PMID- 26778741 OWN - NLM STAT- MEDLINE DCOM- 20161213 LR - 20161230 IS - 1879-3177 (Electronic) IS - 0887-2333 (Linking) VI - 32 DP - 2016 Apr TI - Prevalidation of the ex-vivo model PCLS for prediction of respiratory toxicity. PG - 347-61 LID - S0887-2333(16)30006-6 [pii] LID - 10.1016/j.tiv.2016.01.006 [doi] AB - In acute inhalation toxicity studies, animals inhale substances at given concentrations. Without additional information, however, appropriate starting concentrations for in-vivo inhalation studies are difficult to estimate. The goal of this project was the prevalidation of precision-cut lung slices (PCLS) as an ex-vivo alternative to reduce the number of animals used in inhalation toxicity studies. According to internationally agreed principles for Prevalidation Studies, the project was conducted in three independent laboratories. The German BfR provided consultancy in validation principles and independent support with biostatistics. In all laboratories, rat PCLS were prepared and exposed to 5 concentrations of 20 industrial chemicals under submerged culture conditions for 1h. After 23 h post-incubation, toxicity was assessed by measurement of released lactate dehydrogenase and mitochondrial activity. In addition, protein content and pro-inflammatory cytokine IL-1alpha were measured. For all endpoints IC50 values were calculated if feasible. For each endpoint test acceptance criteria were established. This report provides the final results for all 20 chemicals. More than 900 concentration-response curves were analyzed. Log10[IC50 (muM)], obtained for all assay endpoints, showed best intra- and inter-laboratory consistency for the data obtained by WST-1 and BCA assays. While WST-1 and LDH indicated toxic effects for the majority of substances, only some of the substances induced an increase in extracellular IL-1alpha. Two prediction models (two-group classification model, prediction of LC50 by IC50) were developed and showed promising results. CI - Copyright (c) 2016 The Authors. Published by Elsevier Ltd.. All rights reserved. FAU - Hess, A AU - Hess A AD - BASF SE, Experimental Toxicology and Ecology, 67056 Ludwigshafen, Germany. FAU - Wang-Lauenstein, L AU - Wang-Lauenstein L AD - Department of Pre-clinical Pharmacology and In Vitro Toxicology, Fraunhofer Institute for Toxicology and Experimental Medicine (ITEM), Nikolai-Fuchs-Str. 1, 30625 Hannover, Germany; German Center for Lung Research, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Germany. FAU - Braun, A AU - Braun A AD - Department of Pre-clinical Pharmacology and In Vitro Toxicology, Fraunhofer Institute for Toxicology and Experimental Medicine (ITEM), Nikolai-Fuchs-Str. 1, 30625 Hannover, Germany; German Center for Lung Research, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Germany. FAU - Kolle, S N AU - Kolle SN AD - BASF SE, Experimental Toxicology and Ecology, 67056 Ludwigshafen, Germany. FAU - Landsiedel, R AU - Landsiedel R AD - BASF SE, Experimental Toxicology and Ecology, 67056 Ludwigshafen, Germany. FAU - Liebsch, M AU - Liebsch M AD - German Federal Institute for Risk Assessment (BfR), Max-Dohrn-Str. 8-10, 10589 Berlin, Germany. FAU - Ma-Hock, L AU - Ma-Hock L AD - BASF SE, Experimental Toxicology and Ecology, 67056 Ludwigshafen, Germany. FAU - Pirow, R AU - Pirow R AD - German Federal Institute for Risk Assessment (BfR), Max-Dohrn-Str. 8-10, 10589 Berlin, Germany. FAU - Schneider, X AU - Schneider X AD - Institute of Pharmacology and Toxicology, Medical Faculty of RWTH Aachen University, Wendlingweg 2, Aachen, Germany. FAU - Steinfath, M AU - Steinfath M AD - German Federal Institute for Risk Assessment (BfR), Max-Dohrn-Str. 8-10, 10589 Berlin, Germany. FAU - Vogel, S AU - Vogel S AD - BASF SE, Experimental Toxicology and Ecology, 67056 Ludwigshafen, Germany. FAU - Martin, C AU - Martin C AD - Institute of Pharmacology and Toxicology, Medical Faculty of RWTH Aachen University, Wendlingweg 2, Aachen, Germany. Electronic address: chmartin@ukaachen.de. FAU - Sewald, K AU - Sewald K AD - Department of Pre-clinical Pharmacology and In Vitro Toxicology, Fraunhofer Institute for Toxicology and Experimental Medicine (ITEM), Nikolai-Fuchs-Str. 1, 30625 Hannover, Germany; German Center for Lung Research, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Germany. Electronic address: katherina.sewald@item.fraunhofer.de. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20160109 PL - England TA - Toxicol In Vitro JT - Toxicology in vitro : an international journal published in association with BIBRA JID - 8712158 RN - 0 (2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium) RN - 0 (Interleukin-1alpha) RN - 0 (Tetrazolium Salts) RN - EC 1.1.1.27 (L-Lactate Dehydrogenase) SB - IM MH - Animal Testing Alternatives MH - Animals MH - Cell Survival MH - Female MH - In Vitro Techniques MH - Interleukin-1alpha/metabolism MH - L-Lactate Dehydrogenase/metabolism MH - Laboratories MH - *Lung/metabolism MH - *Models, Biological MH - Rats, Wistar MH - Reproducibility of Results MH - Tetrazolium Salts/metabolism MH - *Toxicity Tests OTO - NOTNLM OT - Acute inhalation toxicity OT - Industrial chemicals OT - Precision-cut lung slices OT - Prevalidation OT - Reproducibility OT - Respiratory toxicity OT - Submersed culture conditions OT - Transferability OT - Two-group classification model EDAT- 2016/01/19 06:00 MHDA- 2016/12/15 06:00 CRDT- 2016/01/19 06:00 PHST- 2015/07/01 00:00 [received] PHST- 2016/01/05 00:00 [revised] PHST- 2016/01/07 00:00 [accepted] PHST- 2016/01/19 06:00 [entrez] PHST- 2016/01/19 06:00 [pubmed] PHST- 2016/12/15 06:00 [medline] AID - S0887-2333(16)30006-6 [pii] AID - 10.1016/j.tiv.2016.01.006 [doi] PST - ppublish SO - Toxicol In Vitro. 2016 Apr;32:347-61. doi: 10.1016/j.tiv.2016.01.006. Epub 2016 Jan 9.