Pan-European inter-laboratory studies on a panel of in vitro cytotoxicity and pro-inflammation assays for nanoparticles

Jean Pascal Piret, Olesja M. Bondarenko, Matthew S P Boyles, Martin Himly, Ana R. Ribeiro, Federico Benetti, Caroline Smal, Braulio Lima, Annegret Potthoff, Monica Simion, Elise Dumortier, Paulo Emilio C Leite, Luciene Bottentuit Balottin, José Mauro Granjeiro, Angela Ivask, Anne Kahru, Isabella Radauer-Preiml, Ulrike Tischler, Albert Duschl, Christelle SaoutSergio Anguissola, Andrea Haase, An Jacobs, Inge Nelissen, Superb K. Misra, Olivier Toussaint

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Pages (from-to)1-16
Number of pages16
JournalArchives of Toxicology
DOIs
Publication statusAccepted/In press - 10 Dec 2016

Fingerprint

Cytotoxicity
Nanoparticles
Assays
Inflammation
Caspase 7
Nanostructures
Caspase 3
Cell Culture Techniques
Adenosine Triphosphate
Enzyme-Linked Immunosorbent Assay
Cell culture
Nanostructured materials
Nanotechnology
Poisons
Toxicity
Luminescence
Cells
Silver oxides
Copper
Cell Survival

Keywords

  • In vitro assays
  • Inter-laboratory studies
  • Interference
  • Nanoparticles
  • Standard operating procedures

Cite this

Piret, Jean Pascal ; Bondarenko, Olesja M. ; Boyles, Matthew S P ; Himly, Martin ; Ribeiro, Ana R. ; Benetti, Federico ; Smal, Caroline ; Lima, Braulio ; Potthoff, Annegret ; Simion, Monica ; Dumortier, Elise ; Leite, Paulo Emilio C ; Balottin, Luciene Bottentuit ; Granjeiro, José Mauro ; Ivask, Angela ; Kahru, Anne ; Radauer-Preiml, Isabella ; Tischler, Ulrike ; Duschl, Albert ; Saout, Christelle ; Anguissola, Sergio ; Haase, Andrea ; Jacobs, An ; Nelissen, Inge ; Misra, Superb K. ; Toussaint, Olivier. / Pan-European inter-laboratory studies on a panel of in vitro cytotoxicity and pro-inflammation assays for nanoparticles. In: Archives of Toxicology. 2016 ; pp. 1-16.
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Piret, JP, Bondarenko, OM, Boyles, MSP, Himly, M, Ribeiro, AR, Benetti, F, Smal, C, Lima, B, Potthoff, A, Simion, M, Dumortier, E, Leite, PEC, Balottin, LB, Granjeiro, JM, Ivask, A, Kahru, A, Radauer-Preiml, I, Tischler, U, Duschl, A, Saout, C, Anguissola, S, Haase, A, Jacobs, A, Nelissen, I, Misra, SK & Toussaint, O 2016, 'Pan-European inter-laboratory studies on a panel of in vitro cytotoxicity and pro-inflammation assays for nanoparticles', Archives of Toxicology, pp. 1-16. https://doi.org/10.1007/s00204-016-1897-2

Pan-European inter-laboratory studies on a panel of in vitro cytotoxicity and pro-inflammation assays for nanoparticles. / Piret, Jean Pascal; Bondarenko, Olesja M.; Boyles, Matthew S P; Himly, Martin; Ribeiro, Ana R.; Benetti, Federico; Smal, Caroline; Lima, Braulio; Potthoff, Annegret; Simion, Monica; Dumortier, Elise; Leite, Paulo Emilio C; Balottin, Luciene Bottentuit; Granjeiro, José Mauro; Ivask, Angela; Kahru, Anne; Radauer-Preiml, Isabella; Tischler, Ulrike; Duschl, Albert; Saout, Christelle; Anguissola, Sergio; Haase, Andrea; Jacobs, An; Nelissen, Inge; Misra, Superb K.; Toussaint, Olivier.

In: Archives of Toxicology, 10.12.2016, p. 1-16.

Research output: Contribution to journalArticle

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

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DO - 10.1007/s00204-016-1897-2

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