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

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

• grown to ≈80% confluency were detached from culture flasks by trypsin/EDTA and washed off with measurement media (cell line-specific media supplemented with 20 mM HEPES) containing 10% FCS. Cells were pelleted (420 g for 90 s) and resuspended in measurement media. Petri dishes with PDMS masks were

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

• agent ethylenediaminetetraacetic acid (EDTA, 2 mL) for 10 min in the incubator. Then, EDTA was removed and cells were detached from the substrate by incubation with trypsin/EDTA (1 mL) for 10 min in the incubator. Trypsination was stopped by addition of medium (10 mL), which was removed afterward by

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

• mixture of a variety of substances, it contains, among others, digestive enzymes like trypsin or chymotrypsin and also considerable amounts of immunoglobulin A and mucin [30]. It is known that components of the intestinal fluid, e.g., digestive enzymes from the gut lumen, can be immobilized at the outer

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

• Technologies) and detached from the culture flasks 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 hMSCs were collected and washed twice with RPMI1640/10% FCS. Determination of cellular Ag-NP

Imaging the intracellular degradation of biodegradable polymer nanoparticles

• Anne-Kathrin Barthel,
• Martin Dass,
• Melanie Dröge,
• Jens-Michael Cramer,
• Daniela Baumann,
• Markus Urban,
• Katharina Landfester,
• Volker Mailänder and
• Ingo Lieberwirth

Beilstein J. Nanotechnol. 2014, 5, 1905–1917, doi:10.3762/bjnano.5.201

• of ciprofloxacin (Fluka, Switzerland; 2 mg·mL−1, 0.6%). Cells were grown in 500 cm2 triple flasks (Nunc, Germany) in a humidified incubator at 37 °C and 5% CO2. The culture medium was changed twice a week. At confluence, cells were detached by 0.5% trypsin (Invitrogen, Germany) and seeded in the

• specified concentrations. Flow cytometry Flow cytometry was used for quantification of intracellular nanoparticles and for the analysis of cell viability. Similar to the procedures previously described [26], adherent cells were detached by trypsin (Gibco, Germany) and seeded in α-MEM at a density of 100 000

• , the cells were detached by trypsin and newly seeded out. At the specified residence times, the sapphire disks were removed from the wells and dipped into 1-hexadecane to remove the remaining medium. The disks were covered with aluminum disks to prevent squeezing of the cells and the sandwich was

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

• ) supplemented with SupplementMix (PromoCell GmbH, Germany). Both cell lines were cultured at 37 ºC in a 5% CO2 humidified environment and the growth medium was exchanged every 2–3 days. Once the cells reached 70–85% confluency they were subcultivated. To detach the cells, GIBCO® trypsin (Life Technologies GmbH

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

• ; Cfh: complement component factor h; Mug1: murinoglobulin 1; Pzp: pregnancy zone protein; Itih4: inter alpha-trypsin inhibitor, heavy chain 4; Gsn: gelsolin; Ahsg: alpha-2-HS-glycoprotein; Fn1: fibronectin 1; Alb: albumin; C3: complement component 3. Schematics of the analysis of quantitative NP

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

• , Switzerland) and 1% penicillin/streptomycin (Gibco, Luzern, Switzerland) and kept at 37 °C and 5% CO2. The A549 epithelial cells were split twice per week using trypsin (0.05% trypsin-EDTA, GIBCO, Switzerland). J774A.1 cells were sub-cultured using the scraping method, resuspended in the medium and finally

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

• Analytics (Z-PS-SIL-GFP-0.07, Landsberg am Lech, Germany). Dulbecco’s Modified Eagle Medium (DMEM), Roswell Park Memorial Institute medium (RPMI-1640), Hank’s Balanced Salt Solution (HBSS), Dulbecco’s Phosphate Buffered Saline without Ca2+ and Mg2+ (DPBS-/-), penicillin, streptomycin, and trypsin were from

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

• observed after the reaction for all of the proteins except trypsin (TRY) and glucose oxidase (GOX). The AuNPs formed in the presence of the different proteins were characterized by UV–vis spectroscopy (Figure 1). The addition of Ag ions played an important role in the formation of the AuNPs, which is

• histone (HIS), chicken egg white lysozyme (LYS), ovalbumin (OVA), bovine serum albumin (BSA), bovine hemoglobin (BHG), bovine gamma globulin (BGG), glucose oxidase (Aspergillus niger) (GOX), and trypsin (porcine pancreas, TRY), used for the experiments were obtained from Sigma (USA) as a lyophilized

Near-infrared dye loaded polymeric nanoparticles for cancer imaging and therapy and cellular response after laser-induced heating

• Tingjun Lei,
• Alicia Fernandez-Fernandez,
• Romila Manchanda,
• Yen-Chih Huang and
• Anthony J. McGoron

Beilstein J. Nanotechnol. 2014, 5, 313–322, doi:10.3762/bjnano.5.35

• -Aldrich (St. Louis, MI): Malic acid, 1,12-dodecanedioic acid (DDA), dimethylsulfoxide (DMSO > 99.9%, reagent grade), pluronic F-127, Dulbecco phosphate-buffered saline (DPBS), phosphate buffered saline (PBS), IR820, penicillin-streptomycin solution, tetrahydrofuran (THF) and trypsin-EDTA. Glycerol was

• with PBS and collected by incubating with trypsin for 5 min. The same number of cells were counted and incubated with CM-H2DCFDA in the dark. After 30 min, cells were briefly washed with PBS, and the intensity of DCF was measured by a flow cytometer (BD Accuri C6, NJ). Study of HIF-1 expression To

Magnetic nanoparticles for biomedical NMR-based diagnostics

• Huilin Shao,
• Tae-Jong Yoon,
• Monty Liong,
• Ralph Weissleder and
• Hakho Lee

Beilstein J. Nanotechnol. 2010, 1, 142–154, doi:10.3762/bjnano.1.17

• within the DEVD site, which led to a corresponding increase in T2 relaxation time (Figure 5c). This dissociation was not observed when a specific caspase-3 inhibitor was added. A similar reverse switching strategy has been used to detect trypsin, renin, and matrix metalloproteinase 2 activities [57