title: Less is more: long-term in vitro exposure to low levels of silver nanoparticles provides new insights for nanomaterial evaluation.
abstract: In view of the vast number of new nanomaterials (NMs) that require testing and the constraints associated with animal models, the majority of studies to elucidate nanotoxicological effects have occurred in vitro, with limited correlation and applicability to in vivo systems and realistic, occupational exposure scenarios. In this study, we developed and implemented a chronic in vitro model coupled with lower, regulatory dosages in order to provide a more realistic assessment of NM-dependent consequences and illuminate the implications of long-term NM exposure. When keratinocytes were exposed to 50 nm silver nanoparticles (Ag-NPs), we determined that chronically dosed cells operated under augmented stress and modified functionality in comparison to their acute counterparts. Specifically, Ag-NP exposure through a chronic mechanism increased p38 activation, actin disorganization, heightened ki67 expression, and extensive gene modification. Additionally, chronic Ag-NP exposure altered the way in which cells perceived and responded to epidermal growth factor stimulation, indicating a transformation of cell functionality. Most importantly, this study demonstrated that chronic exposure in the pg/mL range to Ag-NPs did not induce a cytotoxic response, but instead activated sustained stress and signaling responses, suggesting that cells are able to cope with prolonged, low levels of Ag-NP exposure. In summary, we demonstrated that through implementation of a chronic dosimetry paradigm, which more closely resembles realistic NM exposure scenarios, it is possible to illuminate long-term cellular consequences, which greatly differ from previously obtained acute assessments.
authors.affiliation: Molecular Bioeffects Branch, Air Force Research Laboratory, Wright-Patterson AFB , 2729 R Street, Bldg 837, Dayton, Ohio 45433, United States.
authors.forename: Kristen K
authors.initials: KK
authors.lastname: Comfort
authors.forename: Laura K
authors.initials: LK
authors.lastname: Braydich-Stolle
authors.forename: Elizabeth I
authors.initials: EI
authors.lastname: Maurer
authors.forename: Saber M
authors.initials: SM
authors.lastname: Hussain
mesh_terms: D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D066255:EGF Family of Proteins / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D053768:Metal Nanoparticles; D018384:Oxidative Stress / Q000187:drug effects; D010316:Particle Size; D015398:Signal Transduction / Q000187:drug effects; D012834:Silver / Q000737:chemistry* / Q000633:toxicity*; D013997:Time Factors; D018675:Toxicity Tests
chemical_list: D066255:EGF Family of Proteins; D012834:Silver