml) and found strong oxidative stress responses. Reactive oxygen species (ROS) were detected along with the formation of protein carbonyls and the induction of heme oxygenase-1. We observed an acute calcium response, which clearly preceded oxidative stress responses. ROS formation was reduced by antioxidants, whereas the calcium response could not be alleviated by antioxidants. Finally, we looked into the responses of neurons and astrocytes separately. Astrocytes were much more vulnerable to SNP treatment compared with neurons. Consistently, SNP were mainly taken up by astrocytes and not by neurons. Immunofluorescence studies of mixed cell cultures indicated stronger effects on astrocyte morphology. Altogether, we can demonstrate strong effects of SNP associated with calcium dysregulation and ROS formation in primary neural cells, which were detectable already at moderate dosages. KW - Silver nanoparticles KW - Neurons KW - Oxidative stress KW - Protein carbonyls KW - Calcium KW - Reference material KW - Nanoparticle KW - Small-angle X-ray scattering KW - SAXS PY - 2012 DO - https://doi.org/10.1093/toxsci/kfs003 SN - 1096-6080 SN - 1096-0929 VL - 126 IS - 2 SP - 457 EP - 468 PB - Oxford University Press CY - Oxford AN - OPUS4-25633 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Haase, A. A1 - Tentschert, J. A1 - Jungnickel, H. A1 - Graf, P. A1 - Mantion, Alexandre A1 - Draude, F. A1 - Plendl, J. A1 - Goetz, M.E. A1 - Galla, S. A1 - Masic, A. A1 - Thünemann, Andreas A1 - Taubert, A. A1 - Arlinghaus, H. F. A1 - Luch, A. T1 - Toxicity of silver nanoparticles in human macrophages: uptake, intracellular distribution and cellular responses JF - Journal of physics / Conference series N2 - Silver nanoparticles (SNP) are among the most commercialized nanoparticles worldwide. They can be found in many diverse products, mostly because of their antibacterial properties. Despite its widespread use only little data on possible adverse health effects exist. It is difficult to compare biological data from different studies due to the great variety in sizes, coatings or shapes of the particles. Here, we applied a novel synthesis approach to obtain SNP, which are covalently stabilized by a small peptide. This enables a tight control of both size and shape. We applied these SNP in two different sizes of 20 or 40 nm (Ag20Pep and Ag40Pep) and analyzed responses of THP-1-derived human macrophages. Similar gold nanoparticles with the same coating (Au20Pep) were used for comparison and found to be non-toxic. We assessed the cytotoxicity of particles and confirmed their cellular uptake via transmission electron microscopy and confocal Raman microscopy. Importantly a majority of the SNP could be detected as individual particles spread throughout the cells. Furthermore we studied several types of oxidative stress related responses such as induction of heme oxygenase I or formation of protein carbonyls. In summary, our data demonstrate that even low doses of SNP exerted adverse effects in human macrophages. KW - Silver nanoparticles KW - Neurotoxicology KW - Protein carbonyls KW - ROS PY - 2011 DO - https://doi.org/10.1088/1742-6596/304/1/012030 SN - 1742-6588 SN - 1742-6596 VL - 304 SP - 012030-1 - 012030-14 PB - IOP Publ. CY - Bristol, UK AN - OPUS4-24035 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Haase, A. A1 - Arlinghaus, H. F. A1 - Tentschert, J. A1 - Jungnickel, H. A1 - Graf, P. A1 - Mantion, Alexandre A1 - Draude, F. A1 - Galla, S. A1 - Plendl, J. A1 - Goetz, M.E. A1 - Masic, A. A1 - Meier, W. A1 - Thünemann, Andreas A1 - Taubert, A. A1 - Luch, A. T1 - Application of laser postionization secondary neutral mass spectrometry / time-of-flight secondary ion mass spectrometry in nanotoxicology: Visualization of nanosilver in human macrophages and cellular responses JF - ACS nano N2 - Silver nanoparticles (SNP) are the subject of worldwide commercialization because of their antimicrobial effects. Yet only little data on their mode of action exist. Further, only few techniques allow for visualization and quantification of unlabeled nanoparticles inside cells. To study SNP of different sizes and coatings within human macrophages, we introduce a novel laser postionization secondary neutral mass spectrometry (Laser-SNMS) approach and prove this method superior to the widely applied confocal Raman and transmission electron microscopy. With time-of-flight secondary ion mass spectrometry (TOF-SIMS) we further demonstrate characteristic fingerprints in the lipid pattern of the cellular membrane indicative of oxidative stress and membrane fluidity changes. Increases of protein carbonyl and heme oxygenase-1 levels in treated cells confirm the presence of oxidative stress biochemically. Intriguingly, affected phagocytosis reveals as highly sensitive end point of SNP-mediated adversity in macrophages. The cellular responses monitored are hierarchically linked, but follow individual kinetics and are partially reversible. KW - Nanosilver KW - Laser-SNMS KW - TOF-SIMS KW - Confocal Raman microscopy KW - Oxidative stress KW - Protein carbonyls PY - 2011 DO - https://doi.org/10.1021/nn200163w SN - 1936-0851 VL - 5 IS - 4 SP - 3059 EP - 3068 PB - ACS Publ. CY - Washington, DC, USA AN - OPUS4-23656 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -