Laetitia Pele1, Carolin T Haas1, Rachel Hewitt1, Nuno Faria1, Andy Brown2, Jonathan Powell1. 1. 1Medical Research Council - Human Nutrition Research, Elsie Widdowson Laboratory, Fulbourn Road, Cambridge, CB1 9NL, UK. 2. 2Institute for Materials Research, SPEME, University of Leeds, Leeds, LS2 9JT, UK.
Abstract
AIM: To determine whether in vitro experimental conditions dictate cellular activation of the inflammasome by apatitic calcium phosphate nanoparticles. MATERIAL & METHODS: The responses of blood-derived primary human cells to in situ-formed apatite were investigated under different experimental conditions to assess the effect of aseptic culture, cell rest and duration of particle exposure. Cell death and particle uptake were assessed, while IL-1β and caspase 1 responses, with and without lipopolysaccharide prestimulation, were evaluated as markers of inflammasome activation. RESULTS: Under carefully addressed experimental conditions, apatitic nanoparticles did not induce cell death or engage the inflammasome platform, although both could be triggered through artefacts of experimentation. CONCLUSION: In vitro studies often predict that engineered nanoparticles, such as synthetic apatite, are candidates for inflammasome activation and, hence, are toxic. However, the experimental setting must be very carefully considered as it may promote false-positive outcomes.
AIM: To determine whether in vitro experimental conditions dictate cellular activation of the inflammasome by apatitic calcium phosphate nanoparticles. MATERIAL & METHODS: The responses of blood-derived primary human cells to in situ-formed apatite were investigated under different experimental conditions to assess the effect of aseptic culture, cell rest and duration of particle exposure. Cell death and particle uptake were assessed, while IL-1β and caspase 1 responses, with and without lipopolysaccharide prestimulation, were evaluated as markers of inflammasome activation. RESULTS: Under carefully addressed experimental conditions, apatitic nanoparticles did not induce cell death or engage the inflammasome platform, although both could be triggered through artefacts of experimentation. CONCLUSION: In vitro studies often predict that engineered nanoparticles, such as synthetic apatite, are candidates for inflammasome activation and, hence, are toxic. However, the experimental setting must be very carefully considered as it may promote false-positive outcomes.
Authors: Laetitia C Pele; Carolin T Haas; Rachel E Hewitt; Jack Robertson; Jeremy Skepper; Andy Brown; Juan Carlos Hernandez-Garrido; Paul A Midgley; Nuno Faria; Helen Chappell; Jonathan J Powell Journal: Nanomedicine Date: 2016-07-29 Impact factor: 5.307
Authors: Sina Köppert; Andrea Büscher; Anne Babler; Ahmed Ghallab; Eva M Buhl; Eicke Latz; Jan G Hengstler; Edward R Smith; Willi Jahnen-Dechent Journal: Front Immunol Date: 2018-09-04 Impact factor: 7.561
Authors: Sebastian Riedle; Laetitia C Pele; Don E Otter; Rachel E Hewitt; Harjinder Singh; Nicole C Roy; Jonathan J Powell Journal: Part Fibre Toxicol Date: 2017-12-08 Impact factor: 9.400