Integrated Decision-tree Testing Strategies for Acute Systemic Toxicity and Toxicokinetics with Respect to the Requirements of the EU REACH Legislation
Restricted accessOtherFirst published online February, 2008
Integrated Decision-tree Testing Strategies for Acute Systemic Toxicity and Toxicokinetics with Respect to the Requirements of the EU REACH Legislation
Liverpool John Moores University and FRAME conducted a joint research project, sponsored by Defra, on the status of alternatives to animal testing with regard to the European Union REACH (Registration, Evaluation and Authorisation of Chemicals) system for the safety testing and risk assessment of chemicals. The project covered all the main toxicity endpoints associated with REACH. This paper focuses on the use of alternative (non-animal) methods (both in vitro and in silico) for acute systemic toxicity and toxicokinetic testing. The paper reviews in vitro tests based on basal cytotoxicity and target organ toxicity, along with QSAR models and expert systems available for this endpoint. The use of PBPK modelling for the prediction of ADME properties is also discussed. These tests are then incorporated into a decision-tree style, integrated testing strategy, which also includes the use of refined in vivo acute toxicity tests, as a last resort. The implementation of the strategy is intended to minimise the use of animals in the testing of acute systemic toxicity and toxicokinetics, whilst satisfying the scientific and logistical demands of the EU REACH legislation.
GrindonC., CombesR., CroninM.T.D., RobertsD.W. & GarrodJ. (2006). A review of the status of alternative approaches to animal testing and the development of integrated testing strategies for assessing the toxicity of chemicals under REACH —A summary of a DEFRA-funded project conducted by Liverpool John Moores University and FRAME. ATLA34Suppl. 1, 149–158.
2.
GrindonC., CombesR., CroninM.T.D., RobertsD.W. & GarrodJ.F. (2006). Integrated testing strategies for use in the EU REACH system. ATLA34, 407–427.
3.
BartonH.A., PastoorT.P., BaetckeK., ChambersJ.E., DilibertoJ., DoerrerN.G., DriverJ.H., HastingsC.E., IyengarS., KriegerR., StahlB. & TimchalkC. (2006). The acquisition and application of absorption, distribution, metabolism and excretion (ADME) data in agricultural chemical safety assessments. Critical Reviews in Toxicology36, 9–35.
4.
Anon. (2005). Globally Harmonised System of Classification and Labelling of Chemicals (GHS), 1st revised edition, Annex 2 — Classification and Labelling Summary Tables, 40pp. Geneva, Switzerland: United Nations Economic Commission for Europe (UNECE). Available at: http://www.unece.org/trans/danger/publi/ghs/ghs_rev01/01files_e.html (Accessed 27.09.06).
5.
Anon. (2006). Briefing Note on Number of Animals Expected to be Used Under REACH. Summary of Re-Assessment Performed by the JRC, 4pp. Ispra, Italy: European Commission Joint Research Centre. Available at: http://ihcp.jrc.it/docs/tcs/REACHanimalfigures.pdf (Accessed 04.10.07).
CombesR.D., GauntI. & BallsM. (2004). A scientific and animal welfare assessment of the OECD Health Effects Test Guidelines for the safety testing of chemicals under the European Union REACH system. ATLA32, 163–208.
13.
CombesR.D. (2006). The OECD Health Effects Test Guidelines for REACH need updating. ALTEX23 Special Issue, 124–130.
14.
ClemedsonC., McFarlane-AbdullaE., AnderssonM., BarileF.A., CallejaM.C., ChesnéC., ClothierR., CottinM., CurrenR., Daniel-SzolgayE., DierickxP., FerroM., FiskesjöG., Garza-OcanasL., Gómez-LechónM.J., GüldenM., IsomaaB., JanusJ., JudgeP., KahruA., KempR.B., KerszmanG., KristenU., KunimotoM., KärenlampiS., LavrijsenK., LewanL., LiliusH., OhnoT., PersooneG., RoguetR., RomertL., SawyerT., SeibertH., ShrivastavaR., StammatiA., TanakaN., Torres AlanisO., VossJ-U., WakuriS., WalumE., WangX., ZuccoF. & EkwallB. (1996). MEIC evaluation of acute systemic toxicity: Part I. Methodology of 68 in vitro toxicity assays used to test the first 30 reference chemicals. ATLA24, 251–272.
15.
EkwallB., ClemedsonC., CrafoordB., EkwallB., HallanderS., WalumE. & BondessonI. (1998). MEIC evaluation of acute systemic toxicity: Part V. Rodent and human toxicity data for the 50 reference chemicals. ATLA26, 571–616.
16.
ClemedsonC., BarileF.A., ChesnéC., CottinC., CurrenR., EkwallB., FerroM., Gómez-LechónM., ImaiK., JanusJ., KempR.B., KerszmanG., KjellstrandP., LavrijsenK., LogemannP., McFarlane-AbdullaE., RoguetR., SegnerH., ThuvanderA., WalumE. & EkwallB. (2000). MEIC evaluation of acute systemic toxicity: Part VII. Prediction of human toxicity by results from testing of the first 30 reference chemicals with 27 further in vitro assays. ATLA28, 161–200.
17.
EkwallB. (1999). Overview of the final MEIC results. II: The in vitro–in vivo evaluation, including the selection of a practical battery of cell tests for prediction of acute lethal blood concentrations in humans. Toxicology in Vitro13, 665–673.
18.
Anon. (2001). Report of the International Workshop on In Vitro Methods for Assessing Acute Systemic Toxicity, NIH Publication No. 01-4499, 186pp. Research Triangle Park, NC, USA:NIEHS.
19.
SpielmannH., GenschowE., LiebschM. & HalleW. (1999). Determination of the starting dose for acute oral toxicity (LD50) testing in the up and down procedure (UDP) from cytotoxicity data. ATLA27, 957–966.
20.
Anon. (2006). Peer Review Panel Report: The Use of In Vitro Basal Cytotoxicity Test Methods For Estimating Starting Doses For Acute Oral Systemic Toxicity Testing. Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) National Toxicology Program (NTP) Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM), 62pp. Research Triangle Park, NC, USA:NIEHS. Available at: http://iccvam.niehs.nih.gov/docs/acutetox_docs/ATpanelrpt06/ATpanelrpt.pdf (Accessed 18.12.07).
21.
HalleW. (2003). The registry of cytotoxicity: toxicity testing in cell cultures to predict acute toxicity (LD50) and to reduce testing in animals. ATLA31, 89–198.
22.
BlaauboerB.J., HermensJ. & van EijkerenJ. (2006). Estimating acute toxicity based on in vitro cytotoxicity: role of biokinetic modelling. ALTEX23 Special Issue, 250–253.
23.
ClemedsonC., Nordin-AnderssonM., BjerregaardH.F., ClausenJ., ForsbyA., GustafssonH., HanssonU., IsomaaB., JørgensenC., KolmanA., KotovaN., KrauseG., KristenU., KurppaK., RomertL. & ScheersE. (2002). Development of an in vitro test battery for the estimation of acute human systemic toxicity: an outline of the EDIT project. ATLA30, 313–321.
24.
CoeckeS., AhrH., BlaauboerB.J., BremerS., CasatiS., CastellJ., CombesR., CorviR., CrespiC.L., CunninghamM.L., ElautG., ElettiB., FreidigA., GennariG., Ghersi-EgeaJ-F., GuillouzoA., HartungT., HoetP., Ingelman-SundbergM., MunnS., JanssensW., LadstetterB., LeahyD., LongA., MeneguzA., MonshouwerM., MorathS., NagelkerkeF., PelkonenO., PontiJ., PrietoP., RichertL., SabbioniE., SchaackB., SteilingW., TestaiE., VericatJ-A. & WorthA. (2006). Metabolism: A bottleneck in in vitro toxicological test development. The report and recommendations of ECVAM Workshop 54. ATLA34, 49–84.
25.
ClemedsonC., BlaauboerB., CastellJ., PrietoP., RisteliL., VericatJ-A. & WendelA. (2006). ACute-Tox — Optimation and pre-validation of an in vitro test strategy for predicting human acute toxicity. ALTEX23 Special Issue, 254–258.
26.
PessinaA., AlbellaB., BayoM., BuerenJ., BrantomP., CasatiS., CroeraC., ParchmentR., Parent-MassinD., SchoeteresG., SibiriY., van den HeuvelR. & GribaldoL. (2002). In vitro tests for haematotoxicity: Prediction of drug-induced myelosuppression by the CFU–GM assay. ATLA30Suppl. 2, 75–79.
27.
PessinaA., MalerbaF. & GribaldoL. (2005). Hematotoxicity testing by cell clonogenic assay in drug development and preclinical trials. Current Pharmaceutical Design11, 1055–1065.
28.
ESAC (2006). Statement on the application of the CFU–GM assay for predicting acute neutropenia in humans. ATLA35, 201. Also available at: http://ecvam.jrc.it/index.htm (Accessed 18.12.07).
29.
GennariA., van den BergeC., CasatiS., CastellJ., ClemedsonC., CoeckeS., ColomboA., CurrenR., Dal NegroG., GoldbergA., GosmoreC., HartungT., LangezaalI., LessigiarskaI., MassW., MangelsdorfI., ParchmentR., PrietoP., SintesJ.R., RyanM., SchmuckG., StitzelK., StokesW., VericatJ-A. & GribaldoL. (2004). Strategies to replace in vivo acute systemic toxicity testing. The report and recommendations of ECVAM Workshop 50. ATLA32, 437–459.
30.
ZuccoF., BattoA-F., BisesG., ChambazJ., ChiusoloA., ConsalvoR., CrossH., Dal NegroG., De AngelisI., FabreG., GuillouF., HoffmannS., LaplancheL., MorelE., Pinçon-RaymondM., PrietoP., TurcoL., RanaldiG., RoussetM., SambuyY., ScarinoM.L., TorreillesF. & StammatiA. (2005). An inter-laboratory study to evaluate the effects of medium composition on differentiation and barrier function of Caco-2 cell lines. ATLA33, 603–618.
31.
CombesR., BallsM., IllingP., BhogalN., DaleJ., DuvéG., FeronV., GrindonC., GüldenM., LoizouG., PristonR. & WestmorelandC. (2006). Possibilities for a new approach to chemicals risk assessment — The Report of a FRAME Workshop. ATLA34, 621–649.
32.
BlaauboerB.J. (2002). The applicability of in vitro-derived data in hazard identification and characterisation of chemicals. Environmental Toxicology & Pharmacology11, 213–225.
33.
BlaauboerB.J. (2003). The integration of data on physico–chemical properties, in vitro-derived toxicity data and physiologically-based kinetic and dynamic modelling as a tool in hazard and risk assessment: A commentary. Toxicology Letters138, 161–171.
34.
LoizouG. (2004). Balancing benefit against risk. In Public Service Review: Department of Trade and Industry, pp. 57–59. Newcastle-under-Lyme, Staffordshire, UK:PSCA International Ltd. Available at: www.publicservice.co.uk/pdf/dti/6/DTI6%200601%20Georg…Loizou%20ATL.pdf (Accessed 18.12.07).
35.
LessigiarskaI., WorthA.P. & NetzevaT.I. (2005). Comparative Review of QSARs for Acute Toxicity, European Commission Report No. EUR 21559 EN, 154pp. Ispra, Italy:European Commission Joint Research Centre. Available at: http://ecb.jrc.it/qsar/publications/ (Accessed 18.12.07).
36.
DevillersJ. (2004). Prediction of mammalian toxicity of organophosphorus pesticides from QSTR modelling. SAR & QSAR in Environmental Research15, 501–510.
37.
WangG.L. & BaiN.B. (1998). Structure–activity relationships for rat and mouse LD50 of miscellaneous alcohols. Chemosphere36, 1475–1483.
38.
DuffyJ.C. (2004). Prediction of pharmacokinetic parameters in drug design and toxicology. In Predicting Chemical Toxicity and Fate (ed. CroninM.T.D. & LivingstoneD.J.), pp. 229–261. Boca Raton, FL, USA:CRC Press.
39.
EganW.J., WaltersW.P. & MurckoM.A. (2002). Guiding molecules towards drug-likeness. Current Opinion in Drug Discovery & Development5, 540–549.
40.
EkinsS., BoulangerB., SwaanP.W. & HupceyM.A.Z. (2000). Towards a new age of virtual ADME/TOX and multidimensional drug discovery. Molecular Diversity5, 255–275.
41.
EkinsS., WallerC.L., SwaanP.W., CrucianiG., WrightonS.A. & WikelJ.H. (2000). Progress in predicting human ADME parametersin silico. Journal of Pharmacological & Toxicological Methods44, 251–272.
42.
WinklerD.A. (2004). Neural networks in ADME and toxicity prediction. Drugs of the Future29, 1043–1057.
43.
Van de WaterbeemdH. (2002). High-throughput and in silico techniques in drug metabolism and pharmacokinetics. Current Opinion in Drug Discovery & Development5, 33–43.
44.
LewisD.F.V. (1999). Frontier orbitals in chemical and biological activity: Quantitative relationships and mechanistic implications. Drug Metabolism Reviews31, 755–816.
45.
HanschC., MekapatiS.B., KurupA. & VermaR.P. (2004). QSAR of cytochrome P450. Drug Metabolism Reviews36, 105–156.
46.
De GraafC., VermeulenN.P.E. & FeenstraK.A. (2005). Cytochrome P450 in silico: an integrative modeling approach. Journal of Medicinal Chemistry48, 2725–2755.
47.
De GrootM.J., KirtonS.B. & SutcliffeM.J. (2004). In silico methods for predicting ligand binding determinants of cytochromes P450. Current Topics in Medicinal Chemistry4, 1803–1824.
48.
KulkarniS.A., ZhuJ. & BlechingerS. (2005). In silico techniques for the study and prediction of xenobiotic metabolism: A review. Xenobiotica35, 955–973.
49.
MaddenJ.C. & CroninM.T.D. (2006). Structure-based methods for the prediction of drug metabolism. Expert Opinion on Drug Metabolism & Toxicology2, 545–557.
50.
MekenyanO.G., DimitrovS.D., PavlovT.S. & VeithG.D. (2004). A systematic approach to simulating metabolism in computational toxicology. I: The TIMES heuristic modelling framework. Current Pharmaceutical Design10, 1273–1293.
51.
LewisD.F.V. (2001). COMPACT: a structural approach to the modelling of cytochromes P450 and their interactions with xenobiotics. Journal of Chemical Technology & Biotechnology76, 237–244.
52.
MossG.P., DeardenJ.C., PatelH. & CroninM.T.D. (2002). Quantitative structure–permeability relationships (QSPRs) for percutaneous absorption. Toxicology in Vitro16, 299–317.
53.
BasakS.C., MillsD., HawkinsD.M. & El-MasriH.A. (2002). Prediction of tissue–air partition coefficients: A comparison of structure-based and property-based methods. SAR & QSAR in Environmental Research13, 649–665.
EganW.J. & LauriG. (2002). Prediction of intestinal permeability. Advanced Drug Delivery Reviews54, 273–289.
56.
SeibertH., GüldenM. & VossJ-U. (1994). An in vitro toxicity testing strategy for the classification and labelling of chemicals according to their potential acute lethal potency. Toxicology in Vitro8, 847–850.
57.
SeibertH., BallsM., FentemJ.H., BianchiV., ClothierR.H., DierickxP.J., EkwallB., GarleM., Gómez-LechónM.J., GribaldoL., GüldenM., LiebschM., RasmussenE., RoguetR., ShrivastavaR. & WalumE. (1996). Acute toxicity testing in vitro and the classification and labelling of chemicals. The report and recommendations of ECVAM Workshop 16. ATLA24, 499–510.
58.
BlaauboerB.J., BarrattM.D. & HoustonJ.B. (1999). The integrated use of alternative methods in toxicological risk evaluation. ECVAM integrated testing strategies task force report 1. ATLA27, 229–237.
59.
De JonghJ., ForsbyA., HoustonJ.B., BeckmanM., CombesR. & BlaauboerB.J. (1999). An integrated approach to the prediction of systemic toxicity using computer-based biokinetic models and biological in vitro test methods: overview of a prevalidation study based on the ECITTS project. Toxicology in Vitro13, 549–554.
60.
Gubbels-van HalW.M.L.G.BlaauboerB.J., BarentsenH.M., HoitinkM.A., MeertsI.A.T.M. & van der HoevenJ.C.M. (2005). An alternative approach for the safety evaluation of new and existing chemicals, an exercise in integrated testing. Regulatory Toxicology & Pharmacology42, 284–295.
61.
WorthA.P. & BallsM. (eds) (2002). Alternative (non-animal) methods for chemicals testing: Current status and future prospects. A report prepared by ECVAM and the ECVAM Working Group on Chemicals. ATLA30Suppl. 1, 125pp.
62.
LangleyG. (2001). The Way Forward — Action to End Animal Toxicity Testing, 36pp. London, UK:British Union for the Abolition of Vivisection (BUAV).
63.
ZbindenG. & Flury-RoversyM. (1981). Significance of the LD50 test for the toxicological evaluation of chemical substances. Archives of Toxicology47, 77–99.
64.
GrayA.C., MaltonJ. & ClothierR.H. (2004). The development of a standardised protocol to measure squamous differentiation in stratified epithelia, by using the fluorescein cadaverine incorporation technique. ATLA32, 91–100.
65.
PhillipsJ., KlussB., RichterA. & MasseyE.D. (2005). Exposure of bronchial epithelial cells to whole cigarette smoke: Assessment of cellular responses. ATLA33, 239–248.
66.
ThorntonD.J., GrayT., NettesheimP., HowardM., Seok KooJ., & SheehanJ.K. (2000). Characterization of mucins from cultured normal human tracheobronchial epithelial cells. American Journal of Physiology: Lung Cellular & Molecular Physiology278, L1118–L1128.
67.
ColvinV.L. (2003). The potential environmental impact of engineered nanomaterials. Nature Biotechnology21, 1166–1170.
68.
HussainS.M., HessK.L., GearhartJ.M., GeissK.T. & SchlagerJ.J. (2005). In vitro toxicity of nanoparticles in BRL 3A rat liver cells. Toxicology in Vitro19, 975–983.
69.
PoggeA. & SlikkerW.Jr (2004). Neuroimaging: new approaches for neurotoxicology. Neurotoxicology25, 525–531.
70.
ZhaoM., YangM., BaranovE., WangX., PenmanS., MoossaA.R. & HoffmanR.M. (2001). Spatial–temporal imaging of bacterial infection and antibiotic response in intact animals. Proceedings of the National Academy of Sciences of the USA98, 9814–9818.
DoeJ.E., BoobisA.R., BlackerA., DellarcoV., DoerrerN.G., FranklinC., GoodmanJ.I., KronenbergJ.M., LewisR., McConnellE.E., MercierT., MorettoA., NolanC., PadillaS., PhangW., SoleckiR., TilburyL., van RavenzwaayB. & WolfD.C. (2006). A tiered approach to systemic toxicity testing for agricultural chemical safety assessment. Critical Reviews in Toxicology36, 37–68.
73.
CombesR., BallsM., BansilL., BarrattM., BellD., BothamP., BroadheadC., ClothierR., GeorgeE., FentemJ., JacksonM., IndansI., LaozouG., NavaratnamV., PentreathV., PhillipsB., StemplewskiH. & StewartJ. (2002). An assessment of progress in the use of alternatives in toxicity testing since the publication of the report of the Second FRAME Toxicity Committee (1991). ATLA30, 365–406.
74.
BothamP. (2004). Acute systemic toxicity — prospects for tiered testing strategies. Toxicology in Vitro18, 227–230.
75.
PurchaseI.F.H. (1996). In vitro toxicology methods in risk assessment. ATLA24, 325–331.
76.
BorenfreundE. & PuernerJ.A. (1985). Toxicity determined in vitro by morphological alterations and Neutral Red absorption. Toxicology Letters24, 119–124.
77.
RiddellR.J., ClothierR.H. & BallsM. (1986). An evaluation of three in vitro cytotoxicity assays. Food & Chemical Toxicology24, 469–471.
78.
LiebschM. & SpielmannH. (1995). Balb/c 3T3 cytotoxicity test. In Methods and Molecular Biology, Vol. 43: In Vitro Toxicity Testing Protocols (ed. O'HareS. & AtterwillC.K.), pp. 177–187. Totowa, NJ, USA:Humana Press.
79.
ClothierR.H., HulmeL., SmithM. & BallsM. (1987). A comparison of the in vitro cytotoxicities and acute in vivo toxicities of 59 chemicals. Molecular Toxicology1, 571–577.
80.
ClothierR.H., HulmeL., AhmedA.B., ReevesH.L., SmithM. & BallsM. (1988). In vitro cytotoxicity of 150 chemicals to 3T3-L1 cells, assessed by the FRAME Kenacid Blue Method. ATLA16, 84–95.
81.
HulmeL., ReevesH.L., ClothierR.H., SmithM. & BallsM. (1987). An assessment of two alternative methods for predicting the in vivo toxicities of metallic compounds. Molecular Toxicology1, 589–596.
82.
SmithL.M., ClothierR.H., HillidgeS. & BallsM. (1992). Modification of the FRAME Kenacid Blue method for cytotoxicity tests on volatile materials. ATLA20, 230–234.
83.
ShrivastavaR., JohnG.W., RispatG., ChevalierA. & MassinghamR. (1991). Can the in vivo maximum tolerated dose be predicted using in vitro techniques? A working hypothesis. ATLA19, 393–402.
84.
ShrivastavaR., DelomenieC., ChevalierA., JohnG., EkwallB., WalumE. & MassinghamR. (1992). Comparison of in vivo acute lethal potency and in vitro cytotoxicity of 48 chemicals. Cell Biology & Toxicology8, 157–170.
85.
PfallerW. & TroppmairE. (2000). Renal transepithelial resistance (TER) and paracellular permeability (PCP) are reliable endpoints to screen for nephrotoxicity. In Progress in the Reduction Refinement and Replacement of Animal Experimentation (Volume 1) (ed. BallsM., van ZellerA.M. & HalderM.), pp. 291–304. Amsterdam, The Netherlands:Elsevier Science B.V.
86.
DuffT., CarterS., FeldmanG., McEwanG., PfallerW., RhodesP., RyanM. & HawksworthG. (2002). Transepithelial resistance and inulin permeability as endpoints in in vitro nephrotoxicity testing. ATLA30Suppl. 2, 53–59.
87.
DierickxP. (1989). Cytotoxicity testing of 114 compounds by the determination of the protein content in HepG2 cell cultures. Toxicology in Vitro3, 189–193.
88.
KangasL., GronroosM. & NieminenA.L. (1984). Bioluminescence of cellular ATP: a new method for evaluating cytotoxicity agentsin vitro. Medical Biology62, 338–343.
89.
WakuriS., IzumiJ., SasakiK., TanakaN. & OnoH. (1993). Cytotoxicity study of 32 MEIC chemicals by colony formation and ATP assays. Toxicology in Vitro7, 517–521.
90.
EkwallB. & SandströmB. (1978). Combined toxicity to HeLa cells of 30 drug pairs, studied by a two-dimensional microtitre method. Toxicology Letters2, 285–292.
91.
EkwallB. (1980). Preliminary studies on the validity of in vitro measurement of drug toxicity using HeLa cells. II: Drug toxicity in the MIT-24 system compared with mouse and human lethal dosage of 53 drugs. Toxicology Letters5, 309–317.
92.
EkwallB. & JohansonA. (1980). Preliminary studies on the validity of in vitro measurement of drug toxicity using HeLa cells. I: Comparative in vitro cytotoxicity of 27 drugs. Toxicology Letters5, 299–307.
93.
SkaanildM.T. & ClausenJ. (1989). Estimation of LC50 values by assay of lactate dehydrogenase and DNA redistribution in human lymphocyte cultures. ATLA16, 293–296.
94.
KempR.B., MeredithR.W.J., GambleS. & FrostM. (1983). A rapid cell culture technique for assessing the toxicity of detergent-based products in vitro as a possible screen for eye irritancyin vivo. Cytobiosystems36, 153–159.
95.
KempR.B., CrossD.M. & MeredithR.W.J. (1988). Comparison of cell death and adenosine triphosphate as indicators of acute toxicityin vitro. Xenobiotica18, 633–639.
96.
WalumE. & PetersonA. (1982). Tritiated 2-deoxy-D-glucose as a probe for cell membrane permeability studies. Analytical Biochemistry120, 8–11.
97.
WalumE. & MarchnerH. (1983). Effects of mercuric chloride on the membrane integrity of cultured cell lines. Toxicology Letters18, 89–95.
GarleM.J., KhanJ., & FryJ.R. (1988). Depletion of glutathione by the hepatotoxins paracetamol and bromobenzene, and their non-hepatotoxic analogues, in a fortified liver microsomal system. Toxicology in Vitro2, 247–252.
100.
FerroM., BassiA.M. & NanniG. (1988). Hepatoma cell cultures as in vitro models for the hepatotoxicity of xenobiotics. ATLA16, 32–37.
101.
FerroM., BassiA.M., PencoS., PianaS., UsiglioD. & NanniG. (1992). Comparative assessment of the cytotoxic effects of different xenobiotics in three hepatoma cell lines. Arzneimittel-Forschung/Drug Research442, 1053–1057.
102.
BassiA.M., PianaS., PencoS., BoscoO., BrenciS. & FerroM. (1991). Use of an established cell line in the evaluation of the cytotoxic effects of various chemicals. Bollettino della Societa Italiana di Biologia Sperimentale67, 809–816.
103.
BassiA.M., BoscoO., BrenciS., AdamoD., PencoS., PianaS., FerroM. & NanniG. (1993). Evaluation of the cytotoxicity of the first 20 MEIC chemicals in two hepatoma cell lines with different xenobiotic metabolism capacities. ATLA21, 65–72.
104.
SippelH. & EstlerC-J. (1990). Comparative evaluation of hepatotoxic side effects of various new trypanocidal diamidines in rat hepatocytes and mice. Arzneimittel-Forschung/Drug Research40, 290–293.
105.
HolzerC. & MaierP. (1987). DNA and protein contents of hepatocytes in primary cultures monitored by flow cytometry: effect of phenobarbital and dimethylsulphoxide. Toxicology in Vitro1, 203–213.
106.
HolzerC. & MaierP. (1987). Maintenance of periportal and pericentral oxygen tensions in primary rat hepatocyte cultures: influence on cellular DNA and protein content monitored by flow cytometry. Journal of Cellular Physiology133, 297–304.
107.
MaierP., SchawalderH. & ElsnerJ. (1991). Single cell analysis in toxicity testing: the mitogenic activity of thioacetamide in cultured rat hepatocytes analyzed by DNA/protein flow cytometry. Archives of Toxicology65, 454–464.
108.
WormserU., Ben ZakineS., StivelbandE., EisenO. & NyskaA. (1990). The liver slice system: A rapid in vitro acute toxicity test for primary screening of hepatotoxic agents. Toxicology in Vitro4, 783–789.
109.
WormserU., Ben ZakineS. & NyskaA. (1990). Cadmium-induced metallothionein synthesis in the rat liver slice system. Toxicology in Vitro4, 791–794.
110.
ParzefallW., MonschauP. & Schulte-HermannR. (1989). Induction by cyproterone acetate of DNA synthesis and mitosis in primary cultures of adult rat hepatocytes in serum free medium. Archives of Toxicology63, 456–446.
111.
Gómez-LechónM.J., LópezP., DonatoT., MontoyaA., Larrauri GiménezP., TrullenqueR., FabraR. & CastellJ.V. (1990). Culture of human hepatocytes from small surgical biopsies. Biochemical characterization and comparison within vivo. In Vitro Cellular Development Biology26, 67–74.
112.
DoehmerJ. (1993). V79 Chinese hamster cells genetically engineered for cytochrome P450 and their use in mutagenicity and metabolism studies. Toxicology82, 105–118.
113.
JensenK.G., LoftS., DoehmerJ. & PoulsenH.E. (1993). Metabolism of phenacetin in V79 Chinese hamster cell cultures expressing rat liver cytochrome P450 1A2 compared to isolated rat hepatocytes. Biochemical Pharmacology45, 1171–1173.
114.
RileyR.J., RobertsP., ColemanM.D., KitteringhamN.R. & ParkB.K. (1990). Bioactivation of dapsone to a cytotoxic metabolite: in vitro use of a novel two compartment system which contains human tissues. British Journal of Clinical Pharmacology30, 417–426.
115.
TingleM.D., ColemanM.D. & ParkB.K. (1990). An investigation of the role of metabolism in dapsone-induced methaemoglobinaemia using a two compartment in vitro test system. British Journal of Clinical Pharmacology30, 829–838.
116.
GüldenM., SeibertH., VossJ-U. & WassermannO. (1992). Animal cells in vitro as a supplement or alternative to animals in acute toxicity testing. Schriftenreihe des Instituts für Toxikologie der Universität Kiel Heft21, 1–185.
117.
GüldenM., SeibertH. & VossJ-U. (1994). In vitro toxicity screening using cultured rat skeletal muscle cells. II: Agents affecting excitable membranes. Toxicology in Vitro8, 197–206.
118.
GüldenM., SeibertH. & VossJ-U. (1994). Inclusion of physicochemical data in quantitative comparisons of in vitro and in vivo toxic potencies. ATLA22, 185–192.
119.
GüldenM., SeibertH. & VossJ-U. (1994). The use of cultured skeletal muscle cells in testing for acute systemic toxicity. Toxicology in Vitro8, 779–782.
120.
AtterwillC.K. (1989). Brain reaggregate cultures in neurotoxicological investigations: studies with cholinergic neurotoxins. Scandinavian Cell Toxicology Congress 1. ATLA16, 221–230.
121.
BachP.H. (1989). The detection of chemically induced renal injury, the cascade of degenerative morphological and functional changes that follow the primary nephrotoxic insult and the evaluation of these changes by in vitro methods. Toxicology Letters46, 237–250.
122.
KwizeraE.N., WilksM.F. & BachP.H. (1990). Effects of aminoglycosides on the incorporation of amino acids and on fatty acid oxidation in freshly isolated rat renal proximal tubules. In Vitro Toxicology3, 243–253.
123.
WilksM.F., KwizeraE.N. & BachP.H. (1990). Assessment of heavy metal nephrotoxicity in vitro using isolated rat glomeruli and proximal tubular fragments. Renal Physiology & Biochemistry13, 275–284.
124.
WilliamsP.D., LaskaD.A., TagL.K. & HottendorfG.H. (1988). Comparative toxicities of cephalosporin antibiotics in a rabbit kidney cell line (LLC-RK1). Antimicrobial Agents & Chemotherapy32, 314–318.
125.
BoogaardP.J., CommandeurJ.N.M., MulderJ.G., VermeulenN.P.E. & NagelkerkeJ.F. (1989). Toxicity of the cysteine-S-conjugates and mercapturic acids of four structurally related difluorethylenes in isolated proximal tubular cells from rat kidney: Uptake of the conjugates and activation to toxic metabolites. Biochemical Pharmacology38, 3731–3741.
126.
BoogaardP.J., MulderG.J. & NagelkerkeJ.F. (1989). Isolated proximal tubular cells from rat kidney as an in vitro model for studies on nephrotoxicity. II: Methylglucose uptake as a sensitive parameter for mechanistic studies of acute toxicity by xenobiotics. Toxicology & Applied Pharmacology101, 144–157.
127.
KesinglandK., EarlL.K., RobertsJ.C. & JonesH.B. (1991). Allylamine toxicity in embryonic myocardial myocyte reaggregate cultures. Toxicology in Vitro5, 145–156.
128.
EarlL.K., KesinglandK., DavisK.P., BrocklehurstS.R. & JonesH.B. (1992). Allylamine toxicity in embryonic myocardial myocyte reaggregate cultures: The role of extracellular metabolism by benzylamine oxidase. Toxicology in Vitro6, 405–416.
129.
SeamanC.W., ToselandC.D.N., MaileP.A., FrancisI. & WhiteD.J. (1994). A study of the cardiotoxic potential of pharmaceutical compounds on chick myocardial myocyte reaggregate cultures. Toxicology in Vitro8, 543–544.
130.
LewisI.D., RawlingT., DysonP.G., HaylockD.N., JuttnerD.N. & ToL.B. (1996). Standardization of the CFU–GM assay using hematopoietic growth factors. Journal of Hematometery5, 625–630.
131.
PessinaA. (1998). The Granulocyte Macrophage Colony-Forming Unit Assay. In Animal Cell Culture Techniques (ed. ClynesM.), pp. 217–230. New York, NY, USA:Springer-Verlag.
132.
PessinaA., AlbellaB., BuerenJ., BrantomP., CasatiS., CorraoG., GribaldoL., ParchmentR., Parent-MassinD., PiccirilloM., RioB., SacchiS., SchoetersG. & Van den HeuvelR. (2000). Method development for a prevalidation study of in vitro GM–CFU assay for predicting myelotoxicity. In Progress in the Reduction, Refinement and Replacement of Animal Experimentation (ed. BallsM., van ZellerA-M. & HalderM.), pp. 679–691. Amsterdam, The Netherlands:Elsevier Science B.V.
133.
PessinaA., AlbellaB., BuerenJ., BrantomP., CasatiS., GribaldoL., CroeraC., GagliardiG., FotiP., ParchmentR., Parent-MassinD., SibirilY., SchoetersG. & Van den HeuvelR. (2001). Prevalidation of a model for predicting acute neutropenia by colony forming unit granulocyte/macrophage (CFU–GM) assay. Toxicology in Vitro15, 729–740.
134.
PessinaA., AlbellaB., BayoB., BuerenJ., BrantomP., CasatiS., CroeraC., GagliardiG., FotiP., ParchmentR., Parent-MassinD., SchoetersG., SibirilY., Van den HeuvelR. & GribaldoL. (2003). Application of the CFU–GM assay to predict acute drug-induced neutropenia: an international blind trial to validate a prediction model for the maximum tolerated dose (MTD) of myelosuppressive xenobiotics. Toxicological Sciences75, 355–367.
CollanY., KosmaV-M., KuljuT., VäänänenI., Remola-PärssinenE., PesonenE., PuhakainenR., Rytöluoto-KärkkäinenR., ManninenR. & PasanenJ. (1988). Estimation of dust toxicity in rat alveolar macrophage cultures. In Safety Evaluation of Chemicals on Laboratory Animals, Proceedings of the Finnish–Soviet Symposium, Kuopio 20–22 May 1986 (ed. NevalainenT., VoipioH-M. & HaatajaH.), pp 105–123. Kuopio, Finland:University of Kuopio.
137.
DevereuxT.R. (1984). Alveolar type II and Clara cells: isolation and xenobiotic metabolism. Environmental Health Perspectives56, 95–101.
138.
DevereuxT.R., JonesK., BendJ., FoutsJ., StrathamC. & BoydM.R. (1982). In vitro metabolic activation of the pulmonary toxin, 4-ipomeanol, in non-ciliated bronchiolar epithelial (Clara) and alveolar type II cells isolated from rabbit lung. Journal of Pharmacology & Experimental Therapeutics220, 223–227.
139.
BondJ.A., MitchellC.E. & LiA.P. (1983). Metabolism and macromolecular covalent binding of benzo[a]pyrene in cultured Fischer-344 rat lung type II epithelial cells. Biochemical Pharmacology32, 3771–3776.
140.
ZamoraP.O., BensonJ.M., MarshallT.C., MoklerB.V., LiA.P., DahlA.R., BrooksA.L. & McClellanR.O. (1983). Cytotoxicity and mutagenicity of vapor phase environmental pollutants in rat lung epithelial cells and Chinese hamster ovary cells grown on collagen gels. Journal of Toxicology & Environmental Health12, 27–38.
141.
GanL.S.L. & ThakkerD.R. (1997). Applications of the Caco-2 model in the design and development of orally active drugs: Elucidation of biochemical and physical barriers posed by the intestinal epithelium. Advanced Drug Delivery Reviews23, 77–98.
142.
KansyM., SennerF. & GubernatorK. (1998). Physicochemical high throughput screening: parallel artificial membrane permeation assay in the description of passive absorption processes. Journal of Medicinal Chemistry41, 1007–1010.
143.
HukkanenJ., LassilaA., PäivärintaK., ValanneS., SarpoS., HakkolaJ., PelkonenO. & RaunioH. (2000). Induction and regulation of xenobiotics metabolising cytochrome P450s in human A549 lung adenocarcinoma cells. American Journal of Respiratory Cellular Molecular Biology22, 360–366.
144.
DimovaS., BrewsterM.E., NoppeM., JorissenM. & AugustjinsP. (2005). The use of human nasal in vitro cell systems during drug discovery and development. Toxicology in Vitro19, 107–122.
145.
ReichlS., BednarzJ. & Muller-GoymannC.C. (2004). Human corneal equivalent as cell culture model for in vitro drug permeation studies. British Journal of Ophthalmology88, 560–565.
146.
HeringaM.B., SchreursR.H.M.M., BusserF., Van Der SaagP.T., Van Der BurgB. & HermensJ.L.M. (2004). Toward more useful in vitro toxicity data with measured free concentrations. Environmental Science & Technology38, 6263–6270.
147.
GarbergP. (1998). In vitro models of the blood–brain barrier. ATLA26, 821–847.
148.
TahtiH., NevalaH. & ToimelaT. (2003). Refining in vitro neurotoxicity testing — The development of blood–brain barrier models. ATLA31, 273–276.
149.
ViravaidyaK. & ShulerM.L. (2002). Prediction of naphthalene bioaccumulation using an adipocyte cell line model. Biotechnology Progress18, 174–181.
150.
CanovaN., KmonicovaE., LincovaD., VitekL. & FarghaliH. (2004). Evaluation of a flat membrane hepatocyte bioreactor for pharmacotoxicological applications: Evidence that inhibition of spontaneously produced nitric oxide improves cell functionality. ATLA32, 25–35.
151.
LeeM.Y., ParkC.B., DordickJ.S. & ClarkD.S. (2005). Metabolizing enzyme toxicology assay chip (MetaChip) for high-throughput microscale toxicity analyses. Proceedings of the National Academy of Sciences of the USA102, 983–987.
152.
ElautG., TörökG., PapeleuP., Van HaeckeT., LausG., TourwéD. & RogiersV. (2004). Rat hepatocyte suspensions as a suitable in vitro model for studying the biotransformation of histone deacetylase inhibitors. ATLA32Suppl. 1, 105–112.
153.
KrebsfaengerN., MuedterT.E., ZangerU.M., EichelbaumM.F. & DoehmerJ. (2003). V79 Chinese hamster cells genetically engineered for polymorphic cytochrome P450 2D6 and their predictive value for humans. ALTEX20, 143–154.
154.
BullS., LangezaalI., ClothierR. & CoeckeS. (2001). A genetically engineered cell-based system for detecting metabolism-mediated toxicity. ATLA29, 703–716.
155.
TownsendA.J., KablerS.L., DoehmerJ. & MorrowC.S. (2002). Modelling the metabolic competency of glutathione S-transferases using genetically modified cell lines. Toxicology181–182, 265–269.
156.
PerazaM.A., CarterD.E. & GandolfiA.J. (2003). Toxicity and metabolism of subcytotoxic inorganic arsenic in human renal proximal tubule epithelial cells (HK-2). Cell Biology & Toxicology19, 253–264.
