This review outlines the work which has been conducted in the FRAME Alternatives Laboratory during the first ten years of the FRAME Research Programme. A number of in vitro tests, including the kenacid blue, neutral red release and fluorescein leakage assay methods, have been evaluated and have subsequently been included in validation schemes organised by the US Soap and Detergent Association, the US Cosmetic, Toiletry and Fragrance Association, the European Commission and the European Cosmetic, Toiletry and Perfumery Association, as well as in the Scandinavian multicentre evaluation of in vitro cytotoxicity testing scheme. More recently, research has been undertaken in the areas of phototoxicity, immunotoxicity, dermal toxicity and intercellular communication, in addition to investigations into fundamental mechanisms of toxicity.
BallsM., and BridgesJ.W. (1984). The FRAME Research Programme on in vitro cytotoxicology. In Alternative Methods in Toxicology, Vol. 2, Acute Toxicity Testing: Alternative Approaches (ed. GoldbergA.M.), pp. 63–79. New York: Mary Ann Liebert.
2.
BallsM., and HornerS.A. (1985). The FRAME interlaboratory programme on in vitro cytotox-icology.Food and Chemical Toxicology23, 209–213.
3.
BallsM., RiddellR.J., HornerS.A., and ClothierR.H. (1987). The FRAME approach to the development, validation and evaluation of in vitro alternative methods. In Alternative Methods in Toxicology, Vol. 5, In Vitro Methods in Toxicology: Approaches to Validation (ed. GoldbergA.M.), pp. 45–58. New York: Mary Ann Liebert.
4.
KnoxP., UphillP.F., FryJ.R., BenfordD.J., and BallsM. (1986). The FRAME multicentre project on in vitro cytotoxicity.Food and Chemical Toxicology24, 457–463.
5.
PurchaseI.F.H., FarrarD.G., and WhitakerLA. (1987). Toxicology profiles on substances used in the FRAME cytotoxicology research project.ATLA14, 184–243.
6.
BallsM., and ClothierR. (1987). Validation of alternative toxicity test systems: lessons learned and to be learned.Molecular Toxicology1, 547–559.
7.
ClothierR.H., and BallsM. (1990). Validation of alternative toxicity tests: principles, practices and cases.Toxicology in Vitro4, 692–693.
8.
BoomanK.A., CascieriT.M., DemetruliasJ., DriedgerA., GriffithJ.F., GrochoskiG.T., KongB., McCormickW.C., North-RootH., RozenM.G., and SedlakR.I. (1988). In vitro methods for estimating eye irritancy of cleaning products. Phase I: preliminary assessment.Journal of Toxicology — Cutaneous and Ocular Toxicology7, 173–185.
9.
ReaderS.J., BlackwellV., O'HaraR., ClothierR.H., GriffinG., and BallsM. (1989). A vital dye release method for assaying the short-term cytotoxic effects of chemicals and formulations.ATLA17, 28–83.
10.
ReaderS.J., BlackwellV., O'HaraR., ClothierR.H., GriffinG., and BallsM. (1990). Neutral red release from pre-loaded cells as an in vitro approach to testing for eye irritancy potential.Toxicology in Vitro4, 264–266.
11.
GettingsS.D., TealJ.J., BagleyD.M., DemetruliasJ.L., DiPasqualeL.C., HintzeK.L., RozenM.G., WeiseS.L., ChudkowskiM., MarenusK.D., PapeW.J.W., RoodyM., SchnetzingerR., SilberP.M., GlazaS.M., and KurtzP.J. (1991). The CTFA Evaluation of Alternatives Program: an evaluation of in vitro alternatives to the Draize Primary Eye Irritation Test. (Phase I) Hydro-alcoholic formulations; (Part 2) data analysis and biological significance.In Vitro Toxicology4, 247–288.
12.
Anon. (1991). Collaborative Study on the Evaluation of Alternative Methods to the Eye Irritation Test. Doc. XI/632/91 – V/E/1/131/91. Brussels: CEC.
13.
RiddellR.J., ClothierR.H., and BallsM. (1986). An evaluation of three in vitro cytotoxicity assays.Food and Chemical Toxicology24, 469–471.
14.
RiddellR.J., PanacerD.S., WildeS.M., ClothierR.H., and BallsM. (1986). The importance of exposure period and cell type in in vitro cytotoxicity tests.ATLA14, 86–92.
15.
ClothierR.H., HulmeL., AhmedA.B., ReevesH.L., SmithM., and BallsM. (1988). In vitro cytotoxicity of 150 chemicals to 3T3-L1 cells, assessed by the FRAME kenacid blue method.ATLA16, 84–95.
16.
ShawA.J., BallsM., ClothierR.H., and BatemanN.D. (1991). Predicting ocular irritancy and recovery from injury using Madin Darby canine kidney cells.Toxicology in Vitro5, 569–571.
17.
ShawA.J., ClothierR.H., and BallsM. (1990). Loss of trans-epithelial impermeability of a confluent monolayer of Madin Darby canine kidney (MDCK) cells as a determinant of ocular irritancy potential.ATLA18, 145–151.
18.
SmithL.M., ClothierR.H., HillidgeS., and BallsM. (1992). Modification of the FRAME kenacid blue method for cytotoxicity tests on volatile materials.ATLA20, 230–234.
19.
BallsM., and FentemJ.H. (1992). The use of basal cytotoxicity and target organ toxicity tests in hazard identification and risk assessment.ATLA20, 368–388.
20.
BallsM., and ClothierR.H. (1992). Cytotoxicity assays for intrinsic toxicity and irritancy. In In Vitro Methods of Toxicology (ed. WatsonR.R.), pp. 37–52. Boca Raton, Florida: CRC Press.
21.
TchaoR. (1988). Trans-epithelial permeability of fluorescein in vitro as an assay to determine eye irritants. In Alternative Methods in Toxicology, Vol. 6, Progress in In Vitro Toxicology (ed. GoldbergA.M.), pp. 271–283. New York: Mary Ann Liebert.
22.
Anon. (1993). Further steps towards replacing the Draize eye test.ATLA21, 127–128.
23.
GordonV.C., and BergmanH.C. (1987). EYTEX: an in vitro method for evaluation of ocular irritancy. In Alternative Methods in Toxicology, Vol. 5, In Vitro Methods in Toxicology: Approaches to Validation (ed. GoldbergA.M.), pp. 273–302. New York: Mary Ann Liebert.
24.
PapeW.J.W., PfannenbeckerU., and HoppeU. (1987). Validation of the red blood cell test system as an in vitro assay for the rapid screening of irritation potential of surfactants.Molecular Toxicology1, 525–536.
25.
BorenfreundE., and PuernerJ.A. (1985). Toxicity determined in vitro by morphological alterations and neutral red absorption.Toxicology Letters24, 119–124.
26.
BrunerL.H., MillerK.R., OwickiJ.C., ParceJ.W., and MuirV.C. (1991). Testing ocular irritancy in vitro with the silicon microphysiometer.Toxicology in Vitro5, 277–284.
27.
LupkeN.P. (1985). Hen's egg chorio-allantoic membrane test for irritation potential.Food and Chemical Toxicology23, 287–291.
28.
BurtonA.B.G., YorkM., and LawrenceR.S. (1981). The in vitro assessment of severe eye irritants.Food and Cosmetic Toxicology19, 471–480.
29.
PrinsenM.K., & KöeterB.W.M. (1993). Justification of the enucleated eye test with eyes of slaughterhouse animals as an alternative to the Draize eye irritation test with rabbits.Food and Chemical Toxicology31, 69–76.
30.
GautheronP., DukicM., AlixD., and SinaJ.F. (1992). Bovine corneal opacity and permeability test: an in vitro assay of ocular irritancy.Fundamental and Applied Toxicology18, 442–449.
31.
BallsM., BlaauboerB., BrusickD., FrazierJ., LambD., PembertonM., ReinhardtC., RoberfroidM., RosenkranzH., SchmidB., SpielmannH., StammatiA-L., & WalumE. (1990). Report and recommendations of the CAAT/ERGATT workshop on the validation of toxicity test procedures.ATLA18, 313–337.
32.
BagleyD.M., BothamP.A., GardnerJ.R., HollandG., KreilingR., LewisR.W., StringerD.A., and WalkerA.P. (1992). Eye irritation: reference chemicals data bank.Toxicology in Vitro6, 487–491.
33.
Anon. (1992). Technical Report No. 48: eye irritation reference chemicals data bank, 155 pp. Brussels: ECETOC.
34.
KristenV., HoppeV., and PapeW. (1993). The pollen tube growth test: a new alternative to the Draize eye irritation assay.Journal of the Society of Cosmetic Chemists44, 153–162.
35.
EkwallB., BondessonI., CastellJ.V., Gómez-LechónM.J., HellbergS., HobergJ., JoverR., PonsodaX., RomertL., StenbergK., and WalumE. (1989). Cytotoxicity evaluation of the first ten MEIC chemicals: acute lethal toxicity in man predicted by cytotoxicity in five cellular assays and by oral LD50 tests in rodents.ATLA17, 83–100.
36.
FentemJ., FryJ., GarleM.J., GuldenM., SeibertH., VossJ-U., WassermannO., PerchermeierM., and WiebelF. (1993). An international evaluation of selected in vitro toxicity test systems for predicting acute systemic toxicity. Report submitted to the CEC (Contract nos. B92/B4-3063/14086 and B92/B4-3040/14087).
37.
BallsM., AtterwillC., FentemJ., GarleM.J., and WiebelF.J. (1992). Evaluation of alternatives to animal tests for assessing the acute lethal potency and neurotoxic potential of chemicals. Report prepared for the CEC, DGXI (Contract no. B4-3081/91/8678).
38.
RohnT.T., HindsT.R., and VincenziF.F. (1993). Ion transport ATPases as targets for free-radical damage.Biochemical Pharmacology46, 525–534.
39.
LiebschM., SpielmannH., BallsM., BrandM., DöringB., DupuisJ., HolzhütterH.G., KlecakG., EplattenierH.L., LovellW.W., MaurerT., MoldenhauerF., MooreL., PapeW.J.W., PfannenbeckerU., PotthastJ., De SilvaO., SteilingW., and WillshawA. (1994). First results of the EC/COLIPA validation project on in vitro phototoxicity testing. In Alternative Methods in Toxicology, Vol. 10, In Vitro Skin Toxicology: Irritation, Phototoxicity, Sensitization (ed. RougierA., GoldbergA.M., MaibachH.I.). New York: Mary Ann Liebert.
40.
SpielmannH., BallsM., BrandM., DöringB., DupuisJ., HolzhütterH.G., KalweitS., KlecakG., EplattenierH.L., LiebschM., LovellW.W., MaurerT., MoldenhauerF., MooreL., PapeW.J.W., PfannenbeckerU., PotthastJ., De SilvaO., SteilingW., and WillshawA. (1994). EC/COLIPA project on in vitro phototoxicity testing: first results obtained with a BALB/C 3T3 phototoxicity.Toxicology in Vitro8, 793–796.
41.
SpielmannH., LovellW.W., HolzleE., JohnsonB.E., MaurerT., MirandaM.A., PapeW.J.W., SaporaO., and SladowskiD. (1994). In vitro phototoxicity testing: the report and recommendations of ECVAM workshop 2.ATLA22, 314–348.
42.
WillshawA., MooreL.J., and BallsM. (1994). In vitro alternatives for the detection of photoirritant chemicals: the EC/COLIPA trial.Toxicology in Vitro8, 723–725.
43.
SteerS., BallsM., ClothierR.H., and GordonV. (1994). The development and evaluation of in vitro tests for photoirritancy.Toxicology in Vitro8, 719–721.
44.
HornerS.A., and ShahM.K. (1984). Effects of fungizone on the in vitro cytotoxicity of other chemicals.ATLA11, 147–153.
45.
ClothierR., RobinsonT., GreengrassS., HayesV., RatcliffeJ., HornerS., and BallsM. (1985). Effects of antibiotics and organic solvents on the in vitro cytotoxicity of other chemicals.ATLA12, 203–213.
46.
HulmeL.M., ReevesH.L., ClothierR.H., SmithM., and BallsM. (1987). Assessment of two alternative methods for predicting the in vivo toxicities of metallic compounds.Molecular Toxicology1, 589–596.
47.
HornerS.A., FryJ.R., ClothierR.H., and BallsM. (1985). A comparison of two cytotoxicity assays for the detection of metabolism-mediated toxicity in vitro: a study with cyclophosphamide.Xenobiotica15, 681–686.
48.
HornerS.A., FryJ.R., ClothierR.H., and BallsM. (1986). Diethylmaleate-induced alterations in glutathione content and cytochrome P450 content and related activity in liver S9 fractions.IRCS Medical Science14, 386–387.
49.
HornerS.A., FryJ.R., ClothierR.H., and BallsM. (1987). The detection of cytotoxicity produced by generation of short-lived reactive metabolites: a study with bromobenzene.Xenobiotica17, 777–782.
50.
HornerS.A., ZuccoF., FryJ.R., ClothierR.H., and BallsM. (1987). Investigation of the cytotoxicity produced by generation of short-lived reactive metabolites in vitro: a study with paracetamol.Toxicology in Vitro1, 589–596.
51.
AtkinsonK., HulmeL., ClothierR.H., and BallsM. (1989). In vitro cytotoxicity of 24 chemicals to mouse teratocarcinoma cells.ATLA17, 34–37.
52.
HulmeL.M., AtkinsonK.A., ClothierR.H., and BallsM. (1990). The potential usefulness of a differentiating teratocarcinoma cell line in in vitro toxicity testing.Toxicology in Vitro4, 589–592.
53.
SladowskiD., SteerS., MooreL., GlassborowR., ClothierR., and BallsM. (1993). Preliminary findings of an in vitro assay based on human complement activation for the detection of chemicals with phototoxicity potential.ATLA21, 509–512.
54.
WardR.K., AgrawallaS., and ClothierR.H. (1994). Investigation of an in vitro cytotoxicity assay for prediction of skin irritation.Toxicology in Vitro8, 659–660.
55.
SladowskiD., SteerS., ClothierR., and BallsM. (1993). An improved MTT assay.Journal of Immunological Methods157, 203–207.
56.
SladowskiD., SteerS., ClothierR., and BallsM. (1994). Use of a filtration plate in the MTT assay.Toxicology in Vitro8, 739–741.
57.
SteerS., LasekW., ClothierR.H., and BallsM. (1990). An in vitro test for immunomodulators?Toxicology in Vitro4, 360–362.
58.
LasekW., SteerS., ClothierR.H., and BallsM. (1992). The stimulation of EL-4 cells to produce interleukin-2 and its potential use in immuno-cytotoxicity testing.Immunopharmacology and Immunotoxicology11, 379–396.
59.
SladowskiD.T., SteerS.J., MooreL., WillshawA., ClothierR., and BallsM. (1994). The complement activation assay: the test for immediate phototoxicity.Toxicology in Vitro8, 513–515.
60.
RheinwaldJ.G., and GreenH. (1975). Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells.Cell6, 331–344.
61.
BoyceS.T., and HamR.G. (1985). Cultivation, frozen storage and clonal growth of normal human epidermal keratinocytes in serum-free media.Journal of Tissue Culture Methods9, 83–93.
62.
FusenigN.E. (1986). Mammalian epidermal cells in culture.Biology of the Integument2, 409–443.
63.
TsaoM.C., WalthallB.J., and HamR.G. (1982). Clonal growth of normal human epidermal keratinocytes in a defined medium.Journal of Cellular Physiology110, 219–229.
64.
PonecM. (1992). In vitro cultured skin cells as alternatives to animals for skin irritancy screening.International Journal of Cosmetic Science14, 245–264.
65.
RégnierM., AsselineauD., and LenoirM.C. (1990). Human epidermis reconstructed on dermal substrates in vitro: an alternative to animals in skin pharmacology.Skin Pharmacology3, 70–85.
66.
BellE., ParenteauN., GayR., NolteC., KempP., BilboP., EksteinB., and JohnsonE. (1991). The Living Skin Equivalent: its manufacture, its organotyplc properties, and its response to irritants.Toxicology in Vitro5, 591–596.
67.
SlivkaS.R., and ZeiglerF. (1993). Use of an in vitro skin model for determining epidermal and dermal contributions to irritant response.Journal of Toxicology — Cutaneous and Ocular Toxicology12, 49–57.
68.
WardR.K., ShawB., ClothierR.H., and BallsM. (1993). Development of a human epidermal equivalent in vitro.Journal of Anatomy182, 151.
69.
FentemJ.H., CooperA., WardR.K., WillshawA., and BallsM. (1995). In vitro toxicology studies in human keratinocytes.Comments in Toxicology, in press.
70.
Anon. (1994). NO may be the answer!FRAME News35, 4.
71.
Anon. (1994). “Kind” make-up test is skin-deep.New Scientist143, 20.
72.
CooperA. (1993). Development of invitro assays to study chemical carcinogenesis and anti-cancer chemotherapeutic agents. PhD Thesis, University of NottinghamUK.
73.
CooperA., FentemJ., and BallsM. (1993). Effects of chemicals on intercellular communication in vitro.Human and Experimental Toxicology12, 557
74.
CooperA., FentemJ., and BallsM. (1994). Inhibition of intercellular communication: an in vitro assay for detecting potential tumour promoters.Toxicology in Vitro8, 535–537.
75.
ReaderS.J., ClothierR.H., YuJ., and BallsM. (1990). The in vitro effects and metabolism-mediated cytotoxicity of phorone, a glutathione-depleting agent.Toxicology in Vitro4, 509–512.
76.
KosowerN.S., KosowerE.M., NewtonG.L., and RanneyH.M. (1979). Bimane fluorescent labels: labelling of normal human red cells under physiological conditions.Proceedings of the National Academy of Sciences of the United States of America76, 3382–3386.
77.
CookJ.K., IypeS.N., and MitchellJ.R. (1991). Differential specificity of monochlorobimane for isoenzymes of human and rodent glutathione Stransferases.Cancer Research51, 1606–1611.
78.
SavilleB. (1958). A scheme for the colorimetric determination of microgram amounts of thiols.Analyst83, 670–672.
79.
BellomoG., and MirabelliF. (1988). Cytoskeleton protein thiols represent a target for quinone-induced oxidative stress.Molecular Toxicology1, 281–293.
80.
Le BelC.P., IschiropoulosH., and BondyS.C. (1992). Evaluation of the probe 2,7-dichlorofluorescin as an indicator of reactive oxygen species formation and oxidative stress.Chemical Research in Toxicology5, 227–231.
81.
LongoN., GriffinL.D., and ElsasL.J. (1991). A simple method for evaluation of Rb+ transport and Na+-K+ stoichiometry in adherent cells.American Journal of Physiology260, C1341–C1346.
82.
MoncadaS., PalmerR.M.J., and HiggsE.A. (1991). Nitric oxide: physiology and pharmacology.Pharmacological Reviews43, 109–142.
83.
ClothierR.H., MorganS.J., AtkinsonK.A., GarleM.J., and BallsM. (1994). Development of a fixed-dose approach for the fluorescein leakage test.Toxicology in Vitro8, 883–884.
84.
HubbardA.W., MooreL.J., ClothierR.H., SulleyH., and RollinK.A. (1994). The use of in vitro methodology to predict the irritancy potential of surfactants.Toxicology in Vitro8, 689–691.
85.
AtkinsonK.A., FentemJ.H., ClothierR.H., and BallsM. (1992). Alternatives to ocular irritation testing in animals.Lens and Eye Toxicity Research9, 247–258.
86.
BallsM., AtkinsonK.A., and GordonV.C. (1991). Complementation in the development, validation and use of non-animal test batteries, with particular reference to ocular irritancy.ATLA19, 429–431.
87.
ClothierR.H., HulmeL.M., SmithM., and BallsM. (1987). Comparison of the in vitro cytotoxicities and acute in vivo toxicities of 59 chemicals.Molecular Toxicology1, 571–577.
88.
BallsM., and FentemJ. (1994). The Fund for the Replacement of Animals in Medical Experiments (FRAME): 23 years of campaigning for the Three Rs. In Alternatives to Animal Testing. New Ways in the Biomedical Sciences, Trends and Progress (ed. ReinhardtC.A.), pp. 45–55. Weinheim, Germany: VCH Verlag.
