Methods of in vitro toxicology
, B. Pool-Zobel
, V. Baker
, M. Balls
, B.J. Blaauboer
, A. Boobis
, S. Kevekordes
, J.-C. Lhuguenot
, R. Pieters
, J. Kleiner
University of Kaiserslautern, Department of Chemistry Food Chemistry & Environmental Toxicology, PO Box 3049,D-67653 Kaiserslautern, Germany
Friedrich Schiller University of Jena, Department of Nutritional Toxicology, Institute of Nutrition, Dornburger str. 25, D-07743 Jena, Germany
Unilever Research Colworth, SEAC Toxicology Unit, Sharnbrook MK42 0BH Bedfordshire, UK
European Centre for Validation of Alternative Methods, Joint Research Center, Institute for Health and Consumer Protection,T.P. 580, I-21020 Ispra (Varese), Italy
Institute for Risk Assessment Sciences (IRAS), Division of Toxicology, Utrecht University, PO Box 80.176,NL-3508 TD Utrecht, The Netherlands
Imperial College, Section on Clinical Pharmacology, Division of Medicine, Hammersmith Campus, Ducane Road, London W12 0NN, UK
Istituto Superiore di Sanita `, Laboratorio Tossicologia Comparata e Ecotossicologia, Viale Regina Elena, 299, I-00161 Rome, Italy
Su ¨ dzucker AG Mannheim/Ochsenfurt, ZAFES, Wormer Str. 11, D-67283 Obrigheim, Germany
ENSBANA—Universite´ de Bourgogne, De´ partement de Biochimie et Toxicologie Alimentaire, Campus Universitaire Montmuzard, 1,Esplanade Erasme, F-21000 Dijon, France
ILSI Europe, Avenue E. Mounier 83, Box 6, B-1200, Brussels, Belgium
In vitro methods are common and widely used for screening and ranking chemicals, and have also been taken into accountsporadically for risk assessment purposes in the case of food additives. However, the range of food-associated compounds amenableto in vitro toxicology is considered much broader, comprising not only natural ingredients, including those from food preparation,but also compounds formed endogenously after exposure, permissible/authorised chemicals including additives, residues, supple-ments, chemicals from processing and packaging and contaminants. A major promise of in vitro systems is to obtain mechanism-derived information that is considered pivotal for adequate risk assessment. This paper critically reviews the entire process of riskassessment by in vitro toxicology, encompassing ongoing and future developments, with major emphasis on cytotoxicity, cellularresponses, toxicokinetics, modelling, metabolism, cancer-related endpoints, developmental toxicity, prediction of allergenicity, andﬁnally, development and application of biomarkers. It describes in depth the use of in vitro methods in strategies for characterisingand predicting hazards to the human. Major weaknesses and strengths of these assay systems are addressed, together with some keyissues concerning major research priorities to improve hazard identiﬁcation and characterisation of food-associated chemicals.
2002 ILSI. Published by Elsevier Science Ltd. All rights reserved.
Hazard identiﬁcation; Risk assessment; Food chemicals; In vitro methods; Biomarkers; Parallelogram approach0278-6915/02/$ - see front matter
2002 ILSI. Published by Elsevier Science Ltd. All rights reserved.PII: S0278-6915(01)00118-1Food and Chemical Toxicology 40 (2002) 193–236www.elsevier.com/locate/foodchemtox
ADI, acceptable daily intake; Ah, aryl hydrocarbon; BiP, immunoglobulin binding protein; CHEST, chicken embryo toxicityscreening test; Comet test, single-cell microgel electrophoretic assay; DNA, deoxyribonucleic acid; ELISA, enzyme-linked immunosorbent assay;ER, endoplasmic reticulum; ES, embryonic stem cells; EST, expressed sequence trap; EU, European Union; FETAX, frog embryo teratogenesisassay xenopus; FISH, ﬂuorescence in situ hybridization; Grps, glucose-regulated proteins; GSH, glutathione; HESI, ILSI Human and Environ-mental Sciences Institute; Hsp, heat shock protein; HMWC, high molecular weight chemical; ICCVAM, US Interagency Committee for the Vali-dation of Alternative Methods; IEF, isoelectric focusing; IgE, immunoglobulin; LMWC, low molecular weight chemical; LOEL, lowest-observed-eﬀect level; MALDI, matrix assisted laser-desorption ionisation; MALDI-TOF, matrix assisted laser desorption-ionisation time of ﬂight; MM,micromass; MTD, maximum tolerated dose; MT, metallothioneine; NRU, neutral red uptake; OECD, Organisation for Economic Cooperation andDevelopment; PB-TK, physiologically-based toxicokinetic model; PCR, polymerase chain reaction; PCNA, proliferating cell nuclear antigen; PCs,partition coeﬃcients; PMs, prediction models; QSAR, quantitative structure–activity relationship; RAST, radioallergosorbent test; mRNA, mes-senger-ribonucleic acid; ROS, reactive oxygen species; RSM, restriction site mutation assay; SAR, structure–activity relationship; SDS–PAGE,sodium dodecyl sulphate–polyacrylamide gel electrophoresis; SOP, standard operating procedure; TCDD, 2,3,7,8-tetrachlorodibenzo-
, dose at which 50% of cells are aﬀected; TTC, threshold of toxicological concern; Vd, apparent volume of distribution.* Corresponding author. Tel.: +32-2-771-00-14; fax: +32-2-762-00-44.
email@example.com (J. Kleiner).