MilliporeSigma
  • Genotoxicity of polyvinylpyrrolidone-coated silver nanoparticles in BEAS 2B cells.

Genotoxicity of polyvinylpyrrolidone-coated silver nanoparticles in BEAS 2B cells.

Toxicology (2012-11-13)
Penny Nymark, Julia Catalán, Satu Suhonen, Hilkka Järventaus, Renie Birkedal, Per Axel Clausen, Keld Alstrup Jensen, Minnamari Vippola, Kai Savolainen, Hannu Norppa
ABSTRACT

Silver nanoparticles (AgNPs) are widely utilized in various consumer products and medical devices, especially due to their antimicrobial properties. However, several studies have associated these particles with toxic effects, such as inflammation and oxidative stress in vivo and cytotoxic and genotoxic effects in vitro. Here, we assessed the genotoxic effects of AgNPs coated with polyvinylpyrrolidone (PVP) (average diameter 42.5±14.5 nm) on human bronchial epithelial BEAS 2B cells in vitro. AgNPs were dispersed in bronchial epithelial growth medium (BEGM) with 0.6 mg/ml bovine serum albumin (BSA). The AgNP were partially well-dispersed in the medium and only limited amounts (ca. 0.02 μg Ag(+) ion/l) could be dissolved after 24h. The zeta-potential of the AgNPs was found to be highly negative in pure water but was at least partially neutralized in BEGM with 0.6 mg BSA/ml. Cytotoxicity was measured by cell number count utilizing Trypan Blue exclusion and by an ATP-based luminescence cell viability assay. Genotoxicity was assessed by the alkaline single cell gel electrophoresis (comet) assay, the cytokinesis-block micronucleus (MN) assay, and the chromosomal aberration (CA) assay. The cells were exposed to various doses (0.5-48 μg/cm(2) corresponding to 2.5-240 μg/ml) of AgNPs for 4 and 24 h in the comet assay, for 48 h in the MN assay, and for 24 and 48 h in the CA assay. DNA damage measured by the percent of DNA in comet tail was induced in a dose-dependent manner after both the 4-h and the 24-h exposures to AgNPs, with a statistically significant increase starting at 16 μg/cm(2) (corresponding to 60.8 μg/ml) and doubling of the percentage of DNA in tail at 48 μg/cm(2). However, no induction of MN or CAs was observed at any of the doses or time points. The lack of induction of chromosome damage by the PVP-coated AgNPs is possibly due to the coating which may protect the cells from direct interaction with the AgNPs, either by reducing ion leaching from the particles or by causing extensive agglomeration of the nanoparticles, with a possible reduction of the cellular uptake.

MATERIALS
Product Number
Brand
Product Description

Supelco
Poly(vinylpolypyrrolidone), ~110 μm particle size
Sigma-Aldrich
Kollidon® 25
Povidone, European Pharmacopoeia (EP) Reference Standard
Supelco
Povidone, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Povidone, meets USP testing specifications
Sigma-Aldrich
1-Vinyl-2-pyrrolidinone, contains sodium hydroxide as inhibitor, ≥99%
Sigma-Aldrich
Polyvinylpyrrolidone, powder, average Mw ~29,000
Sigma-Aldrich
Polyvinylpyrrolidone, powder, average Mw ~55,000
Sigma-Aldrich
Polyvinylpyrrolidone, K 30
Sigma-Aldrich
Polyvinylpyrrolidone solution, K 60, 45% in H2O
Sigma-Aldrich
Polyvinylpyrrolidone, K 90
Sigma-Aldrich
Polyvinylpyrrolidone, average Mw ~1,300,000 by LS
Sigma-Aldrich
Polyvinylpyrrolidone, K 25, tested according to Ph. Eur.
Sigma-Aldrich
Polyvinylpyrrolidone, for molecular biology, nucleic acid hybridization tested, mol wt 360,000
Sigma-Aldrich
Polyvinylpyrrolidone, mol wt (number average molecular weight Mn 360kDa)
Sigma-Aldrich
Polyvinylpyrrolidone, average mol wt 40,000
Sigma-Aldrich
Polyvinylpyrrolidone, suitable for plant cell culture, average mol wt 10,000
Sigma-Aldrich
Polyvinylpyrrolidone, powder, BioXtra, suitable for mouse embryo cell culture
Sigma-Aldrich
Polyvinylpyrrolidone, average mol wt 10,000