Toxicity and safety study of silver and gold nanoparticles functionalized with cysteine and glutathione

• Barbara Pem,
• Igor M. Pongrac,
• Lea Ulm,
• Ivan Pavičić,
• Valerije Vrček,
• Darija Domazet Jurašin,
• Marija Ljubojević,
• Adela Krivohlavek and
• Ivana Vinković Vrček

Beilstein J. Nanotechnol. 2019, 10, 1802–1817, doi:10.3762/bjnano.10.175

• were regularly tested for absence of mycoplasma by means of a direct DNA dye test [85]. When the cells reached 80% confluence, the culture medium was removed with a pipette; the cells were washed once with sterile phosphate buffer saline (PBS), detached from the flask by adding trypsin/EDTA (0.25

• untreated cells were negative controls. After treatment, the plates were centrifuged at 1500g for 15 min. Supernatants containing dead cells were collected from 12-well plates in 1.5 mL Eppendorf tubes. Live cells were detached from the wells by adding 0.05 % GibcoTM trypsin/EDTA (Thermo Fisher Scientific

Enhanced inhibition of influenza virus infection by peptide–noble-metal nanoparticle conjugates

• Zaid K. Alghrair,
• David G. Fernig and
• Bahram Ebrahimi

Beilstein J. Nanotechnol. 2019, 10, 1038–1047, doi:10.3762/bjnano.10.104

• streptomycin (Gibco, Life Technologies, UK) and incubated in a humidified environment at 37 °C under 5% (v/v) CO2 atmosphere. Cells were detached with 0.05% (w/v) trypsin in the chelating agent, 1× Versene-EDTA (Gibco, Life Technologies, UK) and plated at a dilution of 1:4. Preparation of influenza virus stock

• DMEM. Cells were incubated with virus for 1 h at 37 °C on a rocking platform. Virus-containing medium was removed and the cell monolayer washed with 2 × 5 mL DMEM, then 5 mL N-acetyl trypsin (Sigma-Aldrich, Merck, UK), 2.5 µg/mL in DMEM, was added and incubated for 24–48 hours at 37 °C until a

• % (w/v) NaHCO2, 1.4 mL 1 M HEPES, 1.4 mL (1% (v/v) 100 U/mL penicillin and 1% (v/v) 100 µg/mL streptomycin), 44.8 mL H2O and 5 µL N-acetyl trypsin) to give a final 1% (w/v) agarose mixture. After 1 h of incubation of the cells with the virus, the supernatant was removed from the plates and overlaid

Serum type and concentration both affect the protein-corona composition of PLGA nanoparticles

• Katrin Partikel,
• Robin Korte,
• Dennis Mulac,
• Hans-Ulrich Humpf and
• Klaus Langer

Beilstein J. Nanotechnol. 2019, 10, 1002–1015, doi:10.3762/bjnano.10.101

• (Karlsruhe, Germany). The dye Coomassie Brilliant Blue G-250 was purchased from VWR Life science AMRESCO (Solon, Ohio). Bovine serum albumin (BSA), DL-dithiothreitol (DTT), and iodoacetamide (IAA) were obtained from Sigma-Aldrich (Steinheim, Germany). Trypsin (sequencing grade) for protein digestion was

• tryptic in-solution digestion of proteins, the samples were diluted to a total volume of 1000 µL with ultrapure water to a final concentration of 0.6 M urea to maintain the activity of trypsin. Next, 10 µL of ice-cooled trypsin solution (200 ng/µL) was added to the diluted samples and digestion was

• entries) using the PEAKS de novo algorithm and the enhanced target-decoy method (“decoy fusion”) for false discovery rate (FDR) estimation and result validation [40][41]. Search parameters were: (a) trypsin as specific enzyme, three missed cleavage allowed; (b) fixed modification: carbamidomethylation of

Characterization and influence of hydroxyapatite nanopowders on living cells

• Przemyslaw Oberbek,
• Tomasz Bolek,
• Adrian Chlanda,
• Seishiro Hirano,
• Sylwia Kusnieruk,
• Julia Rogowska-Tylman,
• Ganna Nechyporenko,
• Viktor Zinchenko,
• Wojciech Swieszkowski and
• Tomasz Puzyn

Beilstein J. Nanotechnol. 2018, 9, 3079–3094, doi:10.3762/bjnano.9.286

• Corporation). All culture media were supplemented with 10% heat-inactivated fetal bovine serum (FBS, HyClone™), 100 μg/mL streptomycin and 100 μg/mL penicillin. The cells were cultured at 37 °C in an incubator with a humidified atmosphere containing 5% CO2. For the passage procedure, 0.05% Trypsin–EDTA with

Comparative biological effects of spherical noble metal nanoparticles (Rh, Pd, Ag, Pt, Au) with 4–8 nm diameter

• Alexander Rostek,
• Marina Breisch,
• Kevin Pappert,
• Kateryna Loza,
• Marc Heggen,
• Manfred Köller,
• Christina Sengstock and
• Matthias Epple

Beilstein J. Nanotechnol. 2018, 9, 2763–2774, doi:10.3762/bjnano.9.258

• by the addition of 0.2 mL cm−2 0.25% trypsin/0.05% ethylenediaminetetraacetic acid (EDTA, Sigma-Aldrich, Taufkirchen, Germany) for 5 min at 37 °C. Subsequently, the hMSC cells were collected and washed twice with RPMI1640/10% FCS. Subconfluent hMSC cells were seeded in 24-well cell culture plates

Non-agglomerated silicon–organic nanoparticles and their nanocomplexes with oligonucleotides: synthesis and properties

• Asya S. Levina,
• Marina N. Repkova,
• Nadezhda V. Shikina,
• Zinfer R. Ismagilov,
• Svetlana A. Yashnik,
• Dmitrii V. Semenov,
• Yulia I. Savinovskaya,
• Natalia A. Mazurkova,
• Inna A. Pyshnaya and
• Valentina F. Zarytova

Beilstein J. Nanotechnol. 2018, 9, 2516–2525, doi:10.3762/bjnano.9.234

• can effectively and selectively interact with RNA targets in the cell culture. Experimental Materials and methods All chemicals were obtained from commercial suppliers as follows: aminiopropyltriethoxysilane (APTES) and fluoresceinisothiocyanate (FITC), trypsin, penicillin-streptomycin, and L

• -glutamine (Sigma-Aldrich, USA); RPMI-1640 medium; antibiotics (BioloT, Russia); fetal calf serum (Gibco, USA). Сhicken erythrocytes, MDCK cells, and influenza A virus strain A/chicken/Kurgan/05/2005 (H5N1) were from FBRI Vector, Russia. Trypsin (1 mg/mL) and penicillin-streptomycin (100 U/mL) were stored at

• [22]. The influenza A virus (IAV) strains and MDCK cells were prepared as in [22]. The cells at ≈80% confluence were initially infected with A/chicken/Kurgan/05/2005 virus (H5N1), which was added in each well in RPMI-1640 medium (100 μL) containing trypsin (2 μg/mL) at a MOI of 0.1. The control sample

The structural and chemical basis of temporary adhesion in the sea star Asterina gibbosa

• Birgit Lengerer,
• Marie Bonneel,
• Mathilde Lefevre,
• Elise Hennebert,
• Philippe Leclère,
• Emmanuel Gosselin,
• Peter Ladurner and
• Patrick Flammang

Beilstein J. Nanotechnol. 2018, 9, 2071–2086, doi:10.3762/bjnano.9.196

• and rehydration, antigen retrieval with a solution of 0.05% trypsin and 0.1% CaCl2 was performed for 15 min on 37 °C. Footprints were collected on microscope glass slides and fixed in 4% PFA in PBS overnight at room temperature. All samples were blocked in PBS containing 3% (w/v) bovine serum albumin

Biomimetic and biodegradable cellulose acetate scaffolds loaded with dexamethasone for bone implants

• Aikaterini-Rafailia Tsiapla,
• Varvara Karagkiozaki,
• Veroniki Bakola,
• Foteini Pappa,
• Panagiota Gkertsiou,
• Eleni Pavlidou and
• Stergios Logothetidis

Beilstein J. Nanotechnol. 2018, 9, 1986–1994, doi:10.3762/bjnano.9.189

• through electrospinning, in order to prevent the inflammation that can occur after a total hip replacement surgery. Experimental Materials and methods Materials Cellulose acetate (CA, Mw = 30,000 g/mol), dexamethasone (≥97%), acetone (≥99.8%), trypsin and methylene blue were all purchased from Sigma

Enzymatically promoted release of organic molecules linked to magnetic nanoparticles

• Chiara Lambruschini,
• Silvia Villa,
• Luca Banfi,
• Fabio Canepa,
• Fabio Morana,
• Annalisa Relini,
• Paola Riani,
• Renata Riva and
• Fulvio Silvetti

Beilstein J. Nanotechnol. 2018, 9, 986–999, doi:10.3762/bjnano.9.92

• fluorophore from the tripeptide. In order to check the affinity of our peptide, and to select the correct amount of enzyme to be used, we carried out some experiments with model compound 7, using trypsin and plasmin as proteases. Trypsin, like plasmin, has a preference for lysine (or arginine) as the scissile

• pyrenylmethylamine from 50 nmol of 7 in 72 h. The conversion was already 88% after 24 h. Trypsin displayed a similar behaviour. The units for this enzyme were not provided, but comparing the rates, we established that 170 mg of trypsin had the same catalytic efficiency as 1 U of plasmin. Thus, reaction on 50 nmol of

• 7 was complete in 48 h using 4.6 μg of trypsin. In both cases, the kinetics was found to be first order with respect to the substrate. Since the aim of our work was mainly to check the compatibility of the nanoparticles with the enzymatic reaction, the more available trypsin was used in the

Green synthesis of fluorescent carbon dots from spices for in vitro imaging and tumour cell growth inhibition

• Nagamalai Vasimalai,
• Vânia Vilas-Boas,
• Juan Gallo,
• María de Fátima Cerqueira,
• Mario Menéndez-Miranda,
• José Manuel Costa-Fernández,
• Lorena Diéguez,
• Begoña Espiña and
• María Teresa Fernández-Argüelles

Beilstein J. Nanotechnol. 2018, 9, 530–544, doi:10.3762/bjnano.9.51

• (DMEM) with high glucose content and all other chemicals were purchased from Sigma-Aldrich (Spain). Ultrapure water (18.2 MΩ·cm at 25 °C, Millipore USA) was used throughout the experiments. Dulbecco’s modified Eagle’s medium with nutrient mixture F-12 (DMEM/F-12) and GlutaMAX-I™, trypsin 0.25%–EDTA

Nanoparticle delivery to metastatic breast cancer cells by nanoengineered mesenchymal stem cells

• Liga Saulite,
• Karlis Pleiko,
• Ineta Popena,
• Dominyka Dapkute,
• Ricardas Rotomskis and
• Una Riekstina

Beilstein J. Nanotechnol. 2018, 9, 321–332, doi:10.3762/bjnano.9.32

• up to 90% confluence in complete cell culture medium in a humidified chamber at 37 °C with 5% CO2. For passaging, the cells were trypsinised using 0.25% trypsin–EDTA solution. All cell reagents were purchased from Sigma-Aldrich, St. Louis, MO, USA. Establishment of 3D cell culture model A poly(2

• polyHEMA-coated plates for co-culture with 2.5 × 104 MCF7 or MDA-MB-231 cells (2:1) in complete medium [52][53]. After 24 h, supernatants containing floating spheroids were aspirated and centrifuged at 250 g for 5 min. The pellet was suspended in 0.25% trypsin–EDTA for 5 min at 37 °C to ensure the

Low uptake of silica nanoparticles in Caco-2 intestinal epithelial barriers

• Dong Ye,
• Mattia Bramini,
• Delyan R. Hristov,
• Sha Wan,
• Anna Salvati,
• Christoffer Åberg and
• Kenneth A. Dawson

Beilstein J. Nanotechnol. 2017, 8, 1396–1406, doi:10.3762/bjnano.8.141

• cytometry. After exposure to 100 µg/mL nanoparticles dispersed in the absence and presence of 10 % FBS, the dispersions were removed from each well and cells were rinsed twice with fresh phosphate buffered saline (PBS). Then, 1 mL 0.1% trypsin-ethylenediaminetetraacetic acid (EDTA) solution was added and

• light microscope. The detached cells were then collected from each well and the same volume of complete medium added to inhibit the trypsin. Cell pellets were harvested by centrifugation at 1,500 RPM for 3 min and resuspended in fresh PBS. In order to fix the cells, 4% formalin solution (Sigma-Aldrich

Carbon nanomaterials sensitize prostate cancer cells to docetaxel and mitomycin C via induction of apoptosis and inhibition of proliferation

• Kati Erdmann,
• Jessica Ringel,
• Silke Hampel,
• Manfred P. Wirth and
• Susanne Fuessel

Beilstein J. Nanotechnol. 2017, 8, 1307–1317, doi:10.3762/bjnano.8.132

• rate For examination of cell proliferation, cell colony formation and apoptosis cells were seeded into 6-well plates and incubated as indicated above. Seventy two hours after the end of treatment cells were harvested by trypsin/EDTA treatment and submitted to the aforementioned assays. Cell

Characterization of ferrite nanoparticles for preparation of biocomposites

• Urszula Klekotka,
• Magdalena Rogowska,
• Dariusz Satuła and
• Beata Kalska-Szostko

Beilstein J. Nanotechnol. 2017, 8, 1257–1265, doi:10.3762/bjnano.8.127

• with a bioparticle was studied. In subsequent steps, the nanoparticles were immobilized with enzymes such as albumin, glucose oxidase, lipase and trypsin as a test bioparticles. The characterization of the nanoparticles was acheived by transmission electron microscopy, X-ray diffraction, energy

• ; magnetic nanoparticles; surface immobilization; trypsin; X-ray diffraction; Introduction Nanoparticles are important ingredients in the fabrication of biocomposites, and therefore, the surface functionalization of nanoparticles attracts great interest among scientists [1][2]. Tests that aim at the

• , we have used nanoparticles with or without attached glutaraldehyde that served as a linker between the nanoparticle and enzymes and gives more space for interaction. The enzymes tested in this paper were: albumin, glucose oxidase, lipase, and trypsin. This study is a continuation of our previous

Evaluation of quantum dot conjugated antibodies for immunofluorescent labelling of cellular targets

• Jennifer E. Francis,
• David Mason and
• Raphaël Lévy

Beilstein J. Nanotechnol. 2017, 8, 1238–1249, doi:10.3762/bjnano.8.125

• 75 cm2 flask at 37 °C with 5% CO2, minimum essential media (MEM, Life Technologies, UK) supplemented with 10% (v/v) foetal calf serum (FCS), and 1% non-essential amino acids (NEAA). Cells were split 1,000,000 cells/mL when ≥80% confluent with trypsin-EDTA. Rat mammary (Rama) 27 fibroblasts were

• previously [40]. Cells were split 1:8 when ≥60% confluent with trypsin-EDTA. A stable cell line TC7 3xGFP (expressing tubulin-GFP) was cultured in a 75 cm2 flask at 37 °C with 5% CO2, MEM (Life Technologies, UK) supplemented with 10% (v/v) FCS, 1% NEAA, and genetitin (Sigma-Aldrich, UK), as described

• previously [41]. Cells were split 1:15 when ≥80% confluent with trypsin-EDTA. Transfection HeLa cells were seeded onto 16 mm glass coverslips (100,000 cells/mL) in a 12-well plate and transfected with pG-EGFP-A (soluble GFP) or pG-EGFP-HIF2α (EGFP-HIF2α) using FuGENE6 transfection reagent (Roche Limited, UK

Nano-engineered skin mesenchymal stem cells: potential vehicles for tumour-targeted quantum-dot delivery

• Liga Saulite,
• Dominyka Dapkute,
• Karlis Pleiko,
• Ineta Popena,
• Simona Steponkiene,
• Ricardas Rotomskis and
• Una Riekstina

Beilstein J. Nanotechnol. 2017, 8, 1218–1230, doi:10.3762/bjnano.8.123

• streptomycin) (all from Sigma-Aldrich, USA). Cell suspensions were transferred into 25 cm2 tissue culture flasks and grown until reaching 80% confluence in a humidified chamber at 37 °C with 5% CO2. Cells were trypsinized with 0.25% trypsin–EDTA solution (Sigma-Aldrich, USA). Cells at passages 2 to 5 were then

Surface-enhanced Raman spectroscopy of cell lysates mixed with silver nanoparticles for tumor classification

• Mohamed Hassoun,
• Iwan W.Schie,
• Tatiana Tolstik,
• Sarmiza E. Stanca,
• Christoph Krafft and
• Juergen Popp

Beilstein J. Nanotechnol. 2017, 8, 1183–1190, doi:10.3762/bjnano.8.120

• % humidity and 5% carbon dioxide in air. 75 cm2 cell culture flasks (658170; Greiner Bio-One GmbH, Germany) were used for cultivation of the cell lines. Every two or three days the medium was changed until approximately 100% confluence was reached. Cells were detached from the substrate by a 0.05% of trypsin

Uptake of the proteins HTRA1 and HTRA2 by cells mediated by calcium phosphate nanoparticles

• Olga Rotan,
• Katharina N. Severin,
• Simon Pöpsel,
• Alexander Peetsch,
• Melisa Merdanovic,
• Michael Ehrmann and
• Matthias Epple

Beilstein J. Nanotechnol. 2017, 8, 381–393, doi:10.3762/bjnano.8.40

• organisms [29]. HtrAs are unique serine proteases. Besides their trypsin-like protease domain, they possess at least one C-terminal PDZ domain and can form higher oligomers [30]. The main functions of its family members are key aspects of the protein quality control process [29]. The best-studied members of

• protein by HTRA2 induces autophagy, resulting in the clearance of damaged mitochondria. Similar as for HTRA1, the functional unit of HTRA2 is a trimer. Each protomer contains a trypsin-like protease domain and one C-terminal PDZ domain. The proteolytic activity can be modulated by binding of the PDZ

• with protein or the protein alone were added to the corresponding cell samples and incubated for another 3 h. The cells were then harvested for flow cytometry analysis as follows. All samples were washed three times with PBS, detached with trypsin/EDTA (3 min at 37 °C) and transferred into 15 mL Falcon

Facile fabrication of luminescent organic dots by thermolysis of citric acid in urea melt, and their use for cell staining and polyelectrolyte microcapsule labelling

• Nadezhda M. Zholobak,
• Anton L. Popov,
• Alexander B. Shcherbakov,
• Nelly R. Popova,
• Mykhailo M. Guzyk,
• Valeriy P. Antonovich,
• Alla V. Yegorova,
• Yuliya V. Scrypynets,
• Inna I. Leonenko,
• Alexander Ye. Baranchikov and
• Vladimir K. Ivanov

Beilstein J. Nanotechnol. 2016, 7, 1905–1917, doi:10.3762/bjnano.7.182

• , Russia). Cultures were incubated at 37 °C in air containing 5% CO2. Cells growing exponentially were harvested by a brief incubation with 0.25% trypsin–ethylenediaminetetraacetic acid (EDTA) solution (Gibco). The cellular uptake of microcapsules was studied using RAW 264.7 murine macrophage-like cell

Functional diversity of resilin in Arthropoda

• Jan Michels,
• Esther Appel and
• Stanislav N. Gorb

Beilstein J. Nanotechnol. 2016, 7, 1241–1259, doi:10.3762/bjnano.7.115

• completely dried, it loses its rubber-like characteristics and becomes relatively hard and brittle. Proteolytic enzymes such as pepsin or trypsin can be applied to test for the presence and distribution of resilin, because resilin is known to be digested by such enzymes. Resilin has been shown to be stained

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

• : RPMI-1640 medium; antibiotics (BioloT, Russia); trypsin, L-glutamine; PBS buffer (Sigma, USA); and fetal calf serum (Gibco, USA). TiO2 nanoparticles were synthesized in the crystal form (anatase) as described in [17]. Сhicken erythrocytes, MDCK cells, and influenza A virus strains Aichi/2/68 (H3N2), A

• /chicken/Kurgan/05/2005(H5N1), and А/Salekhard/01/2009 (H1N1) were from FBRI Vector, Russia. Trypsin (1 mg/mL) and penicillin-streptomycin (100 U/mL) (Sigma-Aldrich, USA) were stored at −80 °C. The IAV strains were grown in the allantoic cavity of 10-day-old embryonated chicken eggs at 37 °C. Allantoic

• -well plates (100 μL/well) and incubated at 37 °С, 5% CO2, and 100% humidity. The cells at ≈80% confluence were initially infected with one of the IAV subtypes, which was added into each well in RPMI-1640 medium (100 μL) containing trypsin (2 μg/mL) at a multiple infection of 0.1 TCID50/cell. The

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

• % trypsin overnight at 4 °C. The following day tissue was incubated for 1 h at 37 °C to separate epidermis from dermis. The epidermis was removed and the remaining dermis placed upside down in a 75 cm3 flask in RPMI 1640 medium (Invitrogen), then placed in a 37 °C incubator containing 5% CO2. After five

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

• ), pyruvic acid, silver nitrate, NaN3, sodium chloride, sodium dodecyl sulfate (SDS), sodium hydroxide (NaOH), sodium sarcosinate, streptomycin sulfate, sulforhodamine B (SRB), 1,1,3,3-tetramethoxypropane, MTT, thiobarbituric acid (TBA), trichloroacetic acid (TCA), Triton X-100, trizma-Base, trypsin/EDTA (5

• , 1 mM Na-pyruvate, 100 U/mL penicillin/streptomycin at 5% CO2 and 37 °C in humidified environment. Cells were harvested by trypsinization with 1 mL 0.5 mM trypsin/EDTA for 1 min at room temperature. Screening for photo-oxidative effect of NPs The nanoparticle stock suspensions (1 mg/mL) were prepared

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

• from American Tissue Culture Collection (ATCC). Dulbecco's Modified Eagle Medium (DMEM), fetal bovine serum (FBS), trypsin-EDTA (1×) and penicillin-streptomycin (100×) were purchased from PAA (Pasching, Austria). 2-(3,5-diphenyltetrazol-2-ium-2-yl)-4,5-dimethyl-1,3-thiazole bromide (MTT) was purchased

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

• medium without detaching cells from the culture surface. Afterward cells were detached with trypsin/EDTA, counted and cell pellets were resuspended in concentrated hydrochloric acid. Treatment with hydrochloric acid and ultrasound for 10 min destroyed the cells and dissociated the iron cores of SPIONs