On the pathway of cellular uptake: new insight into the interaction between the cell membrane and very small nanoparticles

• Claudia Messerschmidt,
• Daniel Hofmann,
• Anja Kroeger,
• Katharina Landfester,
• Volker Mailänder and
• Ingo Lieberwirth

Beilstein J. Nanotechnol. 2016, 7, 1296–1311, doi:10.3762/bjnano.7.121

• , threshold setting and a final watershed algorithm. Particle detection was restricted between an upper and lower cut off in particle radius and to a circularity greater than 0.7. This way only separated particles were considered for measurement. Cell culture For TEM experiments, all different cell lines were

• suspension was added to the cell culture medium, yielding the final concentration for exposure. For transmission electron microscopy and for cytotoxicity assays, cells were incubated with SiNPs for 10 min to 24 h at 37 °C and 5% CO2 before further processing. To check if the observed particle engulfment is

• York City, U.S.A.) in triplicates and incubated for 20 h. After particle load and incubation, cells were centrifuged for 10 min at 600g. Then, 10 µL of cell culture supernatant was transferred into a fresh 96-well plate, mixed with 100 µL of the LDH reaction mix and incubated for 60 min at room

High antiviral effect of TiO₂·PL–DNA nanocomposites targeted to conservative regions of (−)RNA and (+)RNA of influenza A virus in cell culture

• Asya S. Levina,
• Marina N. Repkova,
• Elena V. Bessudnova,
• Ekaterina I. Filippova,
• Natalia A. Mazurkova and
• Valentina F. Zarytova

Beilstein J. Nanotechnol. 2016, 7, 1166–1173, doi:10.3762/bjnano.7.108

• ). It was demonstrated that these nanocomposites exhibited a low toxicity and very high activity against IAV in the cell culture [18][19]. Only one site in the IAV 5 segment was used in our previous works. In this work, we examined the antiviral activity of DNA fragments in the proposed TiO2·PL–DNA

• inhibit the reproduction of IAV in cell culture. TiO2 nanoparticles (of ≈5 nm in diameter) are known to penetrate through cell membrane [23]. It was clearly demonstrated in our previous work [17][18] that they are good vehicles to transport DNA fragments into cells. There are literature data that show

Improved biocompatibility and efficient labeling of neural stem cells with poly(L-lysine)-coated maghemite nanoparticles

• Igor M. Pongrac,
• Marina Dobrivojević,
• Lada Brkić Ahmed,
• Michal Babič,
• Miroslav Šlouf,
• Daniel Horák and
• Srećko Gajović

Beilstein J. Nanotechnol. 2016, 7, 926–936, doi:10.3762/bjnano.7.84

• of both types of the nanoparticles occurred via macropinocytosis as confirmed by TEM. Recently, PLL-γ-Fe2O3 agglomeration properties were studied in biological cell culture media with or without common serum protein, which showed the increase of size and negative ζ-potential in comparison to

• , Great Britain). The agglomeration properties and the surface charge properties of PLL-γ-Fe2O3 nanoparticles in biological cell culture medium with and without addition of common serum protein were previously described [26]. The crystal structure of both types of nanoparticles was investigated using the

• . All experiments were carried out in accordance with the EU Directive 2010/63/EU on the protection of animals used for scientific purposes. Neural stem cell culture Neural stem cells (NSCs) were isolated from pregnant female mice as previously described [28][29]. Briefly, at gestation day 14.5, embryos

Novel roles for well-known players: from tobacco mosaic virus pests to enzymatically active assemblies

• Claudia Koch,
• Fabian J. Eber,
• Carlos Azucena,
• Alexander Förste,
• Stefan Walheim,
• Thomas Schimmel,
• Alexander M. Bittner,
• Holger Jeske,
• Hartmut Gliemann,
• Sabine Eiben,
• Fania C. Geiger and
• Christina Wege

Beilstein J. Nanotechnol. 2016, 7, 613–629, doi:10.3762/bjnano.7.54

• , energy conversion, plasmonics or magnetic resonance imaging [82][92][100][114][115][116][117][118][119][120][121], and as carrier rods for effector peptides for distinct purposes from affinity binding, intravital targeting up to cell-culture supports [105][111][117][122][123][124], or as antigens for

Functional fusion of living systems with synthetic electrode interfaces

• Oskar Staufer,
• Sebastian Weber,
• C. Peter Bengtson,
• Hilmar Bading,
• Joachim P. Spatz and
• Amin Rustom

Beilstein J. Nanotechnol. 2016, 7, 296–301, doi:10.3762/bjnano.7.27

• experimental information. Acknowledgements We would like to thank It4ip Belgium for providing us with track-etched PC filter membranes with custom parameters, Sigrid Riese for cell culture and experimental support, Andrea Hellwig for support with SEM and TEM measurements, and Günther Meinusch for help with

Surface coating affects behavior of metallic nanoparticles in a biological environment

• Darija Domazet Jurašin,
• Marija Ćurlin,
• Ivona Capjak,
• Tea Crnković,
• Marija Lovrić,
• Michal Babič,
• Daniel Horák,
• Ivana Vinković Vrček and
• Srećko Gajović

Beilstein J. Nanotechnol. 2016, 7, 246–262, doi:10.3762/bjnano.7.23

• . Three dispersion media were investigated: ultrapure water (UW), biological cell culture medium without addition of protein (BM), and BM supplemented with common serum protein (BMP). The obtained results showed that different coating agents on AgNPs and SPIONs produced different stabilities in the same

• properties and behavior in a biological environment. Despite a considerable number of studies on the colloidal stability of AgNPs and SPIONs in cell culture media, in natural water, or in the formulation of consumer products [2][7][8][9][10][11][12][13][14], general conclusions and a clear understanding of

• albumin (BSAAgNPs), Brij 35 (BrijAgNP) and Tween 20 (TweenAgNP). The SPIONs were prepared as uncoated γ-Fe2O3 NPs (UNSPIONs), and coated with D-mannose (MANSPIONs) or poly(L-lysine) (PLLSPIONs). Three media for NP dispersion were investigated: ultrapure water (UW), biological cell culture medium without

Synthesis, characterization and in vitro biocompatibility study of Au/TMC/Fe₃O₄ nanocomposites as a promising, nontoxic system for biomedical applications

• Hanieh Shirazi,
• Maryam Daneshpour,
• Soheila Kashanian and
• Kobra Omidfar

Beilstein J. Nanotechnol. 2015, 6, 1677–1689, doi:10.3762/bjnano.6.170

• magnetic and optoelectronic properties. Experimental Reagents and solutions Chloroauric acid (HAuCl4), sodium dodecyl sulfate (SDS), sodium azide, low molecular weight chitosan, MTT reagent, RPMI1640 cell culture medium, fetal bovine serum (FBS), and dialysis tubing with a molecular cutoff of 12000 Da were

Influence of surface chemical properties on the toxicity of engineered zinc oxide nanoparticles to embryonic zebrafish

• Zitao Zhou,
• Jino Son,
• Bryan Harper,
• Zheng Zhou and
• Stacey Harper

Beilstein J. Nanotechnol. 2015, 6, 1568–1579, doi:10.3762/bjnano.6.160

• have been noted to contribute to lower viability in cell culture studies with A549 and HT29 cells [30]. Similar morphology effects on toxicity have been observed in studies of manganese oxide, where the sharp points and edges were found to generate more ROS than smooth surfaces [44]. We tested this

PLGA nanoparticles as a platform for vitamin D-based cancer therapy

• Maria J. Ramalho,
• Joana A. Loureiro,
• Bárbara Gomes,
• Manuela F. Frasco,
• Manuel A. N. Coelho and
• M. Carmo Pereira

Beilstein J. Nanotechnol. 2015, 6, 1306–1318, doi:10.3762/bjnano.6.135

• was tested with concentrations of PLGA in the range of 0.1 µg mL−1 to 50 µg mL−1. The efficacy of calcitriol-loaded NPs in comparison to free calcitriol was also evaluated. Due to its known short half-life in the cell culture medium, an assay with S2-013 cells was performed for 48 h to compare cell

• effect (Table 3). Still, it is important to highlight that due to the short calcitriol half-life in the cell culture medium, the presented free drug concentrations were added daily, unlike entrapped calcitriol, which was loaded in the NPs only once at time t = 0. Thus, despite that the NPs themselves

• equilibrium between the NPs and release medium occurred. The in vitro cytotoxic studies proved that unloaded PLGA NPs are biocompatible and revealed the toxicity effect of calcitriol against human pancreatic and lung cells. Due to the short calcitriol half-life in the cell culture medium, daily renewal was

Synthesis, characterization and in vitro effects of 7 nm alloyed silver–gold nanoparticles

• Simon Ristig,
• Svitlana Chernousova,
• Wolfgang Meyer-Zaika and
• Matthias Epple

Beilstein J. Nanotechnol. 2015, 6, 1212–1220, doi:10.3762/bjnano.6.124

• can be verified. Cell culture experiments To examine the cytotoxicity with regards to the molar fraction of silver in the nanoparticles, HeLa cells and human mesenchymal stem cells were incubated with nanoalloys of nine different compositions and also with pure gold and pure silver nanoparticles. In

• nanoparticles themselves [9][40][41], it is important to separate the toxic effects of the nanoparticles and unreacted material from the synthesis [11][42]. As some reported cell culture experiments with alloyed silver–gold nanoparticles were conducted without purification of the dispersions [19][43][44], it

• volume of 3.5 mL were used. Atomic absorption spectroscopy (AAS) was carried out with a Thermo Electron M-Series spectrometer with a graphite tube furnace according to DIN EN ISO/IEC 17025:2005 after dissolving the particles in aqua regia. Cell culture HeLa cells (human transformed cervix epithelial

Tattoo ink nanoparticles in skin tissue and fibroblasts

• Colin A. Grant,
• Peter C. Twigg,
• Richard Baker and
• Desmond J. Tobin

Beilstein J. Nanotechnol. 2015, 6, 1183–1191, doi:10.3762/bjnano.6.120

• agglomeration behaviour of the particles will be strongly influenced by the in vivo conditions in tattooing, or the in vitro conditions in cell culture. Agglomeration due to electrochemical processes can reduce the effective number of particles by orders of magnitude and this will have a profound effect on how

Influence of gold, silver and gold–silver alloy nanoparticles on germ cell function and embryo development

• Ulrike Taylor,
• Daniela Tiedemann,
• Christoph Rehbock,
• Wilfried A. Kues,
• Stephan Barcikowski and
• Detlef Rath

Beilstein J. Nanotechnol. 2015, 6, 651–664, doi:10.3762/bjnano.6.66

• traditionally used in somatic cell culture. Table 1 gives examples of common reprotoxicological in vitro tests and their predictive values. However, despite its obvious importance, reprotoxicological testing of nanoparticles has so far been frequently neglected. While in the last two years approximately 1000

Novel ZnO:Ag nanocomposites induce significant oxidative stress in human fibroblast malignant melanoma (Ht144) cells

• Syeda Arooj,
• Samina Nazir,
• Akhtar Nadhman,
• Nafees Ahmad,
• Bakhtiar Muhammad,
• Ishaq Ahmad,
• Kehkashan Mazhar and
• Rashda Abbasi

Beilstein J. Nanotechnol. 2015, 6, 570–582, doi:10.3762/bjnano.6.59

• of the nanocomposites were calculated by the Kubelka–Munk function F(R) by using the following equation [30]. where F(R) is the Kubelka–Munk function; R represents the absolute value of reflectance. Cell lines and cell culture Human malignant melanoma (HT144, ATCC HTB-63) cells were cultured in RPMI

• in de-ionized water and dispersed by sonication on sonifier cell disrupter (40 W, 25 kHz) for 30 min in a water bath. The nanoparticle serial dilutions were prepared as required by using the appropriate cell culture medium. To screen the photo-oxidation-mediated effects of the NPs, briefly, 1.5 × 105

Pulmonary surfactant augments cytotoxicity of silica nanoparticles: Studies on an in vitro air–blood barrier model

• Jennifer Y. Kasper,
• Lisa Feiden,
• Maria I. Hermanns,
• Christoph Bantz,
• Michael Maskos,
• Ronald E. Unger and
• C. James Kirkpatrick

Beilstein J. Nanotechnol. 2015, 6, 517–528, doi:10.3762/bjnano.6.54

• nanoparticles. Particle size was determined using the method of dynamic light scattering (DLS). Thus, the reported sizes are z-weighted mean values of the hydrodynamic diameter. Particle diameters were measured in cell culture medium (RPMI 1640) and, for reference, in water (containing 2 mmol/L sodium bromide

• change in particle size could be detected for any of the particle types in water as well as in RPMI 1640 cell culture medium. Furthermore, no significantly different particle sizes were measured for any of the different surface modifications. As the particles exhibit comparable sizes and an identical

• nanometers in diameter were found in cell culture medium (data not shown). However, the analysis of the pure Alveofact® dispersion revealed that agglomerates of the same size were already present without nanoparticles. This leads to the conclusion that the standard procedure proposed by the manufacturer for

A surface acoustic wave-driven micropump for particle uptake investigation under physiological flow conditions in very small volumes

• Florian G. Strobl,
• Dominik Breyer,
• Phillip Link,
• Adriano A. Torrano,
• Christoph Bräuchle,
• Matthias F. Schneider and
• Achim Wixforth

Beilstein J. Nanotechnol. 2015, 6, 414–419, doi:10.3762/bjnano.6.41

• uptake of nanoparticles. Here, we present a versatile microfluidic device based on acoustic streaming induced by surface acoustic waves (SAWs). The device offers a convenient method for introducing fluid motion in standard cell culture chambers and for mimicking capillary blood flow. We show that shear

• to the aforementioned problems is to perform experiments under realistic flow conditions. In the following, we introduce a novel device for inducing high shear rates at the bottom of an arbitrary cell culture chamber. The device is based on SAW-driven acoustic streaming. Many different applications

Hematopoietic and mesenchymal stem cells: polymeric nanoparticle uptake and lineage differentiation

• Ivonne Brüstle,
• Thomas Simmet,
• Gerd Ulrich Nienhaus,
• Katharina Landfester and
• Volker Mailänder

Beilstein J. Nanotechnol. 2015, 6, 383–395, doi:10.3762/bjnano.6.38

• differentiation was induced using NH Osteo Diff Medium/NH Chrondro Diff Medium (Miltenyi Biotec Bergisch Gladbach, Germany); the medium was changed twice a week. Analysis was performed after 24 days of cell culture. Before starting the differentiation, all cells were incubated with 300 µg/mL nanoparticles for 24

• the d0 sample as 1. The relative expression was used if d0 samples did not show expression of the used marker gene and was calculated with the ΔCT method, setting the expression in the sample without particles to 1. HTRF Assay The levels of IL-8 in the cell culture supernatant were quantitatively

• performed after 5 days of cultivation with a HTRF assay by analyzing the cell culture supernatant. p > 0.05 ns, p < 0.001 ***. Cytochemical staining to determine the differentiation of hMSCs incubated with different nanoparticles (300 µg/mL nanoparticles, incubation time 24 h, before inducing

Comparative evaluation of the impact on endothelial cells induced by different nanoparticle structures and functionalization

• Lisa Landgraf,
• Ines Müller,
• Peter Ernst,
• Miriam Schäfer,
• Christina Rosman,
• Isabel Schick,
• Oskar Köhler,
• Hartmut Oehring,
• Vladimir V. Breus,
• Thomas Basché,
• Carsten Sönnichsen,
• Wolfgang Tremel and
• Ingrid Hilger

Beilstein J. Nanotechnol. 2015, 6, 300–312, doi:10.3762/bjnano.6.28

• according to the experiment and the cell line. Cell culture The mouse endothelial (SVEC4-10) cell line was obtained from American Type Culture Collection (LGC Standards GmbH, Germany). Cells were grown in Dulbecco´s modified eagle´s medium (DMEM with Glutamax; Invitrogen, Germany) supplemented with 10% heat

The effect of surface charge on nonspecific uptake and cytotoxicity of CdSe/ZnS core/shell quantum dots

• Vladimir V. Breus,
• Anna Pietuch,
• Marco Tarantola,
• Thomas Basché and
• Andreas Janshoff

Beilstein J. Nanotechnol. 2015, 6, 281–292, doi:10.3762/bjnano.6.26

• (Figure 3a). The growth of CA–QDs aggregates over time could be caused by opsonization (e.g., the adsorption of proteins from the cell culture medium on the nanoparticles). Serum-induced aggregation was reported earlier for CdSe/ZnS QDs with cationic charge [19]. Some very large fluorescent aggregates can

• early stages. This work highlights the importance of the combined use of ECIS and MTS (or other biochemical) assays for the characterization of nanoparticle cytotoxicity. Experimental Cell culture MDCKII cells were maintained in Earle’s minimum essential medium supplemented with 4 mM glutamine, 100 g/mL

• , followed by trypsinization and centrifugation at 110g. Counting was carried out using a Neubauer chamber, and viability was determined using trypan blue exclusion. For fluorescence microscopy measurements, the cells were grown in 2 mL of cell culture medium in a ibiTreat µ-Dish (Ibidi, Martinsried, Germany

Release behaviour and toxicity evaluation of levodopa from carboxylated single-walled carbon nanotubes

• Julia M. Tan,
• Jhi Biau Foo,
• Sharida Fakurazi and
• Mohd Zobir Hussein

Beilstein J. Nanotechnol. 2015, 6, 243–253, doi:10.3762/bjnano.6.23

• Mytilineou et al. [30], where the LD exposed to mesencephalic cultures at 72 h of treatment was associated with a dose-dependent reduction in cell survival. The authors attributed the potential toxicity of LD to the decreased antioxidant capacity in the limited environment of the cell culture that makes the

Tailoring the ligand shell for the control of cellular uptake and optical properties of nanocrystals

• Johannes Ostermann,
• Christian Schmidtke,
• Christopher Wolter,
• Jan-Philip Merkl,
• Hauke Kloust and
• Horst Weller

Beilstein J. Nanotechnol. 2015, 6, 232–242, doi:10.3762/bjnano.6.22

• copolymers. Tested samples and the observed interaction with A549 cells. Supporting Information Supporting Information File 96: Additional information about the conducted cell culture experiments available. Acknowledgements This work was supported by the Deutsche Forschungsgemeinschaft (DFG, SPP 1313), the

Caveolin-1 and CDC42 mediated endocytosis of silica-coated iron oxide nanoparticles in HeLa cells

• Nils Bohmer and
• Andreas Jordan

Beilstein J. Nanotechnol. 2015, 6, 167–176, doi:10.3762/bjnano.6.16

• , the fluorescent dye Alexa Fluor® 555 was embedded into their shells. Further details of synthesis and characterization have been described elsewhere [29][30]. Cell culture HeLa cells (human cervix carcinoma) were provided by the group of Professor Haucke (Freie Universität, Berlin, Germany). They were

• 70–80% confluence. Before every single experiment, nanoparticles were prewarmed to 37 °C and treated with ultrasound for 10 min to avoid sedimentation and aggregation. SPIONs were diluted with cell culture media to a concentration of 50 µg Fe/mL while SCIONs were diluted to 5 µg/mL. The concentration

• of the SCIONs was chosen after preliminary experiments. It could be shown that 5 µg Fe/mL provides the lowest background fluorescence combined with a good intracellular signal. In both setups the cell culture media contains SCIONs in excess to the internalization rate of the cells. Afterward cells

Increasing throughput of AFM-based single cell adhesion measurements through multisubstrate surfaces

• Miao Yu,
• Nico Strohmeyer,
• Jinghe Wang,
• Daniel J. Müller and
• Jonne Helenius

Beilstein J. Nanotechnol. 2015, 6, 157–166, doi:10.3762/bjnano.6.15

• hydrophilic during plasma treatment, and thus, liquid coating drops spread over the masks. Storing the PDMS in air will restore the hydrophobicity of the surface. Cell culture PC3 cells were maintained in RPMI-1640-supplemented (Gibco-Life technologies) 1 mM sodium pyruvate; HeLa cells (Kyoto) and mouse

The distribution and degradation of radiolabeled superparamagnetic iron oxide nanoparticles and quantum dots in mice

• Denise Bargheer,
• Artur Giemsa,
• Barbara Freund,
• Markus Heine,
• Christian Waurisch,
• Gordon M. Stachowski,
• Stephen G. Hickey,
• Alexander Eychmüller,
• Jörg Heeren and
• Peter Nielsen

Beilstein J. Nanotechnol. 2015, 6, 111–123, doi:10.3762/bjnano.6.11

• various Qdots in cell culture [7][8][9][10][11][12]. However, when dosing was applied under physiological conditions in mice and rats, no abnormal behavior or tissue damage was observed over the period of months after systemic administration of Qdots [13][14][15]. Therefore, the relevancy of the in vitro

• cellular internalization and processing of polymer-coated Qdots and SPIOs was gained by cell culture. For comparison of the in vivo data, these studies were made with J774 cells (a murine macrophage cell line). The cells were incubated with polymer-coated Qdots for 2 h, then the nanoparticles were removed

• µm thicknesses were cut from frozen specimens embedded in an optimum cutting temperature formulation solution (Sakura Finetek Europe). Cryosections were mounted on a slide (Superfrost/Plus, Glaswarenfabrik Karl Hecht KG, Sondheim, Germany) and dried overnight before immunostaining. Cell culture J774

The fate of a designed protein corona on nanoparticles in vitro and in vivo

• Denise Bargheer,
• Julius Nielsen,
• Gabriella Gébel,
• Markus Heine,
• Sunhild C. Salmen,
• Roland Stauber,
• Horst Weller,
• Joerg Heeren and
• Peter Nielsen

Beilstein J. Nanotechnol. 2015, 6, 36–46, doi:10.3762/bjnano.6.5

• transferrin receptor binding on the surface of cells [23]. It should be noted that most of the mentioned work was performed in in vitro experiments using partly also cell culture models. To date, much less information is available on the consequences of protein adsorption on nanoparticles in vivo. This is not

• charged SPIOs had no preformed transferrin corona but must have been also coated by plasma proteins before liver uptake took place implicating a similar biodistribution. This confirms the stability of a corona in a recent study by Wang et al. using a cell culture model [22]. Conclusion This pilot study

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

• ]). Cell culture In our studies, we used epithelial MDCK (type II) cells and performed cell culture as described in [20]. Cells were cultured in Earle’s minimum essential medium supplemented with glutamine (4 mM), penicillin and streptomycin (100 µg/mL for both), fetal calf serum (10% v/v) and stored in an

• optical dark field microscopy measurements. Finally, 10 μL of the cell suspension (≈6000 cells) were seeded and samples were kept under cell culture conditions. Live cell imaging Optical dark field microscopy was performed after 4, 24, 48, and 72 h while maintaining 37 °C and a 5% CO2 environment. In

• order to improve contrast for imaging, the cell culture medium was replaced by a version without a phenol red pH indicator or any nutrients. As soon as imaging was finished, the medium was changed back to the full culture medium and the samples were kept in a cell culture incubator. Electric cell