Protective Effect of Moderate Exogenous Electric Field Stimulation on Activating Netrin-1/DCC Expression Against Mechanical Stretch-Induced Injury in Spinal Cord Neurons

  • Meili Liu
  • Chuanwei Yin
  • Zhengtai Jia
  • Kun Li
  • Zhifa Zhang
  • Yuchen Zhao
  • Xianghui Gong
  • Xiaoyu Liu
  • Ping Li
  • Yubo Fan
Original Article
  • 36 Downloads

Abstract

Nerve cells detect and respond to electric field stimulation and extrinsic chemical guidance cues during development and regeneration; therefore, the development and optimization of an approach for functional neuronal regeneration are necessary for a nerve injury. In this study, we proposed using electric field stimulation to repair a nerve injury triggered by serious mechanical stretch loading. A device that provides continuous mechanical stretch and constant electric field stimulation was designed. Primary dissociated spinal cord neurons were stimulated by mechanical stretch (tensile strain 2.5–10%) at different times (1, 4, 8, and 12 h) to set up a moderate nerve injury model. Stimulated samples were evaluated with respect to cell viability, density, and axonal elongation by the MTT and immunofluorescence assays. The results indicated that mechanical stretch (S, 5% tensile strain, 4 h) caused moderate axonal injury, resulting in significant loss of cell viability and a decrease in cell density. However, injured spinal cord neurons became viable after electric field stimulation (E, 33 mA/m2, 4 h) in the fluorescein diacetate assay. In addition, neuronal viability, density, and elongation increased significantly after electric field stimulation compared with those of stretch-injured neurons. Moreover, electric field stimulation significantly activated the axonal guidance cues Netrin-1 and deleted in colorectal cancer (DCC) receptor expression compared with the stretch-injury group. These results indicate that electric stimulation activates synergistic guidance cues of expression to improve axonal growth relevant to nerve injuries. Our study provides new insight into neuronal regeneration.

Keywords

Electric field Mechanical stretch Axonal guidance growth Netrin-1 DCC Spinal cord neurons 

Notes

Acknowledgments

We thank Michael Anderson for our English editing.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants performed by any of the authors.

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Informed Consent

Informed consent was obtained from all individual participants included in the study.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Meili Liu
    • 1
    • 2
  • Chuanwei Yin
    • 1
    • 2
  • Zhengtai Jia
    • 1
    • 2
  • Kun Li
    • 1
    • 2
  • Zhifa Zhang
    • 1
    • 2
  • Yuchen Zhao
    • 1
    • 2
  • Xianghui Gong
    • 1
    • 2
  • Xiaoyu Liu
    • 1
    • 2
  • Ping Li
    • 1
    • 2
  • Yubo Fan
    • 1
    • 2
    • 3
  1. 1.Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Biological Science and Medical EngineeringBeihang UniversityBeijingChina
  2. 2.Beijing Advanced Innovation Centre for Biomedical EngineeringBeihang UniversityBeijingChina
  3. 3.Beijing Key Laboratory of Rehabilitation Technical Aids for Old-Age DisabilityNational Research Center for Rehabilitation Technical AidsBeijingChina

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