TY - JOUR
T1 - Improving quality in nanoparticle-induced cytotoxicity testing by a tiered inter-laboratory comparison study
AU - Nelissen, Inge
AU - Haase, Andrea
AU - Anguissola, Sergio
AU - Rocks, Louise
AU - Jacobs, An
AU - Willems, Hanny
AU - Riebeling, Christian
AU - Luch, Andreas
AU - Piret, Jean Pascal
AU - Toussaint, Olivier
AU - Trouiller, Bénédicte
AU - Lacroix, Ghislaine
AU - Gutleb, Arno C.
AU - Contal, Servane
AU - Diabaté, Silvia
AU - Weiss, Carsten
AU - Lozano-Fernández, Tamara
AU - González-Fernández, África
AU - Dusinska, Maria
AU - Huk, Anna
AU - Stone, Vicki
AU - Kanase, Nilesh
AU - Nocuń, Marek
AU - Stępnik, Maciej
AU - Meschini, Stefania
AU - Ammendolia, Maria Grazia
AU - Lewinski, Nastassja
AU - Riediker, Michael
AU - Venturini, Marco
AU - Benetti, Federico
AU - Topinka, Jan
AU - Brzicova, Tana
AU - Milani, Silvia
AU - Rädler, Joachim
AU - Salvati, Anna
AU - Dawson, Kenneth A.
N1 - Funding Information:
Funding: This research received financial support from the EC FP7 project QualityNano (INFRA-2010-1.1.31-262163). The contribution of LIST was supported by the Fonds National de la Recherche of Luxembourg within the project NANION (FNR/12/SR/4009651). J.-P. Piret and O. Toussaint acknowledge the DG06 (Direction générale opérationnelle de l’Economie, de l’Emploi et de la Recherche) of the Walloon Region of Belgium for the project complement ‘QNano’ (2011-14) n°1117448. O. Toussaint, who unfortunately deceased in 2016, was a senior Research Associate from the Belgian F.R.S.-FNRS. The contribution of NILU was also supported by the NorNANoREG project (Norwegian Research Council, 239199/070). All authors are grateful to their institutions for supporting this work.
Funding Information:
This research received financial support from the EC FP7 project QualityNano (INFRA-2010-1.1.31-262163). The contribution of LIST was supported by the Fonds National de la Recherche of Luxembourg within the project NANION (FNR/12/SR/4009651). J.-P. Piret and O. Toussaint acknowledge the DG06 (Direction g?n?rale op?rationnelle de l?Economie, de l?Emploi et de la Recherche) of the Walloon Region of Belgium for the project complement ?QNano? (2011-14) n?1117448. O. Toussaint, who unfortunately deceased in 2016, was a senior Research Associate from the Belgian F.R.S.-FNRS. The contribution of NILU was also supported by the NorNANoREG project (Norwegian Research Council, 239199/070). All authors are grateful to their institutions for supporting this work.
Publisher Copyright:
© 2020 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2020/8
Y1 - 2020/8
N2 - The quality and relevance of nanosafety studies constitute major challenges to ensure their key role as a supporting tool in sustainable innovation, and subsequent competitive economic advantage. However, the number of apparently contradictory and inconclusive research results has increased in the past few years, indicating the need to introduce harmonized protocols and good practices in the nanosafety research community. Therefore, we aimed to evaluate if best-practice training and inter-laboratory comparison (ILC) of performance of the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay for the cytotoxicity assessment of nanomaterials among 15 European laboratories can improve quality in nanosafety testing. We used two well-described model nanoparticles, 40-nm carboxylated polystyrene (PS-COOH) and 50-nm amino-modified polystyrene (PS-NH2). We followed a tiered approach using well-developed standard operating procedures (SOPs) and sharing the same cells, serum and nanoparticles. We started with determination of the cell growth rate (tier 1), followed by a method transfer phase, in which all laboratories performed the first ILC on the MTS assay (tier 2). Based on the outcome of tier 2 and a survey of laboratory practices, specific training was organized, and the MTS assay SOP was refined. This led to largely improved intra-and inter-laboratory reproducibility in tier 3. In addition, we confirmed that PS-COOH and PS-NH2 are suitable negative and positive control nanoparticles, respectively, to evaluate impact of nanomaterials on cell viability using the MTS assay. Overall, we have demonstrated that the tiered process followed here, with the use of SOPs and representative control nanomaterials, is necessary and makes it possible to achieve good inter-laboratory reproducibility, and therefore high-quality nanotoxicological data.
AB - The quality and relevance of nanosafety studies constitute major challenges to ensure their key role as a supporting tool in sustainable innovation, and subsequent competitive economic advantage. However, the number of apparently contradictory and inconclusive research results has increased in the past few years, indicating the need to introduce harmonized protocols and good practices in the nanosafety research community. Therefore, we aimed to evaluate if best-practice training and inter-laboratory comparison (ILC) of performance of the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay for the cytotoxicity assessment of nanomaterials among 15 European laboratories can improve quality in nanosafety testing. We used two well-described model nanoparticles, 40-nm carboxylated polystyrene (PS-COOH) and 50-nm amino-modified polystyrene (PS-NH2). We followed a tiered approach using well-developed standard operating procedures (SOPs) and sharing the same cells, serum and nanoparticles. We started with determination of the cell growth rate (tier 1), followed by a method transfer phase, in which all laboratories performed the first ILC on the MTS assay (tier 2). Based on the outcome of tier 2 and a survey of laboratory practices, specific training was organized, and the MTS assay SOP was refined. This led to largely improved intra-and inter-laboratory reproducibility in tier 3. In addition, we confirmed that PS-COOH and PS-NH2 are suitable negative and positive control nanoparticles, respectively, to evaluate impact of nanomaterials on cell viability using the MTS assay. Overall, we have demonstrated that the tiered process followed here, with the use of SOPs and representative control nanomaterials, is necessary and makes it possible to achieve good inter-laboratory reproducibility, and therefore high-quality nanotoxicological data.
KW - Best practice
KW - Cytotoxicity
KW - Inter-laboratory comparison
KW - nanosafety
KW - Training
UR - http://www.scopus.com/inward/record.url?scp=85088306454&partnerID=8YFLogxK
U2 - 10.3390/nano10081430
DO - 10.3390/nano10081430
M3 - Article
AN - SCOPUS:85088306454
SN - 2079-4991
VL - 10
SP - 1
EP - 20
JO - Nanomaterials
JF - Nanomaterials
IS - 8
M1 - 1430
ER -