In recent years, plasma activated medicine using non-thermal atmospheric-pressure plasma (NTAPP) has attracted great interest for chemotherapy. In this study, we aim to develop a chemotherapy system with reactive species generated in a plasma activated medium (PAM) for PC12 treatment. We observe the morphology change of PC12 cells and evaluate the effectiveness of PAM and the reactive species on axonal-extension enhancement.
First, we measured the amount of reactive species, such as H2O2, NO2, and NO3 in PAM. Second, we confirmed the stimulation effect of PAM on PC12 by measuring the axonal extension length of a 72-h culture after PAM stimulation. Experiments were conducted under 20 conditions to find an optimal condition. Using the grid method, significant axonal extension was confirmed. Next, the optimal conditions to promote PC12 axonal-extension was determined using the response surface method. Promotion of axonal extension was not confirmed in the cell culture using only H2O2. As a result, we presumed that the enhancement of axonal-extension was induced by the coupling effects of NO2, NO3 and H2O2, which are active species produced in PAM. Finally, we were able to declare that PAM exhibits a selective cell activation property for the chemotherapy of the central nervous system.