Recovering the regenerative potential in chronically injured nerves by using conditioning electrical stimulation

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  • 1 Division of Plastic Surgery, Department of Surgery, Faculty of Medicine and Dentistry, University of Alberta;
  • | 2 Division of Anatomy, Department of Surgery, Faculty of Medicine and Dentistry, University of Alberta;
  • | 3 Division of Physical Medicine and Rehabilitation, Department of Medicine, Faculty of Medicine and Dentistry, University of Alberta; and
  • | 4 Department of Anthropology, Faculty of Science, University of Alberta, Edmonton, Alberta, Canada
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OBJECTIVE

Chronically injured nerves pose a significant clinical challenge despite surgical management. There is no clinically feasible perioperative technique to upregulate a proregenerative environment in a chronic nerve injury. Conditioning electrical stimulation (CES) significantly improves sensorimotor recovery following acute nerve injury to the tibial and common fibular nerves. The authors’ objective was to determine if CES could foster a proregenerative environment following chronically injured nerve reconstruction.

METHODS

The tibial nerve of 60 Sprague Dawley rats was cut, and the proximal ends were inserted into the hamstring muscles to prevent spontaneous reinnervation. Eleven weeks postinjury, these chronically injured animals were randomized, and half were treated with CES proximal to the tibial nerve cut site. Three days later, 24 animals were killed to evaluate the effects of CES on the expression of regeneration-associated genes at the cell body (n = 18) and Schwann cell proliferation (n = 6). In the remaining animals, the tibial nerve defect was reconstructed using a 10-mm isograft. Length of nerve regeneration was assessed 3 weeks postgrafting (n = 16), and functional recovery was evaluated weekly between 7 and 19 weeks of regeneration (n = 20).

RESULTS

Three weeks after nerve isograft surgery, tibial nerves treated with CES prior to grafting had a significantly longer length of nerve regeneration (p < 0.01). Von Frey analysis identified improved sensory recovery among animals treated with CES (p < 0.01). Motor reinnervation, assessed by kinetics, kinematics, and skilled motor tasks, showed significant recovery (p < 0.05 to p < 0.001). These findings were supported by immunohistochemical quantification of motor endplate reinnervation (p < 0.05). Mechanisms to support the role of CES in reinvigorating the regenerative response were assessed, and it was demonstrated that CES increased the proliferation of Schwann cells in chronically injured nerves (p < 0.05). Furthermore, CES upregulated regeneration-associated gene expression to increase growth-associated protein–43 (GAP-43), phosphorylated cAMP response element binding protein (pCREB) at the neuronal cell bodies, and upregulated glial fibrillary acidic protein expression in the surrounding satellite glial cells (p < 0.05 to p < 0.001).

CONCLUSIONS

Regeneration following chronic axotomy is impaired due to downregulation of the proregenerative environment generated following nerve injury. CES delivered to a chronically injured nerve influences the cell body and the nerve to re-upregulate an environment that accelerates axon regeneration, resulting in significant improvements in sensory and motor functional recovery. Percutaneous CES may be a preoperative strategy to significantly improve outcomes for patients undergoing delayed nerve reconstruction.

ABBREVIATIONS

ATF3 = activation transcription factor 3; CES = conditioning electrical stimulation; DRG = dorsal root ganglion; GAP-43 = growth-associated protein–43; GFAP = glial fibrillary acidic protein; NF200 = neurofilament-200; NMJ = neuromuscular junction; PBS = phosphate-buffered saline; pCREB = phosphorylated cAMP response element binding protein; RAG = regeneration-associated gene.

Illustration from Serrato-Avila (pp 1410–1423). Copyright Johns Hopkins University, Art as Applied to Medicine. Published with permission.

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  • 1

    Gordon T, Tetzlaff W. Regeneration-associated genes decline in chronically injured rat sciatic motoneurons. Eur J Neurosci. 2015;42(10):27832791.

  • 2

    Fu SY, Gordon T. Contributing factors to poor functional recovery after delayed nerve repair: prolonged axotomy. J Neurosci. 1995;15(5 Pt 2):38763885.

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 3

    Sulaiman OAR, Gordon T. Role of chronic Schwann cell denervation in poor functional recovery after nerve injuries and experimental strategies to combat it. Neurosurgery. 2009;65(4 Suppl):A105A114.

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 4

    Sulaiman OAR, Gordon T. Effects of short- and long-term Schwann cell denervation on peripheral nerve regeneration, myelination, and size. Glia. 2000;32(3):234246.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 5

    Senger JB, Verge VMK, Chan KM, Webber CA. The nerve conditioning lesion: a strategy to enhance nerve regeneration. Ann Neurol. 2018;83(4):691702.

  • 6

    Senger JLB, Verge VMK, Macandili HSJ, et al. Electrical stimulation as a conditioning strategy for promoting and accelerating peripheral nerve regeneration. Exp Neurol. 2018;302:7584.

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 7

    Senger JL, Chan KM, Macandili H, et al. Conditioning electrical stimulation promotes functional nerve regeneration. Exp Neurol. 2019;315:6071.

  • 8

    Senger JB, Chan KM, Webber CA. Conditioning electrical stimulation is superior to postoperative electrical stimulation, resulting in enhanced nerve regeneration and functional recovery. Exp Neurol. 2020;325:113147.

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 9

    Senger JB, Chan AWM, Chan KM, et al. Conditioning electrical stimulation is superior to postoperative electrical stimulation in enhanced regeneration and functional recovery following nerve graft repair. Neurorehabil Neural Repair. 2020;34(4):299308.

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 10

    Senger JB, Rabey KN, Morhart MJ, et al. Conditioning electrical stimulation accelerates regeneration in nerve transfers. Ann Neurol. 2020;88(2):363374.

  • 11

    Al-Majed AA, Neumann CM, Brushart TM, Gordon T. Brief electrical stimulation promotes the speed and accuracy of motor axonal regeneration. J Neurosci. 2000;20(7):26022608.

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 12

    Andersen PL, Webber CA, Kimura KA, Schreyer DJ. Cyclic AMP prevents an increase in GAP-43 but promotes neurite growth in cultured adult rat dorsal root ganglion neurons. Exp Neurol. 2000;166(1):153165.

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 13

    Cheng C, Webber CA, Wang J, et al. Activated RHOA and peripheral axon regeneration. Exp Neurol. 2008;212(2):358369.

  • 14

    Webber CA, Zochodne DW. Preparation of adult rat sensory neuron cultures and their application to growth cone turning assays. Methods Mol Biol. 2018;1727:8192.

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 15

    Metz GA, Whishaw IQ. The ladder rung walking task: a scoring system and its practical application. J Vis Exp. 2009;(28):1204.

  • 16

    Charles JP, Cappellari O, Hutchinson JR. A dynamic simulation of musculoskeletal function in the mouse hindlimb during trotting locomotion. Front Bioeng Biotechnol. 2018;6:61.

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 17

    Kemp SWP, Alant J, Walsh SK, et al. Behavioural and anatomical analysis of selective tibial nerve branch transfer to the deep peroneal nerve in the rat. Eur J Neurosci. 2010;31(6):10741090.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 18

    Eftaxiopoulou T, Macdonald W, Britzman D, Bull AMJ. Gait compensations in rats after a temporary nerve palsy quantified using temporo-spatial and kinematic parameters. J Neurosci Methods. 2014;232:1623.

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 19

    McNeill JN, Wu CL, Rabey KN, et al. Life-long caloric restriction does not alter the severity of age-related osteoarthritis. Age (Dordr). 2014;36(4):9669.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 20

    Allen KD, Mata BA, Gabr MA, et al. Kinematic and dynamic gait compensations resulting from knee instability in a rat model of osteoarthritis. Arthritis Res Ther. 2012;14(2):R78.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 21

    Gordon T, Amirjani N, Edwards DC, Chan KM. Brief post-surgical electrical stimulation accelerates axon regeneration and muscle reinnervation without affecting the functional measures in carpal tunnel syndrome patients. Exp Neurol. 2010;223(1):192202.

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 22

    Soilu-Hänninen M, Ekert P, Bucci T, et al. Nerve growth factor signaling through p75 induces apoptosis in Schwann cells via a Bcl-2-independent pathway. J Neurosci. 1999;19(12):48284838.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 23

    Gordon T, Sulaiman O, Boyd JG. Experimental strategies to promote functional recovery after peripheral nerve injuries. J Peripher Nerv Syst. 2003;8(4):236250.

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 24

    Gordon T, Wood P, Sulaiman OAR. Long-term denervated rat Schwann cells retain their capacity to proliferate and to myelinate axons in vitro. Front Cell Neurosci. 2019;12:511.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 25

    Dienes JA, Hu X, Janson KD, et al. Analysis and modeling of rat gait biomechanical deficits in response to volumetric muscle loss injury. Front Bioeng Biotechnol. 2019;7:146.

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 26

    Tajino J, Ito A, Tanima M, et al. Three-dimensional motion analysis for comprehensive understanding of gait characteristics after sciatic nerve lesion in rodents. Sci Rep. 2018;8(1):13585.

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 27

    Howard CS, Blakeney DC, Medige J, et al. Functional assessment in the rat by ground reaction forces. J Biomech. 2000;33(6):751757.

  • 28

    Varejão ASP, Cabrita AM, Meek MF, et al. Ankle kinematics to evaluate functional recovery in crushed rat sciatic nerve. Muscle Nerve. 2003;27(6):706714.

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