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

Jiahang Han; Zewei Wang; Yanzhao Dong; Xiaodi Zou; Haoyu Wang; Yonggang Chen; Sahar Ahmed Abdalbary; Tian Tu; Hui Lu

doi:10.3791/66097

Regulierung des Schwann-Zellwachstums durch ein gepulstes elektrisches Feld im Nanosekundenbereic...

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Neuroscience

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JoVE Journal Neuroscience

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

Regulierung des Schwann-Zellwachstums durch ein gepulstes elektrisches Feld im Nanosekundenbereich für die periphere Nervenregeneration in vitro

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09:24 min

May 3, 2024

DOI: 10.3791/66097-v

Jiahang Han*^1,2, Zewei Wang*^1,2, Yanzhao Dong*¹, Xiaodi Zou³, Haoyu Wang^1,2, Yonggang Chen¹, Sahar Ahmed Abdalbary⁴, Tian Tu⁵, Hui Lu¹

¹Department of Orthopedics, The First Affiliated Hospital, College of Medicine,Zhejiang University, ²College of Medicine,Zhejiang University, ³Department of Orthopedics,The Second Affiliated Hospital of Zhejiang Chinese Medical University, ⁴Department of Orthopedic Physical Therapy, Faculty of Physical Therapy,Nahda University in Beni Suef, ⁵Plastic & Cosmetic Center, The First Affiliated Hospital, College of Medicine,Zhejiang University

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Overview

This study presents a protocol for using nanosecond pulse electric field (nsPEF) to stimulate Schwann cells in vitro, highlighting its potential for promoting peripheral nerve regeneration. The investigation reveals that nsPEF effectively enhances the synthesis and secretion of factors related to cell behavior, indicating its efficacy for nerve regeneration methodologies.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Regenerative Medicine

Background

Peripheral nerve injury often leads to challenging regeneration processes.
Schwann cells play a crucial role in supporting nerve regeneration.
Existing methods of activating Schwann cells may have significant side effects.
This study aims to utilize nsPEF as a novel, less invasive method to stimulate these cells.

Purpose of Study

To develop an efficient method for activating Schwann cells to aid peripheral nerve regeneration.
To provide a reliable protocol for further research on nsPEF applications.
To evaluate the safety and efficiency of nsPEF in promoting cellular activity.

Methods Used

In vitro stimulation using cultured RSC96 Schwann cells.
The study involved treating cells with nsPEF at various intensities.
Key procedures included cell culture, treatment application, and various assays to measure cell proliferation and migration.
Evaluation of cellular responses was performed using CCK-8 and scratch assays.

Main Results

Cells stimulated at 5 kV/cm exhibited significantly faster proliferation and migration compared to controls.
Histological analysis showed increased S100-Beta expression in the appropriate treatment group.
Overall findings suggest that nsPEF effectively promotes Schwann cell activity, which may enhance nerve regeneration.

Conclusions

This study demonstrates that nsPEF can efficiently stimulate Schwann cells with minimal side effects.
The findings support the potential of nsPEF in developing advanced strategies for peripheral nerve repair.
Future research may build on these results to explore safe applications in vivo.

Frequently Asked Questions

What are the advantages of using nsPEF for Schwann cell stimulation?

NsPEF provides a novel method for activating Schwann cells with minimal side effects, offering a less invasive alternative compared to traditional methods.

How is the Schwann cell culture prepared for stimulation?

The RSC96 cell suspension is incubated in complete culture medium, centrifuged, and resuspended to reach the desired cell density before nsPEF treatment.

What types of outcomes can be measured following nsPEF treatment?

Outcomes include cell proliferation, migration rates, and expression of markers such as S100-Beta, which indicate cellular activity and health.

How can this method be applied in further research?

This approach can be adapted for various in vitro studies and potentially in vivo applications to explore nerve regeneration strategies.

Are there any limitations with the nsPEF method?

As the study suggests, while nsPEF shows promise, further research is needed to explore long-term effects and safety in vivo.

Hier stellen wir ein Protokoll für die Anwendung eines elektrischen Nanosekunden-Pulsfeldes (nsPEF) zur Stimulation von Schwann-Zellen in vitro vor. Die Synthese- und Sekretionsfähigkeit relevanter Faktoren und Veränderungen des Zellverhaltens bestätigten die erfolgreiche Stimulation mit nsPEF. Die Studie gibt ein positives Bild von der Methode der peripheren Nervenregeneration.

Unsere Forschung konzentriert sich auf die Förderung der peripheren Nervenregeneration durch nsPEF-stimulierte Schwanenzellen. Wir hoffen, einen neuen Weg zu entwickeln, um Schwanenzellen effizient zu aktivieren, da die Nervenregeneration in den meisten Fällen recht schwierig ist. Wir verwenden auf innovative Weise ein Nanosekunden-postelektrisches Feld nsPEF, um Schwanenzellen zu aktivieren.

Wir freuen uns, dass nsPEF ein bevorzugter Modus für die periphere Nervenregeneration mit minimalen Stimulationsnebenwirkungen sein kann. Darüber hinaus bietet unser Protokoll eine verlässliche Grundlage für die anschließende vertiefte und systematische Forschung. Unser Labor wird die Anwendung von nsPEF kontinuierlich untersuchen.

Im Hinblick auf die Nervenregeneration werden wir Tiermodelle aufbauen, um In-vivo-Tests zu fördern und die Sicherheit von nsPEF in vivo zu bewerten. Da es nur begrenzte Evidenz für die Anwendung von nsPEF bei der Regeneration von Überweisungsnerven gibt, hoffen wir, dass wir Durchbrüche erzielen können.

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Schlüsselwörter: Schwann-Zellen gepulstes elektrisches Feld im Nanosekundenbereich NsPEF periphere Nervenregeneration Zellproliferation Zellmigration neurotrophe Faktoren Myelinisierungsregulatoren GFAP