Cells were seeded on 96-well plates at a density of 3500–5000 cells per well (depending upon cell line) in 100 µL of complete media, recovered for 24 h, and then were exposed to AgNPs or AgNO3 (Sigma-Aldrich, St. Louis, MO, USA) in 100 µL of complete media containing doses of AgNPs or AgNO3 as listed in the figures. After 72 h, media containing AgNPs or AgNO3 were replaced with 200 µL of media containing 0.5 mg/mL 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT; Sigma-Aldrich) and incubated for 1 h at 37 °C. Medium was removed by vacuum suction, and cells were lysed in 200 µL of DMSO and absorbance read using a Molecular Devices Emax Precision Microplate Reader (San Jose, CA, USA) at 560 nm. Absorbance measurements at 650 nm were subtracted from each reading to correct for any inconsistencies in optical properties between wells.
Agnps
Manufactured by Merck Group
Sourced in United States, United Kingdom
AgNPs (Silver Nanoparticles) are a type of nanomaterial composed of silver particles with dimensions typically ranging from 1 to 100 nanometers. They exhibit unique physical, chemical, and antimicrobial properties due to their small size and high surface-to-volume ratio.
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Lab products found in correlation
Cu nps, by Merck Group (3 mentions)
Agno3, by Merck Group (3 mentions)
Zetasizer nano z, by Malvern Panalytical (2 mentions)
Zno np, by Merck Group (2 mentions)
Tio2 np, by Merck Group (2 mentions)
Attune nxt, by Thermo Fisher Scientific (2 mentions)
7700 agilent icp ms spectrometer, by Agilent Technologies (1 mentions)
Micromist nebulizer, by Agilent Technologies (1 mentions)
Silver nitrate agno3, by Sinopharm (1 mentions)
H 7650, by Hitachi (1 mentions)
D8 advance, by Bruker (1 mentions)
Ultrapure water, by Merck Group (1 mentions)
Agnps, by Malvern Panalytical (1 mentions)
Mapk family antibody sampler kit, by Cell Signaling Technology (1 mentions)
Phospho mapk family antibody sampler kit, by Cell Signaling Technology (1 mentions)
N acetylcysteine, by Merck Group (1 mentions)
Silver nitrate agno3, by Merck Group (1 mentions)
Sb203580, by Beyotime (1 mentions)
Atcc 25922, by American Type Culture Collection (1 mentions)
Staphylococcus epidermidis, by American Type Culture Collection (1 mentions)
37 protocols using agnps
1
MTT Assay for Assessing Silver Nanoparticle Cytotoxicity
2
Characterization of Monodisperse AgNPs
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AgNPs (size less than 40 nm) were obtained from Sigma-Aldrich (730793; St. Louis, MO, USA). According to the manufacturer instructions, AgNPs characterization was performed using transmission electron microscopy (TEM), dynamic light scattering (DLS), and UV/Visible spectral analysis to ensure that the monodisperse AgNPs are free from agglomeration with a density of 0.986 g/mL at 25 °C; refractive index n20/D 1.333; uorescence-λ em 401 nm) (https://www.sigmaaldrich.com/EG/en/product/aldrich/730793).
3
Nanoparticle Production by Pulsed Laser Ablation
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Nanoparticle production by pulsed picosecond laser ablation in deionised water (dH2O) was described in our previous publication [21 (link)]. Briefly, Ag plates (dimensions of 25 mm × 25 mm × 2 mm, purity 99.99%) were sterilised by immersion into ethanol and then autoclaved with dH2O. The Ag plates were then placed into a glass vessel containing 20 mL of dH2O at a level of 2 mm above them. A picosecond pulsed Nd: YVO4 laser with a wavelength of 1064 nm was used to ablate the plate at a pulse repetition rate of 200 kHz and an average power of 9.12 W. The average size for AgNPs produced was 27.2 nm, ranging from 10–70 nm [21 (link)].
The chemically produced AgNPs (concentration 20 μg/mL and average size about 35 nm) were purchased from Sigma-Aldrich (Dorset, UK).
The chemically produced AgNPs (concentration 20 μg/mL and average size about 35 nm) were purchased from Sigma-Aldrich (Dorset, UK).
4
Silver Nanoparticle Characterization
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5
Synthesis and Characterization of Silver Nanoparticles
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Unless stated otherwise, chemicals and solvents were purchased from Merck (Gillingham, UK). Anhydrous tetrahydrofuran (THF), 4,4′-methylene bis(phenyl isocyanate) (98%), 4-(2-aminoethyl)morpholine (99%) were used as received. AgNPs (Merck, Gillingham, UK) were less than 100 nm in diameter. KrasolTM HLBH-P2000 was supplied by Cray Valley, Saint- Avold, France.
6
ICP-MS Characterization of Silver Nanoparticles
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A 7700 Agilent ICP-MS spectrometer (Agilent Technologies, Tokyo, Japan) equipped with MicroMist™ nebulizer and Peltier cooled double-pass spray chamber was used for the characterization of silver nanoparticles at 107 amu. The ICP-MS operating parameters were as follows: RF power—1.55 kW; sample flow rate—0.336 mL min−1; carrier Ar gas flow rate—1.2 L min−1; acquisition time 60 s; dwell time 5 ms; and transport efficiency 0.032. The transport efficiency was determined by the particle-size method [66 (link)]. Ultrapure water (UPW) was used throughout the experiments (PURELAB Chorus 2+ (ELGA Veolia) water purification system). For the sonication of silver colloids, an ultrasonic bath (Kerry US) was used. Reference materials (RMs) of citrate-stabilized silver dispersions Ag NPs (Merck Bulgaria EAD) with mean size 40 ± 4 nm and total mass concentration of silver 0.02 mg mL−1 were used in this study for transport efficiency determination and calibration.
A standard solution of Ag 9.974 ± 0.041 mg L−1 in 2% HNO3 (CPAchem Ltd., Bogomilovo, Bulgaria) was used for the preparation of ionic standards.
A standard solution of Ag 9.974 ± 0.041 mg L−1 in 2% HNO3 (CPAchem Ltd., Bogomilovo, Bulgaria) was used for the preparation of ionic standards.
7
Drop-casting Ag Nanoparticles on Samples
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The aqueous solution of Ag NPs (Sigma-Aldrich Co.) with the concentrations of 0.02 mg/mL was drop-casted on the sample and then dried in the atmosphere for several minutes (i.e., LED III).
8
Synthesis and Characterization of Silver Nanoparticles
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Alkaline lignin was kindly supplied by Shan Dong Sun Paper Industry Joint Stock Co., Ltd. Silver nitrate (AgNO3) was obtained from Sinopharm Chemical Reagent CO., Ltd. (Shanghai, China). Ag NPs were purchased from Sigma-Aldrich, Co., Ltd. (< 100 nm particle size, 99.5% trace metals basis). Ammonium hydroxide (NH3·H2O, AR) and sodium hydroxide (NaOH, AR) were purchased from the Damao Chemical Reagent Factory (Tianjin, China).
9
Ultrastructural Alterations in HepG2 Cells Exposed to AgNPs
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Ultrastructural alterations in HepG2 cells exposed to AgNPs were visualized by TEM. For preparation of samples, HepG2 cells were seeded in 6-well plates at a density of 1 × 106 cells/well and incubated at 37°C and 5% CO2 for 24 hr. Cells were treated with 0.5, 2, or 6 μg/mL AgNPs (Sigma-Aldrich) and incubated for an additional 24 hr. Cells treated with silver nanoparticles were harvested by trypsinization and centrifugation. Samples of HepG2 cells were fixed in 2% glutaraldehyde for 2 hr at room temperature and then rinsed with phosphate buffer. Next, samples were postfixed in 2% osmium tetraoxide solution and subsequently dehydrated with ethanol. Samples were then embedded in epoxy resin and blocks were polymerized at 60°C overnight. After polymerization, thick sections of samples were selected for staining with toluidine blue and examined under a light microscope. Ultrathin sections of these samples were mounted on copper grids and counterstained with 2% uranyl acetate and lead citrate. Stained sections were examined using a Hitachi transmission electron microscope H-7650 at an operating voltage of 100 kV.
10
Characterization of Silver Nanoparticles
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Ag-NPs < 100 nm (99.5% pure) were purchased from Sigma (Steinheim, Germany). They have been previously characterized58 (link). Before use, each Ag-NP stock was mixed several times and an aliquot removed as a working solution that was sonicated 15 min in alternating cycles (2 × 30 s) ultrasonic sonicator. The physicochemical properties of Ag-NPs were confirmed by transmission electron microscopy (TEM) coupled with a microanalysis characterization (TECNAI G20, Ultra-Twin, FSB, Bizerte, Tunisia) and ultraviolet–visible (UV–Vis) spectroscopy (T60; PGInstruments, Leicestershire, UK). X-ray diffraction (XRD) characterization was performed using a D8 Advance diffractometer (Bruker, Bizerte), with analyses performed in Bragg–Brentano configuration at 40 kV and 40 mA.
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