Proinflammatory and cytotoxic response to nanoparticles in precision-cut lung slices

• Stephanie Hirn,
• Nadine Haberl,
• Kateryna Loza,
• Matthias Epple,
• Wolfgang G. Kreyling,
• Barbara Rothen-Rutishauser,
• Markus Rehberg and
• Fritz Krombach

Beilstein J. Nanotechnol. 2014, 5, 2440–2449, doi:10.3762/bjnano.5.253

• microscopy was employed to assess the localization of Ag-NPs in PCLS after 24 h of incubation. Exposure of PCLS to ZnO-NPs for 4 and 24 h resulted in a strong decrease in cell viability, while quartz particles had no cytotoxic effect. Moreover, only a slight cytotoxic response was detected by LDH release

• investigate whether PCLS have the potential to serve as a generally applicable effective tool in nanotoxicology. First, we determined the viability of PCLS up to 72 h by carrying out live/dead staining, lactate dehydrogenase (LDH) assay, and WST-1 assay. Second, we assessed the cytotoxic and proinflammatory

• response of PCLS to Ag-NPs through LDH assay, WST-1 assay, and ELISA assay for CXCL-1 and tumour necrosis factor-α (TNF-α) release. ZnO-NPs served as a soluble control and quartz particles as a non-soluble control, since both particle types have induced cytotoxic as well as inflammatory responses in in

Effects of surface functionalization on the adsorption of human serum albumin onto nanoparticles – a fluorescence correlation spectroscopy study

• Pauline Maffre,
• Stefan Brandholt,
• Karin Nienhaus,
• Li Shang,
• Wolfgang J. Parak and
• G. Ulrich Nienhaus

Beilstein J. Nanotechnol. 2014, 5, 2036–2047, doi:10.3762/bjnano.5.212

• taken on a time-resolved microscopy system (Microtime 200, Picoquant, Berlin, Germany) using an inverted epi-fluorescence microscope (IX71, Olympus). Here, the sample is excited by a pulsed, solid state, 530 nm laser (LDH-P-FA-530, Picoquant), sent into the water-immersion objective (UPLSAPO 60XW

PVP-coated, negatively charged silver nanoparticles: A multi-center study of their physicochemical characteristics, cell culture and in vivo experiments

• Sebastian Ahlberg,
• Alexandra Antonopulos,
• Jörg Diendorf,
• Ralf Dringen,
• Matthias Epple,
• Rebekka Flöck,
• Wolfgang Goedecke,
• Christina Graf,
• Nadine Haberl,
• Jens Helmlinger,
• Fabian Herzog,
• Frederike Heuer,
• Stephanie Hirn,
• Christian Johannes,
• Stefanie Kittler,
• Manfred Köller,
• Katrin Korn,
• Wolfgang G. Kreyling,
• Fritz Krombach,
• Jürgen Lademann,
• Kateryna Loza,
• Eva M. Luther,
• Marcelina Malissek,
• Martina C. Meinke,
• Daniel Nordmeyer,
• Anne Pailliart,
• Jörg Raabe,
• Fiorenza Rancan,
• Barbara Rothen-Rutishauser,
• Eckart Rühl,
• Carsten Schleh,
• Andreas Seibel,
• Christina Sengstock,
• Lennart Treuel,
• Annika Vogt,
• Katrin Weber and
• Reinhard Zellner

Beilstein J. Nanotechnol. 2014, 5, 1944–1965, doi:10.3762/bjnano.5.205

• PCLS to silver nanoparticles (10, 20 and 30 µg mL−1) under submerged conditions for 4 and 24 h resulted in only weak cytotoxicity (LDH release), but did not induce a proinflammatory response (CXCL-1 and TNF-α release). Interestingly, multiphoton microscopy revealed that the silver nanoparticles were

In vitro and in vivo interactions of selected nanoparticles with rodent serum proteins and their consequences in biokinetics

• Wolfgang G. Kreyling,
• Stefanie Fertsch-Gapp,
• Martin Schäffler,
• Blair D. Johnston,
• Nadine Haberl,
• Christian Pfeiffer,
• Jörg Diendorf,
• Carsten Schleh,
• Stephanie Hirn,
• Manuela Semmler-Behnke,
• Matthias Epple and
• Wolfgang J. Parak

Beilstein J. Nanotechnol. 2014, 5, 1699–1711, doi:10.3762/bjnano.5.180

• twenty-four hours the viability of the PCLS was tested by lactate dehydrogenase (LDH) analysis for cell membrane damage and a WST-1 assay for mitochondrial activity in the cell culture supernatant as well as the release of the pro-inflammatory cytokines tumor necrosis factor alpha (TNF-α) and interleukin

• -inflammatory cytokines of PCLS exposed to either AgNP or dissolved Ag or mixtures of both. The LDH release of PCLS increased significantly when the rats were instilled with the aqueous supernatants of AgNP centrifugation compared to PCLS of untreated control rats (Figure 7). Consistently, LDH was also

• /digested intracellularly probably faster than the engulfed AgNP that formed Ag+ ions or other reactive Ag species, which apparently caused the increased LDH release and the release of both cytokines [9][27]. Interestingly, instilled silver acetate showed a similar trend as the suspension of the six-month

Different endocytotic uptake mechanisms for nanoparticles in epithelial cells and macrophages

• Dagmar A. Kuhn,
• Dimitri Vanhecke,
• Benjamin Michen,
• Fabian Blank,
• Peter Gehr,
• Alke Petri-Fink and
• Barbara Rothen-Rutishauser

Beilstein J. Nanotechnol. 2014, 5, 1625–1636, doi:10.3762/bjnano.5.174

• dehydrogenase (LDH) assay (see Figure S1, Supporting Information File 1). Trypan blue staining marked the integrity of the cell membrane for cells which were impaired by the inhibitor (red insets, Figure 3) and revealed the following percentage of dead cells for J774A.1 cells (n = 3): negative control 20% (SD

• study the uptake of the two different particle types. Visualization of the fluorescently tagged particles was done by LSM. LDH measurements revealed no cytotoxicity for the combined inhibitors and endocytotic protein markers for both cell types being analyzed. Additionally, inhibitors which negatively

• . Image acquisition was performed as described above. Lactate dehydrogenase (LDH) assay A 1 mL sample of the supernatant of each experiment was collected and stored at 4 °C to determine cytotoxicity. Triton X (0.2% in unsupplemented RPMI) was used for cell lysis as a positive control. The supernatant of

Silica nanoparticles are less toxic to human lung cells when deposited at the air–liquid interface compared to conventional submerged exposure

• Alicja Panas,
• Andreas Comouth,
• Harald Saathoff,
• Thomas Leisner,
• Marco Al-Rawi,
• Michael Simon,
• Gunnar Seemann,
• Olaf Dössel,
• Sonja Mülhopt,
• Hanns-Rudolf Paur,
• Susanne Fritsch-Decker,
• Carsten Weiss and
• Silvia Diabaté

Beilstein J. Nanotechnol. 2014, 5, 1590–1602, doi:10.3762/bjnano.5.171

• Information File 1). Figure 7 shows that the electrical field and the exposure to filtered air had no effect on the release of lactate dehydrogenase (LDH) into the medium. Enhanced levels of LDH indicate membrane damage, which leads to cell death. LDH values of the ALI-exposed control samples were similar to

• those detected under submerged conditions. In contrast, exposure to silica NPs induced strong LDH release under submerged exposure conditions. The LDH release after submerged exposure to 15.6 µg·cm−2 (50 µg·mL−1) Aerosil200 indicates 100% cell lysis as it was comparable to the LDH release from Triton X

• . Exposure to silica NPs induced a strong release of IL-8 under submerged exposure conditions. Cells exposed to NPs at the ALI, however, released much lower IL-8 levels. Again under both conditions, as also seen for the release of LDH above, SiO2-50 nm NPs were less effective than Aerosil200 NPs. IL-6

Mimicking exposures to acute and lifetime concentrations of inhaled silver nanoparticles by two different in vitro approaches

• Fabian Herzog,
• Kateryna Loza,
• Sandor Balog,
• Martin J. D. Clift,
• Matthias Epple,
• Peter Gehr,
• Alke Petri-Fink and
• Barbara Rothen-Rutishauser

Beilstein J. Nanotechnol. 2014, 5, 1357–1370, doi:10.3762/bjnano.5.149

• (diameter 100 nm; coated with polyvinylpyrrolidone: PVP). Ag NPs were found to be highly aggregated within ALI exposed cells with no impairment of cell morphology. Furthermore, a significant increase in release of cytotoxic (LDH), oxidative stress (SOD-1, HMOX-1) or pro-inflammatory markers (TNF-α, IL-8

• measured the release of lactate dehydrogenase (LDH) as a marker for the cell integrity to assess the cytotoxic potential of cells exposed to Ag NPs. Also included were cultures incubated with lipopolysaccharide (LPS) as a pro-inflammatory stimulus and tumor necrosis factor alpha (TNF-α) as positive control

• Supporting Information File 1 (Figure S1 and Figure S2). Cells exposed at the ALI did not show a significant LDH release for all three deposited Ag NP concentrations of 0.03, 0.3, and 3 µg Ag/cm2 4 and 24 h after exposure (Figure 4A). However, elevated levels of LDH were observed at 3 µg Ag/cm2 after 24 h

Cytotoxic and proinflammatory effects of PVP-coated silver nanoparticles after intratracheal instillation in rats

• Nadine Haberl,
• Stephanie Hirn,
• Alexander Wenk,
• Jörg Diendorf,
• Matthias Epple,
• Blair D. Johnston,
• Fritz Krombach,
• Wolfgang G. Kreyling and
• Carsten Schleh

Beilstein J. Nanotechnol. 2013, 4, 933–940, doi:10.3762/bjnano.4.105

• rats. Monodisperse, PVP-coated AgNP (70 nm) showing little agglomeration in aqueous suspension were instilled intratracheally. After 24 hours, the lungs were lavaged, and lactate dehydrogenase (LDH), total protein, and cytokine levels as well as total and differential cell counts were measured in the

• bronchoalveolar lavage fluid (BALF). Instillation of 50 µg PVP-AgNP did not result in elevated LDH, total protein, or cytokine levels in BALF compared to the control, whereas instillation of 250 µg PVP-AgNP caused a significant increase in LDH (1.9-fold) and total protein (1.3-fold) levels as well as in

• intratracheally into healthy rats, and cytotoxic and proinflammatory effects were determined by measuring lactate dehydrogenase (LDH), protein, and cytokine levels as well as total and differential cell counts in bronchoalveolar lavage fluid (BALF). Results Particle characterization The mean diameter of the PVP

Selective surface modification of lithographic silicon oxide nanostructures by organofunctional silanes

• Thomas Baumgärtel,
• Christian von Borczyskowski and
• Harald Graaf

Beilstein J. Nanotechnol. 2013, 4, 218–226, doi:10.3762/bjnano.4.22

• subsequent cleaning in ultrasonic baths of acetone, dichloromethane and ethanol. All silanization experiments were carried out at room temperature (21 °C). Fluorescence investigations of the samples were performed with a home-built microscope setup. The 465 nm excitation light from a pulsed laser diode (”LDH

Characterization of protein adsorption onto FePt nanoparticles using dual-focus fluorescence correlation spectroscopy

• Pauline Maffre,
• Karin Nienhaus,
• Faheem Amin,
• Wolfgang J. Parak and
• G. Ulrich Nienhaus

Beilstein J. Nanotechnol. 2011, 2, 374–383, doi:10.3762/bjnano.2.43

• . 2fFCS setup The 2fFCS setup is based on a time-resolved confocal microscopy system (Microtime 200, PicoQuant, Berlin, Germany). Instead of using a single excitation laser, the light from two identical, orthogonally polarized pulsed 640 nm diode lasers (LDH-P-C-640B, Picoquant, Berlin, Germany) was