This study investigates the effects of nanosecond pulsed electric fields (nsPEF) on Schwann cells (SCs), which are crucial for peripheral nerve regeneration. The application of nsPEF disrupts SC membranes, leading to calcium influx and subsequent cellular responses that promote repair and proliferation.
Begin with Schwann cells or SCs, myelinating glial cells of the peripheral nervous system, cultured in media.
Transfer the cells to a colorimetric dish equipped with electrodes on both sides.
Place the dish on a nanosecond pulsed electric field or nsPEF device and set the desired electric field intensity.
Apply nsPEF pulses by carefully rotating the electrodes until sparks appear.
Immediately post-stimulation, separate the electrodes and continue culturing the cells.
nsPEF disrupts SC membranes, creating transient nanopores that allow calcium influx. The membranes eventually reseal.
Intracellular calcium activates calcium-binding proteins, triggering signaling pathways that upregulate genes involved in cell cycle progression.
Sustained calcium signaling also activates transcription factors that drive the dedifferentiation of SCs into a repair phenotype, followed by proliferation.
Additionally, the secretion of neurotrophic factors that support neuronal survival and growth increases.
The model is now ready to study the role of nsPEF-stimulated SCs in peripheral nerve regeneration post-injury.
To begin, resuspend the cultured RSC96 cells in 1 milliliter of DMEM culture medium, and transfer them to colorimetric dishes with electrodes on both sides. Turn on the power switch of the instrument, and rotate the knob to set the intensities of the electric field. Carefully, rotate the electrodes until sparks appear, allowing the cells to receive 5 pulses of nanosecond pulsed electric fields according to the pre-set field strength intensities. Immediately separate the two electrodes after the treatment.
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