Validation of in vitro tests in neurotoxicology

By Campbell, I.C. - Fletcher, L. - Grant, P.A.A. - Abdulla, E.M.

The major challenges in neurotoxicity testing relate to the complexity of the nervous system, the diversity of cell types involved, and the level of integration in the mammalian nervous system. In addition, compounds which have selective pharmacological effects as receptor agonists/antagonists (for example, strychnine) may be neurotoxic at non-cytotoxic concentrations. Tests should answer the following questions: a) when is an effect toxic?, b) when is a substance to be considered toxic? and C) is the long-term risk assessment valid? The two major strategies used in the development of in vitro neurotoxicity tests are: mechanistic, in which an attempt is made to elucidate the biochemical processes involved in neurotoxicity, and disease based, which could ultimately be the most useful strategy but which is currently constrained by lack of knowledge of the aetiology of most neurological illnesses. Potential in vitro test systems which are being developed include: a) simple tests which measure the activity of an enzyme (for example, acetylcholine esterase); b) studies involving single cell type culture for example, neuroblastomas or dorsal root ganglion cells); c) complex primary coculture systems (for example, reaggregate culture); and d) combinations of these, including tiered testing and battery testing. Unfortunately, the more complex a system is, the more extensive is the characterisation needed, and, arguably, such systems will never fully mimic the intact central nervous system/peripheral nervous system (CNS/PNS). To demonstrate some of the problems inherent in neurotoxicity test development, the use of a system which involves the inhibition of outgrowth is described. The ideas are developed to include the shift toward proliferation and/or apoptosis of non-terminally differentiated neurons. The most realistic objective for the optimised, integrated and validated in vitro reductionist approach for neurotoxicological assessment is for the screening of new compounds in parallel with: a) the in vivo holistic approach (for example, to obtain pharmacokinetics and absorption/receptor-binding data); and b) quantitative structure activity relationships (QSARs). This is necessarily a selective review, and more details of methodologies and strategies are presented in other publications.