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

שיפור התחדשות עצבית היקפית עם תאי Schwann מעוררי nsPEF

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.

מחבר זרקור: שימוש חדשני ב- nsPEF להגברת התחדשות העצבים ההיקפיים

ויסות צמיחת תאי Schwann על ידי שדה חשמלי פועם ננו-שנייה להתחדשות עצבית היקפית במבחנה

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.

הפעלת התקן שדה חשמלי פועם ננו-שנייה (nsPEF) לגירוי תאי RSC96 בתרבית

כדי להתחיל, להשעות מחדש את תאי RSC96 בתרבית במיליליטר אחד של מדיום תרבית DMEM, ולהעביר אותם צלחות colorimetric עם אלקטרודות משני הצדדים. הפעל את מתג ההפעלה של המכשיר וסובב את הידית כדי להגדיר את עוצמות השדה החשמלי. סובבו בזהירות את האלקטרודות עד להופעת ניצוצות, מה שמאפשר לתאים לקבל חמישה פולסים של שדות חשמליים פועמים בננו-שנייה בהתאם לעוצמות חוזק השדה שנקבעו מראש.יש להפריד מיד בין שתי האלקטרודות לאחר הטיפול.

Research

JoVE Journal

Neuroscience

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

Regulating Schwann Cell Growth by Nanosecond Pulsed Electric Field for Peripheral Nerve Regeneration In Vitro

Schwann cells (SCs) are myelinating cells of the peripheral nervous system, playing a crucial role in peripheral nerve regeneration. Nanosecond Pulse ...

Preparing Cryopreserved RSC96 Cells for Nanosecond Pulsed Electric Field Stimulation

הכנת תאי RSC96 שמורים בהקפאה לגירוי שדה חשמלי פועם ננו-שנייה

Validating the Cellular Proliferation and Migration of Nanosecond Pulsed Electric Field (nsPEF) Stimulated RSC96 Cells

אימות ההתפשטות התא והנדידה של ננו-שניות של תאי RSC96 המעוררים את תאי RSC96

Detecting the Expression of the Neurotrophic and Transcription Factors in Nanosecond Pulsed Electric Field (nsPEF) Stimulated RSC96 Cells

זיהוי הביטוי של גורמי הנוירוטרופיה והשעתוק בתאי RSC96 מעוררי שדה חשמלי פועם ננו-שניות (nsPEF)