TY - JOUR
T1 - Pan-European inter-laboratory studies on a panel of in vitro cytotoxicity and pro-inflammation assays for nanoparticles
AU - Piret, Jean Pascal
AU - Bondarenko, Olesja M.
AU - Boyles, Matthew S P
AU - Himly, Martin
AU - Ribeiro, Ana R.
AU - Benetti, Federico
AU - Smal, Caroline
AU - Lima, Braulio
AU - Potthoff, Annegret
AU - Simion, Monica
AU - Dumortier, Elise
AU - Leite, Paulo Emilio C
AU - Balottin, Luciene Bottentuit
AU - Granjeiro, José Mauro
AU - Ivask, Angela
AU - Kahru, Anne
AU - Radauer-Preiml, Isabella
AU - Tischler, Ulrike
AU - Duschl, Albert
AU - Saout, Christelle
AU - Anguissola, Sergio
AU - Haase, Andrea
AU - Jacobs, An
AU - Nelissen, Inge
AU - Misra, Superb K.
AU - Toussaint, Olivier
PY - 2016/12/10
Y1 - 2016/12/10
N2 - The rapid development of nanotechnologies and increased production and use of nanomaterials raise concerns about their potential toxic effects for human health and environment. To evaluate the biological effects of nanomaterials, a set of reliable and reproducible methods and development of standard operating procedures (SOPs) is required. In the framework of the European FP7 NanoValid project, three different cell viability assays (MTS, ATP content, and caspase-3/7 activity) with different readouts (absorbance, luminescence and fluorescence) and two immune assays (ELISA of pro-inflammatory cytokines IL1-β and TNF-α) were evaluated by inter-laboratory comparison. The aim was to determine the suitability and reliability of these assays for nanosafety assessment. Studies on silver and copper oxide nanoparticles (NPs) were performed, and SOPs for particle handling, cell culture, and in vitro assays were established or adapted. These SOPs give precise descriptions of assay procedures, cell culture/seeding conditions, NPs/positive control preparation and dilutions, experimental well plate preparation, and evaluation of NPs interference. The following conclusions can be highlighted from the pan-European inter-laboratory studies: Testing of NPs interference with the toxicity assays should always be conducted. Interference tests should be designed as close as possible to the cell exposure conditions. ATP and MTS assays gave consistent toxicity results with low inter-laboratory variability using Ag and CuO NPs and different cell lines and therefore, could be recommended for further validation and standardization. High inter-laboratory variability was observed for Caspase 3/7 assay and ELISA for IL1-β and TNF-α measurements.
AB - The rapid development of nanotechnologies and increased production and use of nanomaterials raise concerns about their potential toxic effects for human health and environment. To evaluate the biological effects of nanomaterials, a set of reliable and reproducible methods and development of standard operating procedures (SOPs) is required. In the framework of the European FP7 NanoValid project, three different cell viability assays (MTS, ATP content, and caspase-3/7 activity) with different readouts (absorbance, luminescence and fluorescence) and two immune assays (ELISA of pro-inflammatory cytokines IL1-β and TNF-α) were evaluated by inter-laboratory comparison. The aim was to determine the suitability and reliability of these assays for nanosafety assessment. Studies on silver and copper oxide nanoparticles (NPs) were performed, and SOPs for particle handling, cell culture, and in vitro assays were established or adapted. These SOPs give precise descriptions of assay procedures, cell culture/seeding conditions, NPs/positive control preparation and dilutions, experimental well plate preparation, and evaluation of NPs interference. The following conclusions can be highlighted from the pan-European inter-laboratory studies: Testing of NPs interference with the toxicity assays should always be conducted. Interference tests should be designed as close as possible to the cell exposure conditions. ATP and MTS assays gave consistent toxicity results with low inter-laboratory variability using Ag and CuO NPs and different cell lines and therefore, could be recommended for further validation and standardization. High inter-laboratory variability was observed for Caspase 3/7 assay and ELISA for IL1-β and TNF-α measurements.
KW - In vitro assays
KW - Inter-laboratory studies
KW - Interference
KW - Nanoparticles
KW - Standard operating procedures
UR - http://www.scopus.com/inward/record.url?scp=85006355458&partnerID=8YFLogxK
U2 - 10.1007/s00204-016-1897-2
DO - 10.1007/s00204-016-1897-2
M3 - Article
AN - SCOPUS:85006355458
SN - 0340-5761
SP - 1
EP - 16
JO - Archives of Toxicology
JF - Archives of Toxicology
ER -