operation of the nanosecond pulsed electric field (nspef) device to stimulate cultured rsc96 cells

nsPEF 자극 슈반(Schwann Cells)을 이용한 말초신경 재생 강화

Each year, millions of people are affected by nerve injuries involving both the peripheral nervous system (PNS) and the central nervous system (CNS)1. Studies have demonstrated that the axonal repair capacity of the CNS is quite limited after nerve injuries, while the PNS shows enhanced capacity due to the significant plasticity of SCs2. Nevertheless, achieving complete regeneration after peripheral nerve injuries remains arduous and continues to pose a significant challenge to human health3,4. Nowadays, autografts have remained a common treatment despite the drawbacks of donor site morbidity and limited availability5. This situation has prompted researchers to explore alternative therapies, including materials6, molecular factors7, and electrical stimulation (ES). As a factor promoting axonal growth and nerve regeneration8, choosing an appropriate method of ES and exploring the relationship between ES and SCs become essential.
SCs are the main glial cells of the PNS, playing a crucial role in the regeneration of the PNS9,10. Following peripheral nerve injuries, SCs undergo rapid activation, extensive reprogramming2,&#160;and transition from a myelin-forming state to a growth-supportive morphology to conduct the regeneration of the nerve2. A substantial proliferation of SCs occurs at the distal end of the injured nerve, while SCs of the distal stump undergo proliferation and elongation to form Bungner&#39;s band, which are necessary to guide axons to grow towards the target organ11. Moreover, SCs from the proximal and distal nerve stumps migrate into the nerve bridge to form SC cords promoting axon regeneration12. Furthermore, previous studies have demonstrated that the synthesis and secretion of relevant factors related to SCs change in cases of peripheral nerve regeneration, including transcriptional factors13, neurotrophic factors14, and myelination regulators13. This also provides indicators for assessing the activity of SCs. Based on these, the promotion of SC proliferation, migration, synthesis, and secretion of relevant factors have been extensively investigated for improving peripheral nerve regeneration15.
Previous studies have demonstrated the possibility of using ES for nerve regeneration1. A widely accepted explanation is that ES can induce depolarization of cell membranes, alter membrane potential, and affect membrane protein functions by changing the charge distributions on these biomolecules1. However, widely applied Intense PEF may cause severe pain, involuntary muscle contractions, and heart fibrillation8. It also increases creatine kinase (CK) activity, decreases muscle strength, and induces the development of delayed onset muscle soreness (DOMS)16. nsPEF is an emerging technique that stimulates test subjects with high-voltage electric fields within a nanosecond pulse duration, and it is gradually being used in cellular-level research17,18. Previous studies have reported that the possible rationale of nsPEF promoting cell proliferation and organelle activity is the formation of membrane nanopores and the activation of ionic channels, which leads to an increase in cytoplasmic Ca2+ concentration19. nsPEF utilizes pulse power technology to charge the cell membrane, producing pulses characterized by short duration, rapid rise time, high power, and low energy density20. These characteristics suggest that nsPEF may be a preferred mode with minimal stimulation side effects8. Furthermore, nsPEF offers advantages such as minimally invasive procedures, reversibility, adjustability, and non-destructiveness to neural tissues compared to surgical interventions. One mainstream research direction of nsPEF in the biomedical field is its application for tumor tissue ablation using high-energy electric field stimulation21,22,23. Some research results indicate that 12-nsPEF can stimulate peripheral nerves without causing damage24. However, at present, there is limited evidence regarding the application of nsPEF in the field of nerve regeneration. Moreover, stimulating SCs using nsPEF is a pioneering attempt, contributing to further in vivo and clinical research. This study explores whether nsPEF stimulation of SCs can promote nerve regeneration and provide a reliable basis for subsequent in-depth and systematic research.

저자 스포트라이트: 말초 신경 재생을 촉진하기 위한 nsPEF의 혁신적인 사용

Regulating Schwann Cell Growth by Nanosecond Pulsed Electric Field for Peripheral Nerve Regeneration In Vitro(체외 말초 신경 재생을 위한 나노초 펄스 전기장에 의한 Schwann 세포 성장 조절)

Our research focuses on promoting peripheral nerve regeneration by nsPEF stimulated swan cells. We hope to develop a new way to activate swan cells efficiently as nerve regeneration is quite difficult in most cases. We innovatively use a nanosecond post electric field nsPEF to activate swan cells.We are delighted to find that nsPEF may be a preferred mode for peripheral nerve regeneration with minimal stimulation side effects. Besides our protocol provide a reliable basis for subsequent in depth and systematic research. Our laboratory will continually explore the application of nsPEF.In terms of nerve regeneration, we will build up animal models to promote in vivo testing and evaluate the safety of nsPEF in vivo. As there is limited evidence regarding the application of nsPEF in referral nerve regeneration, we hope we can make breakthroughs.

배양된 RSC96 세포를 자극하기 위한 나노초 펄스 전기장(nsPEF) 장치의 작동

먼저 배양된 RSC96 세포를 1mL의 DMEM 배양 배지에 재현탁시키고 양면에 전극이 있는 비색 접시로 옮깁니다. 기기의 전원 스위치를 켜고 손잡이를 돌려 전기장의 강도를 설정합니다. 스파크가 나타날 때까지 전극을 조심스럽게 회전시켜 셀이 사전 설정된 전계 강도 강도에 따라 나노초 펄스 전기장의 5개 펄스를 받을 수 있도록 합니다.치료 후 즉시 두 전극을 분리하십시오.

operation-nanosecond-pulsed-electric-field-nspef-device-to-stimulate

Research

JoVE Journal

Neuroscience

Operation of the Nanosecond Pulsed Electric Field (nsPEF) Device to Stimulate Cultured RSC96 Cells

44 조회수.  Zhejiang University. 먼저 배양된 RSC96 세포를 1mL의 DMEM 배양 배지에 재현탁시키고 양면에 전극이 있는 비색 접시로 옮깁니다. 기기의 전원 스위치를 켜고 손잡이를 돌려 전기장의 강도를 설정합니다. 스파크가 나타날 때까지 전극을 조심스럽게 회전시켜 셀이 사전 설정된 전계 강도 강도에 따라 나노초 펄스 전기장의 5개 펄스를 받을 수 있도록 합니다.치료 후 즉시 두 전극을 분리하십시오....