Oxygen-plasma-modified biomimetic nanofibrous scaffolds for enhanced compatibility of cardiovascular implants

• Anna Maria Pappa,
• Varvara Karagkiozaki,
• Silke Krol,
• Spyros Kassavetis,
• Dimitris Konstantinou,
• Charalampos Pitsalidis,
• Lazaros Tzounis,
• Nikos Pliatsikas and
• Stergios Logothetidis

Beilstein J. Nanotechnol. 2015, 6, 254–262, doi:10.3762/bjnano.6.24

• to the reference wavelength of 690 nm. Data (n = 3) were presented as means of O.D. values as well as normalized according to the control and presented as % cell viability. Optical imaging through methylene blue staining: Once fibroblasts were seeded onto either unmodified or surface-modified

Mechanical properties of MDCK II cells exposed to gold nanorods

• Anna Pietuch,
• Bastian Rouven Brückner,
• David Schneider,
• Marco Tarantola,
• Christina Rosman,
• Carsten Sönnichsen and
• Andreas Janshoff

Beilstein J. Nanotechnol. 2015, 6, 223–231, doi:10.3762/bjnano.6.21

• number of CTAB molecules which can, in principle, be released from the particle surface assuming a concentration of 12 μg/mL, we only found a Young’s modulus of E = 6.5 ± 1 kPa. Fluorescence microscopy images (staining of actin and microtubules) as well as AFM images (topography) reveal that the cells

• with PBS. Incubation with staining solution was carried out following the manufacture’s recommendation. For F-actin staining Alexa Fluor 546 Phalloidin (Invitrogen, Darmstadt, Germany) and for microtubules labeling Alexa 488 conjugated mouse anti-β-tubulin (BD Biosciences, Heidelberg, Germany) was used

• . Nucleus staining was carried out with DAPI (4’,6-diamidino-2-phenylindol, 50 ng/mL in PBS) (Sigma-Aldrich, Steinheim, Germany). Dark-field microscopy was carried out with an upright microscope with dark-field condensor (Olympus BX51) equipped with a 40× water immersion objective. AFM imaging and force

Multifunctional layered magnetic composites

• Maria Siglreitmeier,
• Baohu Wu,
• Tina Kollmann,
• Martin Neubauer,
• Gergely Nagy,
• Dietmar Schwahn,
• Vitaliy Pipich,
• Damien Faivre,
• Dirk Zahn,
• Andreas Fery and
• Helmut Cölfen

Beilstein J. Nanotechnol. 2015, 6, 134–148, doi:10.3762/bjnano.6.13

• to the matrix, which might derive from the usage of staining media or dehydration. For comparison studies, the structure of the original nacre matrix (Haliotis laevigata) was analyzed as well. Figure 2 represents very-small (VSANS) and small (SANS) angle neutron scattering profiles of nacre (top) and

• studies no staining of proteins in between the layers could be observed. Therefore we conclude that the insoluble matrix proteins are dominantly located directly at the β-chitin matrix and are not present in between the layers. Figure 3c and Figure 3d show an embedded sample of insoluble nacre matrix

• infiltrated with gelatin by a vacuum infiltration process. Staining of this sample illustrates not only blue stained chitin layers and insoluble matrix proteins but also colored areas in between the layers, indicating a filling of the matrix with gelatin. The interaction and positive stain of gelatin and

Mammalian cell growth on gold nanoparticle-decorated substrates is influenced by the nanoparticle coating

• Christina Rosman,
• Sebastien Pierrat,
• Marco Tarantola,
• David Schneider,
• Eva Sunnick,
• Andreas Janshoff and
• Carsten Sönnichsen

Beilstein J. Nanotechnol. 2014, 5, 2479–2488, doi:10.3762/bjnano.5.257

• staining test for live/dead should be performed. This was not performed since our results already indicated that CTAB nanorods are not suitable for live cell applications due to their impact on the native cell behavior. However, although the proliferation behavior of the tracked cells was poor, active cell

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

• , and dendritic cells, in their natural context has been shown in several studies by immunohistochemical staining [7][8][9]. Nevertheless, this in vitro system has also its limitations. There are obvious disadvantages in comparison to the in vivo situation such as no ventilation, no stretching, and no

• 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

• staining, and WST-1 assay. As LDH is present in the cytoplasm of cells, detection of LDH in the culture medium of PCLS indicates a loss of cell membrane integrity. Therefore, LDH release is a direct measure of a cytotoxic response or an indirect measure of cell viability. As displayed in Figure 1, the LDH

Nanoparticle interactions with live cells: Quantitative fluorescence microscopy of nanoparticle size effects

• Li Shang,
• Karin Nienhaus,
• Xiue Jiang,
• Linxiao Yang,
• Katharina Landfester,
• Volker Mailänder,
• Thomas Simmet and
• G. Ulrich Nienhaus

Beilstein J. Nanotechnol. 2014, 5, 2388–2397, doi:10.3762/bjnano.5.248

• NPs in each cell was obtained by normalizing the integrated intensity by the cell area. The cell membrane and the intracellular region were identified based on the membrane staining. Schematic representation of the cellular uptake of (a) large and (b) small NPs. Whereas larger NPs exert interactions

Coating with luminal gut-constituents alters adherence of nanoparticles to intestinal epithelial cells

• Heike Sinnecker,
• Katrin Ramaker and
• Andreas Frey

Beilstein J. Nanotechnol. 2014, 5, 2308–2315, doi:10.3762/bjnano.5.239

• grown on transwell filters (Figure 3A, B; counter-staining of the cell surface with a lectin). Pretreatment with the protein BSA appears to alleviate agglomeration as well as adherence. Only few distinct spots are visible on the cell surface, compared to a pronounced adherence of markedly larger

Effect of silver nanoparticles on human mesenchymal stem cell differentiation

• Christina Sengstock,
• Jörg Diendorf,
• Matthias Epple,
• Thomas A. Schildhauer and
• Manfred Köller

Beilstein J. Nanotechnol. 2014, 5, 2058–2069, doi:10.3762/bjnano.5.214

• used (Figure 2B). To confirm adipogenic differentiation, the lipid content of the cells was visualized by using oil red O or Bodipy493/503 staining. As shown in Figure 3C, hMSCs differentiated into adipocytes in the presence of adipogenic-differentiation media (positive control; Figure 3C), in contrast

• ) revealed a decrease in lipid vacuoles with increasing silver concentrations. This decrease was significant at the applied concentrations of 10 µg·mL−1 for Ag-NP (black bars) or 1.0 µg·mL−1 for Ag+ ions (grey bars). The differences between oil red extraction and phase analysis after Bodipy493/503 staining

• of Ag-NP/Ag+ ions on the osteogenic differentiation of hMSCs was investigated after a period of 21 d. To confirm the differentiation of the Ag-NP/Ag+ ion-treated hMSCs into osteoblasts, alizarin red S staining was carried out to verify the mineralization of the cells. hMSCs exposed to 10 µg·mL−1 Ag

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

• demonstrated for hMSC, primary T-cells, primary monocytes, and astrocytes. A visualization of particles inside cells is possible by X-ray microscopy, fluorescence microscopy, and combined FIB/SEM analysis. By staining organelles, their localization inside the cell can be additionally determined. While primary

• (TEM), this technique requires thin samples and, hence, slicing of the samples as well as additional staining, which both might change the properties of the samples. Furthermore, energy dispersive X-ray spectroscopy (EDX) combined with TEM has only limited spectral and spatial resolution and

• used as a tool to detect DNA-DSB by specific antibodies recognizing γ-H2AX. Staining of cells with antibodies directed against γ-H2AX results in a speckled staining of the nucleus. It is generally accepted that a single focus is representing a single DSB [118]. In the following, we describe experiments

Biocompatibility of cerium dioxide and silicon dioxide nanoparticles with endothelial cells

• Claudia Strobel,
• Martin Förster and
• Ingrid Hilger

Beilstein J. Nanotechnol. 2014, 5, 1795–1807, doi:10.3762/bjnano.5.190

• markers, respectively. After staining both CD31 (monoclonal anti-human CD31 antibodies conjugated to fluorescein isothiocyanate (FITC), Miltenyi Biotec GmbH, Germany) and CD90 (monoclonal anti-human CD90 antibodies conjugated to R-phycoerythrin (PE) Miltenyi Biotec GmbH, Germany), the cells were washed

• . As a permeabilization reagent, 0.1% (v/v) Saponin (Sigma-Aldrich Chemie GmbH, Germany) in Hank’s BSS was used. Intracellular staining of vWF with allophycocyanin (APC) conjugated mouse monoclonal anti-human vWF-A2 antibodies (R&D Systems, Inc., USA) followed. Unstained cells, cells stained with mouse

The surface properties of nanoparticles determine the agglomeration state and the size of the particles under physiological conditions

• Christoph Bantz,
• Olga Koshkina,
• Thomas Lang,
• Hans-Joachim Galla,
• C. James Kirkpatrick,
• Roland H. Stauber and
• Michael Maskos

Beilstein J. Nanotechnol. 2014, 5, 1774–1786, doi:10.3762/bjnano.5.188

• staining procedures might cause additional artifacts and furthermore also lead to an increase in preparation time, no staining was applied here and for the POS particles the TEM investigation is only useful at the first stage of the modification procedure. To compensate for the mentioned drawbacks of the

Influence of surface-modified maghemite nanoparticles on in vitro survival of human stem cells

• Michal Babič,
• Daniel Horák,
• Lyubov L. Lukash,
• Tetiana A. Ruban,
• Yurii N. Kolomiets,
• Svitlana P. Shpylova and
• Oksana A. Grypych

Beilstein J. Nanotechnol. 2014, 5, 1732–1737, doi:10.3762/bjnano.5.183

• medium. Confocal micrographs of 4BL human stem cells treated with (a, b) D-mannose-coated γ-Fe2O3, (c, d) PDMAAm-coated γ-Fe2O3 and (e, f) non-coated γ-Fe2O3 nanoparticles. Staining with DAPI and ThR. Scale bars: 10 μm. Acknowledgements Financial support of Ministry of Education, Youth and Sports of the

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

• protein densitometry. Only predominantly binding serum proteins could be analyzed by this quantitative approach. A typical electrophoresis gel visualized by Coomassie staining is given in Figure 2, which shows a variety of proteins present in the corona of the three differently sized AuNP. Proteins cover

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

• affected the cells are summarized in Table 1 and the images in Figure 3 are presented with either red (impairment by inhibitor) or green (no effect by inhibitor) letter insets. Trypan blue staining demonstrated the same outcome. The percentage of dead cells which were treated with cytochalasin D was not as

• , flotillin-1 and caveolin-1 (all fluorescently labelled with Alexa Fluor 488, antibodies-online GmbH, Aachen, Germany) were used at a final dilution of 1:20 in 1× PBS on fixed cells. After 1 hour of staining (in a dark room at room temperature) and three washing cycles with 1× PBS, the cells were mounted

Protein-coated pH-responsive gold nanoparticles: Microwave-assisted synthesis and surface charge-dependent anticancer activity

• Dickson Joseph,
• Nisha Tyagi,
• Christian Geckeler and
• Kurt E.Geckeler

Beilstein J. Nanotechnol. 2014, 5, 1452–1462, doi:10.3762/bjnano.5.158

• ). The BSA-AuNP sample was mixed with 2× sample buffer (Sigma) and heated at 80 °C for 15 min. Further protein was separated through a 4% stacking gel and 12% separation gel. To determine the amount of protein in the sample, the gels were stained with Coomassie blue (Sigma) staining solution for 2 h. The

Near-field photochemical and radiation-induced chemical fabrication of nanopatterns of a self-assembled silane monolayer

• Ulrich C. Fischer,
• Carsten Hentschel,
• Florian Fontein,
• Linda Stegemann,
• Christiane Hoeppener,
• Harald Fuchs and
• Stefanie Hoeppener

Beilstein J. Nanotechnol. 2014, 5, 1441–1449, doi:10.3762/bjnano.5.156

• , respectively. The 1.2 µm structure is clearly resolved in the fluorescence image, but due to the diffraction limit the 0.22 µm pattern cannot be resolved. Only the defect structures of the periodic nanopatterns are recognized. In addition to the staining of the chemical nanostructures with fluorescein

• (a) 1.2 µm latex beads and of (b) 220 nm latex beads after staining of the chemically functionalized areas with fluorescein molecules. Optically only the larger hexagonal nanopattern can be resolved, whereas the fluorescence image of the 220 nm pattern is characterized by defect structures. (c) AFM

The protein corona protects against size- and dose-dependent toxicity of amorphous silica nanoparticles

• Dominic Docter,
• Christoph Bantz,
• Dana Westmeier,
• Hajo J. Galla,
• Qiangbin Wang,
• James C. Kirkpatrick,
• Peter Nielsen,
• Michael Maskos and
• Roland H. Stauber

Beilstein J. Nanotechnol. 2014, 5, 1380–1392, doi:10.3762/bjnano.5.151

• bromophenol blue) to the pellet and incubation at 95 °C for 5 min. 1D SDS-PAGE Discontinuous SDS-polyacrylamide gel electrophoresis (PAGE) was carried out according to standard procedures [51]. Proteins were visualized by staining with Coomassie brilliant blue R-250 as described [52]. All experiments were

• cells). Nuclei were stained by addition of Hoechst 33342 at a final concentration of 40 µM for 10 min. Images were acquired and analyzed on the Cellomics ArrayScan® VTI Imaging Platform as described in [31]. Briefly, for every cell a binary image mask was created from the Hoechst 33342 staining signal

• with Hoechst 33342. To quantify the amount of cell-associated nanoparticles, images were analysed by using Target Activation V4 assay [56]. For every cell, a binary image mask was created from the Hoechst 33342 staining signal to define the region of interest, marking the nucleus. In the second channel

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

• ). The exposure to Ag NPs (i.e., either ALI or submerged) did not alter the morphology shown by staining F-actin with phalloidin rhodamine (red), nor could any DNA alterations be observed as visualized with DAPI (blue). Minimum intensity projections of z-stack phase contrast images revealed NPs either

• layer of the triple cell co-culture model, which is a similar pattern as we have observed for Ag NPs exposed to cells at the ALI [44]. To reduce misinterpretation due to staining artefacts [48], samples were treated with uranyl acetate only, without lead citrate. Cytotoxicity As described in [44], we

• and then treated with 0.1 M glycine in PBS for 10 min. Before staining, the cells were permeabilised with 0.2% Triton X-100 in PBS for 15 min at room temperature. The cytoskeleton (i.e., F-actin-filaments of all cells) was stained with rhodamine phalloidin 1:100 (R-415; Molecular Probes, Life

PEGylated versus non-PEGylated magnetic nanoparticles as camptothecin delivery system

• Paula M. Castillo,
• Mario de la Mata,
• Maria F. Casula,
• José A. Sánchez-Alcázar and
• Ana P. Zaderenko

Beilstein J. Nanotechnol. 2014, 5, 1312–1319, doi:10.3762/bjnano.5.144

• for their efficiency in inducing apoptosis. Apoptosis was assessed by the occurrence of cells with nuclear condensation and fragmentation by Hoechst staining [46]. In order to compare the apoptotic activity of USM[CPT] and USM-PEG[CPT] formulations, stock solutions were prepared, and suitable amounts

Model systems for studying cell adhesion and biomimetic actin networks

• Dorothea Brüggemann,
• Johannes P. Frohnmayer and
• Joachim P. Spatz

Beilstein J. Nanotechnol. 2014, 5, 1193–1202, doi:10.3762/bjnano.5.131

• this detergent removal method for reconstituting integrin αIIbβ3 from blood platelets and α1β1 from chicken gizzard into lipid vesicles. Successful integrin incorporation into liposomes of 100 to 200 nm in diameter was confirmed by negative staining in cryoelectron microscopy (Figure 2

Molecular biology approaches in bioadhesion research

• Marcelo Rodrigues,
• Birgit Lengerer,
• Thomas Ostermann and
• Peter Ladurner

Beilstein J. Nanotechnol. 2014, 5, 983–993, doi:10.3762/bjnano.5.112

• and should not have significant similarities to other endogenous transcripts (BLAST search). The size of the probes should range between 500 and 1000 nucleotides. Shorter probes can lead to weak staining results and/or less specificity. cDNA is used as a template for a standard PCR reaction with the

• a blue precipitate when it is dephosphorylated. When added to the samples NBT/BCIP leads to a stable blue staining in cells where the anti-digoxigenin antibody is bound (Figure 4). Endogenous phosphatase activity can lead to a false-positive staining. Therefore, it is essential to inhibit

• phosphatase. (4) The former colorless substrate becomes dephosphorylated and turns blue. (5) The staining reveals the cells with target gene expression, in this case the adhesive organs. Functional analyses of an adhesion-related gene by RNAi. After the application of dsRNA the mRNA gets degraded. The lack of

Biocalcite, a multifunctional inorganic polymer: Building block for calcareous sponge spicules and bioseed for the synthesis of calcium phosphate-based bone

• Xiaohong Wang,
• Heinz C. Schröder and
• Werner E. G. Müller

Beilstein J. Nanotechnol. 2014, 5, 610–621, doi:10.3762/bjnano.5.72

• a staining procedure with alizarin red S [7]. The MAC supplement (ascorbic acid, β-glycerophosphate and dexamethasone) stimulates cellular differentiation processes. Importantly, it had been measured that this process is paralleled by an enhanced expression of the CA-II gene, suggesting its

• which the alginate/cell matrix is extruded. (C) Completed 4 mm high blocks into which the cells have been embedded into the alginate. (D) The cells retain the capacity to form crystallites, which can be visualized after staining with Alizarin Red S. Acknowledgements W. E. G. M. is a holder of an ERC

Morphological characterization of fullerene–androsterone conjugates

• Alberto Ruiz,
• Margarita Suárez,
• Nazario Martin,
• Fernando Albericio and
• Hortensia Rodríguez

Beilstein J. Nanotechnol. 2014, 5, 374–379, doi:10.3762/bjnano.5.43

• aggregates were visualized using uranyl acetate negative staining. One drop of the clear solution was transferred to a TEM grid (copper grid, 3.0 mm, 200 mesh, coated with Formvar film), together with a drop of uranyl acetate (2% water solution) for 1 min, and allowed to dry. Analysis of stained grids was

Nanoscopic surfactant behavior of the porin MspA in aqueous media

• Ayomi S. Perera,
• Hongwang Wang,
• Tej B. Shrestha,
• Deryl L. Troyer and
• Stefan H. Bossmann

Beilstein J. Nanotechnol. 2013, 4, 278–284, doi:10.3762/bjnano.4.30

• microscopy (TEM). The TEM were prepared by immersing carbon-coated 200-mesh copper grids in aqueous liposome-containing solutions, followed by counter-staining by 2% aqueous uranyl acetate solution, and overnight drying in a desiccator. The dried grids were analyzed by using a HRTEM FEI Tecnai F20 XT Field

Nanolesions induced by heavy ions in human tissues: Experimental and theoretical studies

• Marcus Bleicher,
• Lucas Burigo,
• Marco Durante,
• Maren Herrlitz,
• Michael Krämer,
• Igor Mishustin,
• Iris Müller,
• Francesco Natale,
• Igor Pshenichnov,
• Stefan Schramm,
• Gisela Taucher-Scholz and
• Cathrin Wälzlein

Beilstein J. Nanotechnol. 2012, 3, 556–563, doi:10.3762/bjnano.3.64

• . DNA-damage-induced foci of the repair factor XRCC1 (green) and γH2AX (red) are clearly visualized at the sites of ion traversal. Both proteins colocalize within each of the targeted chromo centers (blue: DAPI DNA staining). (b) Analysis of the time-dependent localization of XRCC1 and γH2AX radiation

• fibroblasts. Cells were irradiated with Au ions (energy: 8 MeV/n, linear energy transfer (LET): 13000 keV/μm; fluence: 3·106 ions/cm2) at a low angle and fixed after 1 h. H4K16ac (green) is increased at damage sites. DNA damage is shown by γH2AX staining (red). DNA is counterstained with ToPro3 (blue). From