Abstract
Local anaesthetics exhibit direct neurotoxic effects on neurones. Numerous studies have investigated the factors that may reverse this neuropathology, but the effects of glucose conditions on neuronal regeneration after lidocaine-induced injury have not been examined by observing living neurones. The present study investigated the effects of different glucose conditions on neurite length, growth cone regeneration, and cell death in dorsal root ganglia (DRG) neurones after lidocaine-induced injury in vitro.
DRG explants were isolated from chick embryos at embryonic day 8 and cultured in media containing low, normal, or high glucose concentrations (10, 25, or 40 mM) for 24 h. Tissues were exposed to lidocaine 8 mM for 1 h, then rinsed and incubated for a further 24 h. Neurite length and growth cone collapse assays were performed to assess neuronal growth and regeneration. Lactate dehydrogenase (LDH) and caspase assays were also performed to detect neuronal cell death.
Addition of lidocaine for 1 h resulted in >97% growth cone collapse and neurite destruction under all three glucose conditions. Two hours after rinsing out the lidocaine, significant reversal of growth cone collapse and neurite elongation was observed under all glucose conditions. Growth cone collapse was higher under low-glucose condition (P<0.05). High glucose negatively affected neurite length more than growth cone collapse. At 24 h, LDH release with both low- and high-glucose conditions was higher than with normal glucose (P<0.05). Low- and high-glucose conditions increased caspase 3/7 activation.
Normal glucose is optimal for neuronal recovery after lidocaine-induced injury in vitro.