title: In vitro screening of silver nanoparticles and ionic silver using neural networks yields differential effects on spontaneous activity and pharmacological responses.
abstract: Silver nanoparticles (AgNPs) are used in a wide range of consumer and medical products because of their antimicrobial and antifungal properties, and can translocate to the brain following exposure. Therefore, to screen AgNPs for potential impacts on human health, it is essential to examine neural function. The present study examined AgNPs (3 citrate coated, 3 PVP coated; 10-75nm) and AgNO3 effects on spontaneous and pharmacologically-induced neural network function in rat primary cortical cells on multi-well microelectrode array (mwMEA) plates. Baseline activity (1h) was recorded prior to exposure to non-cytotoxic concentrations of AgNPs and AgNO3 (0.08-0.63 and 0.08-1.7μg/ml, respectively). Changes in number of total extracellularly-recorded action potential spikes (total spikes; TS) and active electrodes (AE), relative to controls, were assessed 1, 24, and 48h after exposure to AgNP suspensions or AgNO3. After the 48h recording, the response to a pharmacological challenge with the GABAA antagonist, bicuculline (BIC), was assessed. Only two particles altered neural network function. Citrate coated 10nm AgNP caused concentration-related increases in AEs at 24h. After BIC treatment, PVP coated 75nm AgNP caused concentration-dependent increases in AE. AgNO3 effects differed from AgNPs, causing a concentration-related decrease in AEs at 24 and 48h, and a concentration-related decrease in TS following BIC challenge. Importantly, the direction of AgNO3 effects on neural activity was opposite those of 10nm Ag citrate at concentrations up to 0.63μg/ml, and different from 75nm Ag PVP, indicating ionic silver does not mediate these effects. These results demonstrate that non-cytotoxic concentrations of 10nm citrate- and 75nm PVP-coated Ag NPs alter neural network function in vitro, and should be considered for additional neurotoxicity hazard characterization.
authors.affiliation: Office of Research and Development, National Health and Environmental Effects Research Laboratory, US Environmental Protection Agency, Research Triangle Park, NC, United States, United States.
authors.forename: Jenna D
authors.initials: JD
authors.lastname: Strickland
authors.affiliation: Office of Research and Development, National Health and Environmental Effects Research Laboratory, US Environmental Protection Agency, Research Triangle Park, NC, United States, United States.
authors.forename: William R
authors.initials: WR
authors.lastname: LeFew
authors.affiliation: Office of Research and Development, National Health and Environmental Effects Research Laboratory, US Environmental Protection Agency, Research Triangle Park, NC, United States, United States.
authors.forename: James
authors.initials: J
authors.lastname: Crooks
authors.affiliation: Office of Research and Development, National Health and Environmental Effects Research Laboratory, US Environmental Protection Agency, Research Triangle Park, NC, United States, United States.
authors.forename: Diana
authors.initials: D
authors.lastname: Hall
authors.affiliation: Office of Research and Development, National Health and Environmental Effects Research Laboratory, US Environmental Protection Agency, Research Triangle Park, NC, United States, United States.
authors.forename: Jayna N R
authors.initials: JN
authors.lastname: Ortenzio
authors.affiliation: Office of Research and Development, National Health and Environmental Effects Research Laboratory, US Environmental Protection Agency, Research Triangle Park, NC, United States, United States.
authors.forename: Kevin
authors.initials: K
authors.lastname: Dreher
authors.affiliation: Office of Research and Development, National Health and Environmental Effects Research Laboratory, US Environmental Protection Agency, Research Triangle Park, NC, United States, United States. Electronic address: Shafer.tim@epa.gov.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Shafer
mesh_terms: D000200:Action Potentials / Q000187:drug effects*; D000818:Animals; D001640:Bicuculline / Q000494:pharmacology; D002951:Citrates / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D053768:Metal Nanoparticles / Q000008:administration & dosage / Q000633:toxicity*; D009415:Nerve Net / Q000187:drug effects*; D010316:Particle Size; D011205:Povidone / Q000737:chemistry; D051381:Rats; D012834:Silver / Q000008:administration & dosage / Q000633:toxicity*; D012835:Silver Nitrate / Q000008:administration & dosage / Q000633:toxicity*; D013997:Time Factors
chemical_list: D002951:Citrates; D012834:Silver; D012835:Silver Nitrate; D011205:Povidone; D001640:Bicuculline
keywords: Function; Microelectrode arrays; Nanoparticles; Neurophysiology; Screening; Silver