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Toxicity and integrity disruption in response to transport through the blood-brain barrier (BBB) of the organophosphates malathion and malaoxon and heavy metal lead acetate were assessed in two in vitro barrier systems. One system was constructed using bovine brain microvascular endothelial cells (BMEC), while the other system was constructed with rat brain microvascular endothelial cells (RBE4); both were cocultured with rat astrocytes. We hypothesized that these models would respond differently to neurotoxic compounds. Concentrations of malathion, malaoxon, and lead acetate between 0.01 microM and 1 mM were assessed for their capacity to cause cytotoxicity to the astrocytes and endothelial cells utilized to construct the BBB systems, with the least cytotoxic concentrations chosen for transfer assessments of neurotoxicants through the barrier systems. Concentrations of malathion at 10 microM, malaoxon at 1 microM, and lead acetate at 1 and 10 microM were selected. Lead concentrations were measured in media of the abluminal and luminal sides of both systems using graphite furnace atomic absorption at the beginning of the treatment (T0) and 14 h later (T14). Passage of organophosphate compounds was determined utilizing inhibition of acetylcholinesterase enzyme in a neuroblastoma cell line (SH-SY5Y) localized below the barrier system. Transendothelial electrical resistance was assessed as a measurement of integrity of the barrier systems, with baseline values higher with the RBE4-astrocyte system than with the BMEC-astrocyte system. Metabolic capability, as measured by esterase activity, was higher in BMECs, which were more likely to retain lead than RBE4 cells. Results suggest that differences in endothelial cell source can affect the outcome of studies on toxicant transfer through in vitro BBB systems.

Comparison of Two Blood-Brain Barrier In Vitro Systems: Cytotoxicity and Transfer Assessments of Malathion/Oxon and Lead Acetate