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    An in vitro testing strategy towards mimicking the inhalation of high aspect ratio nanoparticles

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    Author
    Endes, C; Schmid, O; Kinnear, C; Mueller, S; Camarero-Espinosa, S; Vanhecke, D; Foster, EJ; Petri-Fink, A; Rothen-Rutishauser, B; Weder, C; ...
    Date
    2014-09-23
    Source Title
    Particle and Fibre Toxicology
    Publisher
    BMC
    University of Melbourne Author/s
    Kinnear, Calum
    Affiliation
    School of Chemistry
    Metadata
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    Document Type
    Journal Article
    Citations
    Endes, C., Schmid, O., Kinnear, C., Mueller, S., Camarero-Espinosa, S., Vanhecke, D., Foster, E. J., Petri-Fink, A., Rothen-Rutishauser, B., Weder, C. & Clift, M. J. D. (2014). An in vitro testing strategy towards mimicking the inhalation of high aspect ratio nanoparticles. PARTICLE AND FIBRE TOXICOLOGY, 11 (1), https://doi.org/10.1186/s12989-014-0040-x.
    Access Status
    Open Access
    URI
    http://hdl.handle.net/11343/254727
    DOI
    10.1186/s12989-014-0040-x
    Abstract
    BACKGROUND: The challenge remains to reliably mimic human exposure to high aspect ratio nanoparticles (HARN) via inhalation. Sophisticated, multi-cellular in vitro models are a particular advantageous solution to this issue, especially when considering the need to provide realistic and efficient alternatives to invasive animal experimentation for HARN hazard assessment. By incorporating a systematic test-bed of material characterisation techniques, a specific air-liquid cell exposure system with real-time monitoring of the cell-delivered HARN dose in addition to key biochemical endpoints, here we demonstrate a successful approach towards investigation of the hazard of HARN aerosols in vitro. METHODS: Cellulose nanocrystals (CNCs) derived from cotton and tunicates, with differing aspect ratios (~9 and ~80), were employed as model HARN samples. Specifically, well-dispersed and characterised CNC suspensions were aerosolised using an "Air Liquid Interface Cell Exposure System" (ALICE) at realistic, cell-delivered concentrations ranging from 0.14 to 1.57 μg/cm2. The biological impact (cytotoxicity, oxidative stress levels and pro-inflammatory effects) of each HARN sample was then assessed using a 3D multi-cellular in vitro model of the human epithelial airway barrier at the air liquid interface (ALI) 24 hours post-exposure. Additionally, the testing strategy was validated using both crystalline quartz (DQ12) as a positive particulate control in the ALICE system and long fibre amosite asbestos (LFA) to confirm the susceptibility of the in vitro model to a fibrous insult. RESULTS: A rapid (≤ 4 min), controlled nebulisation of CNC suspensions enabled a dose-controlled and spatially homogeneous CNC deposition onto cells cultured under ALI conditions. Real-time monitoring of the cell-delivered CNC dose with a quartz crystal microbalance was accomplished. Independent of CNC aspect ratio, no significant cytotoxicity (p>0.05), induction of oxidative stress, or (pro)-inflammatory responses were observed up to the highest concentration of 1.57 μg/cm2. Both DQ12 and LFA elicited a significant (p<0.05) pro-inflammatory response at sub-lethal concentrations in vitro. CONCLUSION: In summary, whilst the present study highlights the benign nature of CNCs, it is the advanced technological and mechanistic approach presented that allows for a state of the art testing strategy to realistically and efficiently determine the in vitro hazard concerning inhalation exposure of HARN.

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