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In this study, an axonal extension enhancement system was developed using plasma-activated medium (PAM), which was generated through a nonthermal atmospheric-pressure plasma-bubbling system. Until recently, a number of regeneration methods for treatment of an injured central nervous system (CNS) have been proposed and examined. However, problems remain regarding quick recovery. It is also strongly required to establish an efficacious regeneration method for an injured CNS. Therefore, this study aimed to enhance the nerve axonal extension using PAM. The effectiveness of PAM in enhancing axonal extension was evaluated. First, the amounts of reactive species, such as H2O2, NO2, and NO3, in PAM were measured. Second, the stimulating effect of PAM was investigated by morphologically evaluating the PC12 cells after PAM stimulation. Experiments were conducted under 20 conditions to determine an optimal condition. Furthermore, the mechanism of enhancing PC12 neurite extension using PAM was elucidated. It was found that the Ca2+ concentration in PC12 cells was increased upon PAM stimulation and that Erk and CREB were activated, which enhanced the neurite extension. The results show that the PAM exhibited a selective cell-activation property for the treatment of CNS injuries.

Enhancement of PC12 Neurite Extension via Plasma-Activated Medium by Nonthermal Atmospheric-Pressure Plasma-Bubbling System