Glutamic acid (GA) content was measured in cultured Hep G2 cells, after treatment of the cells with test compounds. The results with 37 chemicals were compared with their respective rabbit eye irritation data, of which 17 were determined according to the OECD test, and the other 20 in range-finding studies. The chemicals were mainly organic solvents (alcohols, esters, amines, acids and others). The xenobiotics were applied to the cells for 4 hours at 5 different concentrations. The cells were then incubated for 15 minutes with tritiated GA. GA uptake inhibition was measured by liquid scintillation counting, and the results were expressed as the GI50 value, which is the concentration of test compound required to induce a 50% reduction in GA uptake. A linear correlation coefficient r = 0.66 was found between the log GI50 and the mean corneal opacity scores. This value is comparable to that obtained in total protein and uridine uptake inhibition studies. However, r = 0.81 was found when the log GI50 was compared with range-finding scores, indicating that a closer relationship exists between cytotoxicity and the latter.
NardoneJ.M. (1980). The interface of toxicology and tissue culture and reflections on the carnation test.Toxicology17, 105–111.
2.
AcostaD., AnuforoD.C., and SmithR.V. (1980). Cytotoxicity of acetaminophen and papaverine in primary cultures of rat hepatocytes.Toxicology and Applied Pharmacology53, 306–314.
3.
HoldenH.T., LichterW., and SigelM.M. (1973). Quantitative methods for measuring cell growth and death. In Tissue Culture: Methods and Applications (ed. KruseP., PattersonM.K.), pp. 408–411. New York: Academic Press.
4.
ThelestamM., and MollbyR. (1976). Cytotoxic effects on the plasma membrane of human diploid fibroblasts, a comparative study of leakage tests.Medical Biology54, 39–49.
5.
ShopsisC., and SatheS. (1984). Uridine uptake inhibition as a cytotoxicity test: correlations with the Draize test.Toxicology29, 195–206.
6.
DierickxP.J., and MartensM. (1986). Validation of the uridine uptake inhibition assay in cultured human hepatoma cells.Belgisch Archief voor Sociale Geneeskunde, Hygiëne, Arbeidsgeneeskunde en Geriatrische Geneeskunde44, 470–477.
7.
GarrettN.E., CampbellJ.A., StackH.F., WatersM.D., and LewtasJ. (1981). The utilization of the rabbit alveolar macrophage and Chinese hamster ovary cell for evaluation of the toxicity of particulate materials.Environmental Research24, 324–330.
8.
BorenfreundE., and BorreroO. (1984). In vitro cytotoxicity assays: potential alternatives to the Draize ocular irritancy test.Cell Biology and Toxicology1, 55–65.
9.
North-RootH., YackovichF., DemetruliasJ., GaculaM., and HeinzeJ.E. (1982). Evaluation of an in vitro cell toxicity test using rabbit corneal cells to predict the eye-irritation potential of surfactants.Toxicology Letters14, 207–212.
10.
BorenfreundE., and PuernerJ.A. (1985). Toxicity determined in vitro by morphological alterations and neutral red absorption.Toxicology Letters24, 119–124.
11.
DillinghamE.O., MastR.W., BassG.E., and AutianJ. (1973). Toxicity of methyl- and halogen-substituted alcohols in tissue culture relative to structure-activity models and acute toxicity in mice.Journal of Pharmaceutical Sciences62, 22–30.
12.
BallsM., and BridgesJ.W. (1984). The FRAME research programme on in vitro toxicology. In Acute Toxicity Testing: Alternative Approaches (ed. GoldbergA.M.), pp. 61–80. New York: Mary Ann Liebert.
13.
DierickxP.J. (1989). Cytotoxicity testing of 114 compounds by the determination of the protein content in Hep G2 cell cultures.Toxicology In Vitro, in press.
14.
JacobsG.A., DierickxP.J., and MartensM.A. (1988). Evaluation of the in vitro uridine uptake inhibition assay in comparison with the in vivo eye irritation test as prescribed by the EEC.ATLA15, 290–296.
15.
EEC (1984). Council Directive 84/449/EEC, 19 September 1984. Adaptation of Council Directive 79/831/EEC concerning the methods for the determination of toxicity. Annex V. Part B.Official Journal of the European Community (L251), 109–112.
16.
DraizeJ.H., WoodwardG., and CalveryH.O. (1944). Methods for the study of irritation and toxicity of substances applied topically to the skin and mucous membranes.Journal
17.
DenekeS.M., SteigerV., and FauburgB.L. (1987). Effect of hyperoxia on glutathione levels and glutamic acid uptake in endothelial cells.Journal of Applied Physiology63, 1966–1971.
18.
CarpenterC.P., and SmythH.F. (1946). Chemical burns of the rabbit cornea.American Journal of Ophthalmology29, 1363–1372.
19.
SmythH.F., CarpenterC.P., WeilC.S., and PozzaniU.C. (1954). Range-finding toxicity data.Archives of Industrial Hygiene and Occupational Medicine10, 61–68.
20.
SmythH.F., CarpenterC.P., WeilC.S., PozzaniU.C., and StriegelJ.A. (1962). Range-finding toxicity data: list VI.American Industrial Hygiene Association Journal23, 95–107.
21.
SmythM.F., CarpenterC.P., WeilC.S., PozzaniU.C., StriegelJ.A., and NycumJ.S. (1969). Range-finding toxicity data: list VII.American Industrial Hygiene Association Journal30, 470–476.
22.
CarpenterC.P., WeilC.S., and SmythH.F. (1974). Range-finding toxicity data: list VIII.Toxicology and Applied Pharmacology28, 313–319.
23.
SmythH.F., CarpenterC.P., and WeilC.S. (1949). Range-finding toxicity data: list III.Journal of Industrial Hygiene and Toxicology31, 60–62.
24.
SmythH.F., and CarpenterC.P. (1948). Further experience with the range-finding test in the industrial toxicology laboratory.Journal of Industrial Hygiene and Toxicology30, 63–68.
25.
SmythH.F., and CarpenterC.P. (1944). The place of the range-finding test in the industrial laboratory.Journal of Industrial Hygiene and Toxicology26, 269–273.
26.
SmythH.F., CarpenterC.P., and WeilC.S. (1951). Range-finding toxicity data: list IV.Archives of Industrial Hygiene and Occupational Medicine4, 119–122.
27.
MarselosM., StormS.C., and MichalopoulosG. (1987). Effect of phenobarbital and 3-methylcholanthrene on aldehyde dehydrogenase activity in cultures of Hep G2 cells and normal hepatocytes.Chemico-Biological Interactions62, 75–88.
28.
DashtiN., WolfbauerG., KorenE., KnowlesB., and AlaupovicP. (1984). Catabolism of human low density lipoproteins by human hepatoma cell line Hep G2.Biochimica et Biophysica Acta794, 373–384.
29.
KnowlesB.B., HoweC.C., and AdenD.P. (1980). Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen.Science209, 497–499.
30.
DierickxP.J. (1987). Inhibition of glutathione-dependent uridine uptake in cultured human hepatoma cells.Medical Science Research15, 1349–1350.
31.
DuthieS.J., ColemanC.S., and GrantM.H. (1988). Status of reduced glutathione in the human hepatoma cell line Hep G2.Biochemical Pharmacology37, 3365–3368.
32.
TurnerL., ChoplinF., DugardP., HermensJ., JaeckhR., MarsmannM., and RobertsD. (1987). Structure-activity relationships in toxicology and ecotoxicology: an assessment.Toxicology In Vitro1, 143–171.
