Integration of in vitro neurotoxicity data with biokinetic modelling for the estimation of in vivo neurotoxicity.
Risk assessment of neurotoxicity is mainly based on in vivo exposure, followed by tests on behaviour, physiology and pathology. In this study, an attempt to estimate lowest observed neurotoxic doses after single or repeated dose exposure was performed. Differentiated human neuroblastoma SH-SY5Y cells were exposed to acrylamide, lindane, parathion, paraoxon, phenytoin, diazepam or caffeine for 72 hours. The effects on protein synthesis and intracellular free Ca2+ concentration were studied as physiological endpoints. Voltage operated Ca2+ channel function, acetylcholine receptor function and neurite degenerative effects were investigated as neurospecific endpoints for excitability, cholinergic signal transduction and axonopathy, respectively. The general cytotoxicity, determined as the total cellular protein levels after the 72 hours exposure period, was used for comparison to the specific endpoints and for estimation of acute lethality. The lowest concentration that induced 20% effect (EC20) obtained for each compound, was used as a surrogate for the lowest neurotoxic level (LOEL) at the target site in vivo. The LOELs were integrated with data on adsorption, distribution, metabolism and excretion of the compounds in physiologically-based biokinetic (PBBK) models of the rat and the lowest observed effective doses (LOEDs) were estimated for the test compounds. A good correlation was observed between the estimated LOEDs and experimental LOEDs found in literature for rat for all test compounds, except for diazepam. However, when using in vitro data from the literature on diazepam's effect on gamma-amino butyric acid (GABA)A receptor function for the estimation of LOED, the correlation between the estimated and experimental LOEDs was improved from a 10,000-fold to a 10-fold difference. Our results indicate that it is possible to estimate LOEDs by integrating in vitro toxicity data as surrogates for lowest observed target tissue levels with PBBK models, provided that some knowledge about toxic mechanisms is known.
Forsby
Anna
A
Department of Neurochemistry, Svante Arrhenius Laboratories for Natural Sciences, Stockholm University, Stockholm, Sweden. annaf@neurochem.su.se
Blaauboer
Bas
B
D020106:Acrylamide / Q000493:pharmacokinetics / Q000633:toxicity; D000818:Animals; D000927:Anticonvulsants / Q000493:pharmacokinetics / Q000633:toxicity; D001932:Brain Neoplasms / Q000473:pathology; D002110:Caffeine / Q000493:pharmacokinetics / Q000633:toxicity; D015220:Calcium Channels / Q000187:drug effects / Q000502:physiology; D000697:Central Nervous System Stimulants / Q000493:pharmacokinetics / Q000633:toxicity; D003975:Diazepam / Q000493:pharmacokinetics / Q000633:toxicity; D023381:Endpoint Determination; D006801:Humans; D006993:Hypnotics and Sedatives / Q000493:pharmacokinetics / Q000633:toxicity; D007700:Kinetics; D007928:Lethal Dose 50; D015233:Models, Statistical; D009419:Nerve Tissue Proteins / Q000096:biosynthesis; D009447:Neuroblastoma / Q000473:pathology; D020258:Neurotoxicity Syndromes / Q000473:pathology*; D010575:Pesticides / Q000493:pharmacokinetics / Q000633:toxicity; D010672:Phenytoin / Q000493:pharmacokinetics / Q000633:toxicity; D051381:Rats; D015262:Xenobiotics / Q000493:pharmacokinetics* / Q000633:toxicity*
D000927:Anticonvulsants; D015220:Calcium Channels; D000697:Central Nervous System Stimulants; D006993:Hypnotics and Sedatives; D009419:Nerve Tissue Proteins; D010575:Pesticides; D015262:Xenobiotics; D020106:Acrylamide; D002110:Caffeine; D010672:Phenytoin; D003975:Diazepam