This study investigates the neuroprotective effects of helium-based atmospheric pressure plasma jet (APPJ) on neuroblastoma cells under glucose deprivation. The treatment aims to induce oxidative stress and activate stress-response pathways.
Take a culture of neuroblastoma cells in a multiwell plate with media.
Replace the media in wells with glucose-free neuronal media to induce oxidative stress.
Position the plate under the plasma jet nozzle at an optimal distance for uniform exposure.
Treat selected wells with a low dose of helium-based atmospheric pressure plasma jet (APPJ), which generates controlled levels of reactive oxygen and nitrogen species.
Incubate the cells to allow activation of stress-response pathways and upregulation of antioxidant enzymes in APPJ-treated wells.
Cells in untreated wells under glucose deprivation undergo further oxidative cellular damage.
Add cell viability assay reagent and incubate. Viable cells reduce the reagent to a colored product.
Measure color intensity using a microplate reader.
APPJ-treated wells exhibit increased color intensity due to higher cell viability, while untreated wells show lower intensity, confirming the neuroprotective effect of APPJ under glucose deprivation.
Adjust the distance between the nozzle of the quartz tube and the platform where the 96 well plate will be placed to three centimeters to ensure that the beam can touch the culture medium surface. Before treatment, change the culture medium in each well to RPMI 1640 without glucose medium. Now, place the plate under the atmospheric pressure plasma jet nozzle. Then treat cells in separate wells for 0, 1, 2, 4, 8, and 12 seconds.
After treatment, incubate the cells for one hour in a cell incubator at 37 degrees Celsius, following incubation at 10 microliters of Cell Counting Kit 8 solution to each well. Then, incubate the cells at 37 degrees Celsius for four hours. Finally, measure the absorbance at 450 nanometers with a microplate reader.
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