Abstract
Eight inorganic metal compounds (AlCl3, Al(OH)3 gel, Al(OH)3 salt, SnCl2, ZnSO4, K2Cr2O7, CdCl2, HgCl2) were tested for their cytotoxic effect on an established hamster fibroblast line (BHK-21/C13) in vitro using a cell detachment assay and two different growth assays, the cloning effieciency and the cell number after 2 days subconfluent culture as parameters. The test conditions for these assays were optimized, including incubation period, application of test substance, growth conditions and data analysis. Aluminum, zinc and tin compounds showed low cytotoxic effects when compared to potassium, cadmium and mercuric compounds. Potassium dichromate was highly toxic in both growth assays (0.0001–0.01 mM, with a clear dependency on the incubation time), whereas it proved to be only slightly toxic in the detachment assay (0.1–5 mM). Cadmium and mercuric chlorides were the most toxic compounds in the growth (0.00001–0.001 mM) and the cell detachment assays (0.01–0.1 mM). Variable incubation periods barely affected the cytotoxicity of mercuric chloride. Ranking of these cytotoxicity data was found to be identical to the ranking of LD50 values (oral, rat) as well as to the ranking according to threshold limit values for human workroom environment, and of human eye irritation data.
Similar content being viewed by others
Abbreviations
- HK-21/C13:
-
Baby, hamster kidney fibroblasts, clone 21/C13
- CHlow :
-
lowest concentration at which a significant increase in number of detached cells was measured
- CElow :
-
lowest concentration at which a significant decrease in cloning efficiency was measured
- GIlow :
-
lowest concentration at which growth was significantly inhibited under sunconfluent culture conditions
- MEM:
-
Eagle's minimum essential medium
- TLV/TWA:
-
Threshold limit values (time weigthed average, USA) for human workroom environment
References
BADENOCH-JONES, P., TURK, J.L., and PARKER, D. (1978). The effect of some aluminium and zirkonium compounds on guinea-pig peritoneal macrophages and skin fibroblasts in culture. J. Pathol.124:51–62.
BONHAUS, D.W., MC CORMACK, K.M., MAYOR, G.H., MATTSON, J.C. and HOOK, J.B. (1980). Effects of aluminim on microtubular integrity usingin vitro andin vivo models. Toxicol. Lett.6:141–148.
CASTRANOVAS, V., BOWMAN, L., MILES, P.R., and REASOR, M.J. (1980). Toxicity of metal ions to alveolar macrophages. Amer. J. Indust. Med.1:349–357.
ELSNER, J., and KNUTTI, R. (1981). Graphical representation of the time evolution of ECG data using a computer data storage and plot package. In:The Rat Electrocardiogram in Pharmacology and Toxicology (R. Budden, D.K. Detweiler, and G. Zbinden. eds.) Pergamon Press, pp.
FARNELL, B.J., DE BONI, U., and MC LACHLAN, D.R.C. (1982). Aluminum neurotoxicity in CA1 hippocampal pyramidal neuronsin vitro. Exp. Neurol.78:241–258.
IARC Mongraphs on the evaluation of the carcinogenic risk of chemicals to humans. (1980). Some metals and metallic compounds; chromium and chromium compounds. WHO, Lyon, Vol. 23, p. 205–324.
JAKOBSEN, K., BAKKE, O., OSTGAARD, K., WHITE, L.R., EIK-NES, K.B. (1982). Effects of potassium dichromate on the cell cycles of an established human cell line (NIHK 3025). Toxicology24: 281–292.
LEVIS, A.G., BIANCHI, V., TAMINO, G., and PAGARO, B. (1978a). Cytotoxic effects of hexavalent and trivalent chromium on mammalian cellsin vitro. Br. J. Cancer37:386–396.
LEVIS, A.G., BUTTIGNOL, G., and SPONZA, G. (1978b). Effects of potassium dichromate on nuclei acid and protein synsthesis and on precursor uptake in BHK fibroblasts. Cancer Res.38:110–116.
MUMMERY, C.L., VAN DEN BRINK, S., VAN DER SAAG, P.T., AND DE LAAT, S.W. (1983a). Screening for cytotoxicity in neublastoma cells. I. Dependence of growth inhibition on the presence of serum. Toxicol. Lett.18:201–209.
MUMMERY, C.L., VAN DEN BRINK, S., VAN DER SAAG,.P.T., and DE LAAT, S.W. (1983a). Screening for cytotoxicity in neuroblastoma cells. I. Dependence of growth inhibition on the presence of serum. toxicol. Lett.18:201–209.
NIOSH (1980). Register of toxic effects of chemical substances.Washington, NIOSH Publ. Nr. 81–116.
REINHARDT, C.A., SCHAWALDER, H.P., and ZBINDEN, G. (1982). Cell detachment and cloning effeciency as parameters for cytotoxicity. Toxicology25:47–53.
REINHARDT, C.A., PELLI, D.A., and ZBINDEN, G. (3985a). Interpretation of cell toxicity data for the prediction ofin vivo irritation effects. Food Chem. Toxicol. (in press).
REINHARDT, C.A., BOSSHARD, E., and SCHLATTER, C. (eds). (1985b). Workshop preceedings on “Irritation testing of skin and mucous membranes”, April 3–5, 1984, Kartause Ittingen, Switzerland. Food Chem. Toxicol. (in press).
WALUM, E. (1982). Membrane lesions in cultured mouse neuroblastoma cells exposed to metal compounds. Toxicology25: 67–74.
WHO. (1977). Aluminium. Biological data. In:Food Additives Series Nr. 12, 14–24.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Reinhardt, C.A., Pelli, D.A. & Sandvold, M. Cell detachment and growth of fibroblasts as parameters for cytotoxicity of inorganic metal saltsin Vitro . Cell Biol Toxicol 1, 33–43 (1985). https://doi.org/10.1007/BF00717789
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00717789