Brief electrical stimulation improves nerve regeneration after delayed repair in Sprague Dawley rats

Functional recovery after peripheral nerve injury and surgical repair declines with time and distance because the injured neurons without target contacts (chronic axotomy) progressively lose their regenerative capacity and chronically denervated Schwann cells (SCs) atrophy and fail to support axon regeneration. Findings that brief low frequency electrical stimulation (ES) accelerates axon outgrowth and muscle reinnervation after immediate nerve surgery in rats and human patients suggest that ES might improve regeneration after delayed nerve repair. To test this hypothesis, common peroneal (CP) neurons were chronically axotomized and/or tibial (TIB) SCs and ankle extensor muscles were chronically denervated by transection and ligation in rats. The CP and TIB nerves were cross-sutured after three months and subjected to either sham or one hour 20Hz ES. Using retrograde tracing, we found that ES significantly increased the numbers of both motor and sensory neurons that regenerated their axons after a three month period of chronic CP axotomy and/or chronic TIB SC denervation. Muscle and motor unit forces recorded to determine the numbers of neurons that reinnervated gastrocnemius muscle demonstrated that ES significantly increased the numbers of motoneurons that reinnervated chronically denervated muscles. We conclude that electrical stimulation of chronically axotomized motor and sensory neurons is effective in accelerating axon outgrowth into chronically denervated nerve stumps and improving target reinnervation after delayed nerve repair. Possible mechanisms for the efficacy of ES in promoting axon regeneration and target reinnervation after delayed nerve repair include the upregulation of neurotrophic factors.

[1]  M. Raji,et al.  The Basis for Diminished Functional Recovery after Delayed Peripheral Nerve Repair , 2011, The Journal of Neuroscience.

[2]  W. Tetzlaff,et al.  Rapid induction of the major embryonic alpha-tubulin mRNA, T alpha 1, during nerve regeneration in adult rats , 1989, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[3]  Charles Rohde,et al.  Electrical stimulation restores the specificity of sensory axon regeneration , 2005, Experimental Neurology.

[4]  A. Höke,et al.  Advances in peripheral nerve regeneration , 2013, Nature Reviews Neurology.

[5]  T. Gordon,et al.  BDNF/TrkB signaling regulates HNK-1 carbohydrate expression in regenerating motor nerves and promotes functional recovery after peripheral nerve repair , 2006, Experimental Neurology.

[6]  R. Redett,et al.  Schwann Cells Express Motor and Sensory Phenotypes That Regulate Axon Regeneration , 2006, The Journal of Neuroscience.

[7]  A. Koppes,et al.  Neurite outgrowth on electrospun PLLA fibers is enhanced by exogenous electrical stimulation , 2014, Journal of neural engineering.

[8]  A. English,et al.  Cooperative Roles of BDNF Expression in Neurons and Schwann Cells Are Modulated by Exercise to Facilitate Nerve Regeneration , 2012, The Journal of Neuroscience.

[9]  U. Rutishauser,et al.  Intrinsic neuronal properties control selective targeting of regenerating motoneurons. , 2008, Brain : a journal of neurology.

[10]  Tessa Gordon,et al.  Brief post-surgical electrical stimulation accelerates axon regeneration and muscle reinnervation without affecting the functional measures in carpal tunnel syndrome patients , 2010, Experimental Neurology.

[11]  T Gordon,et al.  Brief Electrical Stimulation Promotes the Speed and Accuracy of Motor Axonal Regeneration , 2000, The Journal of Neuroscience.

[12]  Tessa Gordon,et al.  Electrical Stimulation Promotes Motoneuron Regeneration without Increasing Its Speed or Conditioning the Neuron , 2002, The Journal of Neuroscience.

[13]  T. Gordon,et al.  Time course of preferential motor unit loss in the SOD1G93A mouse model of amyotrophic lateral sclerosis , 2007, Neurobiology of Disease.

[14]  Ryan G. Porter,et al.  Effects of electrical stimulation and gonadal steroids on rat facial nerve regenerative properties. , 2009, Restorative neurology and neuroscience.

[15]  T. Gordon,et al.  A Decline in Glial Cell-Line-Derived Neurotrophic Factor Expression Is Associated with Impaired Regeneration after Long-Term Schwann Cell Denervation , 2002, Experimental Neurology.

[16]  T Gordon,et al.  Contributing factors to poor functional recovery after delayed nerve repair: prolonged denervation , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[17]  T. Gordon,et al.  Accelerated axon outgrowth, guidance, and target reinnervation across nerve transection gaps following a brief electrical stimulation paradigm. , 2012, Journal of neurosurgery.

[18]  W. Tetzlaff,et al.  Changes in cytoskeletal protein synthesis following axon injury and during axon regeneration , 2007, Molecular Neurobiology.

[19]  Kathryn J. Jones,et al.  Electrical stimulation and testosterone differentially enhance expression of regeneration-associated genes , 2010, Experimental Neurology.

[20]  Zhuojing Luo,et al.  Electrical stimulation accelerates nerve regeneration and functional recovery in delayed peripheral nerve injury in rats , 2013, The European journal of neuroscience.

[21]  T. Gordon,et al.  Electrical stimulation accelerates and increases expression of BDNF and trkB mRNA in regenerating rat femoral motoneurons. , 2000, The European journal of neuroscience.

[22]  Xavier Navarro,et al.  Effects of activity-dependent strategies on regeneration and plasticity after peripheral nerve injuries. , 2011, Annals of anatomy = Anatomischer Anzeiger : official organ of the Anatomische Gesellschaft.

[23]  X. Navarro,et al.  Passive and active exercise improve regeneration and muscle reinnervation after peripheral nerve injury in the rat , 2011, Muscle & nerve.

[24]  W. Nix,et al.  Electrical stimulation of regenerating nerve and its effect on motor recovery , 1983, Brain Research.

[25]  T. Gordon,et al.  Neurotrophic factors and their receptors in axonal regeneration and functional recovery after peripheral nerve injury , 2003, Molecular Neurobiology.

[26]  A. English,et al.  Effects of treadmill training on functional recovery following peripheral nerve injury in rats. , 2013, Journal of neurophysiology.

[27]  W. Ding,et al.  RETRACTED: Electrical stimulation enhanced remyelination of injured sciatic nerves by increasing neurotrophins , 2010, Neuroscience.

[28]  T. Gordon,et al.  ROLE OF CHRONIC SCHWANN CELL DENERVATION IN POOR FUNCTIONAL RECOVERY AFTER NERVE INJURIES AND EXPERIMENTAL STRATEGIES TO COMBAT IT , 2009, Neurosurgery.

[29]  U. Rutishauser,et al.  Polysialylated Neural Cell Adhesion Molecule Is Necessary for Selective Targeting of Regenerating Motor Neurons , 2005, The Journal of Neuroscience.

[30]  T Gordon,et al.  Increased Neuromuscular Activity Reduces Sprouting in Partially Denervated Muscles , 2001, The Journal of Neuroscience.

[31]  T. Gordon,et al.  The expression of the low affinity nerve growth factor receptor in long‐term denervated Schwann cells , 1997, Glia.

[32]  T. Gordon,et al.  Experimental and clinical evidence for use of decellularized nerve allografts in peripheral nerve gap reconstruction. , 2013, Tissue engineering. Part B, Reviews.

[33]  T. Gordon,et al.  PROLONGED TARGET DEPRIVATION REDUCES THE CAPACITY OF INJURED MOTONEURONS TO REGENERATE , 2007, Neurosurgery.

[34]  W. E. Watson.  Some metabolic responses of axotomized neurones to contact between their axons and denervated muscle , 1970, The Journal of physiology.

[35]  J. Jahng,et al.  Thirty minutes of low intensity electrical stimulation promotes nerve regeneration after sciatic nerve crush injury in a rat model. , 2010, Acta neurologica Belgica.

[36]  R. Redett,et al.  Schwann cell phenotype is regulated by axon modality and central–peripheral location, and persists in vitro , 2013, Experimental Neurology.

[37]  S. Marzo,et al.  Combinatorial treatments enhance recovery following facial nerve crush , 2010, The Laryngoscope.

[38]  R. Stein,et al.  Time course and extent of recovery in reinnervated motor units of cat triceps surae muscles , 1982, The Journal of physiology.

[39]  X. Navarro,et al.  Electrical stimulation combined with exercise increase axonal regeneration after peripheral nerve injury , 2009, Experimental Neurology.

[40]  G. Kreutzberg,et al.  Changes in cytoskeletal proteins in the rat facial nucleus following axotomy , 1988, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[41]  A. Hudson,et al.  Timing of peripheral nerve repair: important local neuropathological factors. , 1977, Clinical neurosurgery.

[42]  K. Fouad,et al.  Electrical stimulation of intact peripheral sensory axons in rats promotes outgrowth of their central projections , 2008, Experimental Neurology.

[43]  Lora A Major,et al.  Method for counting motor units in mice and validation using a mathematical model. , 2007, Journal of neurophysiology.

[44]  S. Marzo,et al.  Accelerating functional recovery after rat facial nerve injury: Effects of gonadal steroids and electrical stimulation , 2008, Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery.

[45]  T. Gordon,et al.  Effects of short‐ and long‐term Schwann cell denervation on peripheral nerve regeneration, myelination, and size , 2000, Glia.

[46]  Chun-Hsu Yao,et al.  Timing of Applying Electrical Stimulation Is an Important Factor Deciding the Success Rate and Maturity of Regenerating Rat Sciatic Nerves , 2010, Neurorehabilitation and neural repair.

[47]  A. Koppes,et al.  Neurite outgrowth is significantly increased by the simultaneous presentation of Schwann cells and moderate exogenous electric fields , 2011, Journal of neural engineering.

[48]  Tessa Gordon,et al.  Electrical stimulation promotes sensory neuron regeneration and growth-associated gene expression , 2007, Experimental Neurology.

[49]  T Gordon,et al.  Contributing factors to poor functional recovery after delayed nerve repair: prolonged axotomy , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[50]  T. Gordon,et al.  Reduced expression of regeneration associated genes in chronically axotomized facial motoneurons , 2015, Experimental Neurology.

[51]  S. Marzo,et al.  Electrical stimulation facilitates rat facial nerve recovery from a crush injury , 2008, Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery.

[52]  A. Höke,et al.  Pleiotrophin is a neurotrophic factor for spinal motor neurons , 2007, Proceedings of the National Academy of Sciences.

[53]  A. English,et al.  Sex differences in the effectiveness of treadmill training in enhancing axon regeneration in injured peripheral nerves , 2012, Developmental neurobiology.

[54]  T. Gordon,et al.  Rolipram-induced elevation of cAMP or chondroitinase ABC breakdown of inhibitory proteoglycans in the extracellular matrix promotes peripheral nerve regeneration , 2010, Experimental Neurology.

[55]  Zhuojing Luo,et al.  Electrical stimulation induces calcium‐dependent release of NGF from cultured Schwann cells , 2009, Glia.

[56]  A. Irintchev,et al.  One hour electrical stimulation accelerates functional recovery after femoral nerve repair , 2007, Experimental Neurology.

[57]  Tessa Gordon,et al.  Electrical Stimulation Accelerates and Enhances Expression of Regeneration-Associated Genes in Regenerating Rat Femoral Motoneurons , 2004, Cellular and Molecular Neurobiology.

[58]  M. Abercrombie.  Estimation of nuclear population from microtome sections , 1946, The Anatomical record.

[59]  Tessa Gordon,et al.  Brief electrical stimulation accelerates axon regeneration in the peripheral nervous system and promotes sensory axon regeneration in the central nervous system. , 2009, Motor control.

[60]  Göran Lundborg,et al.  Nerve Injury and Repair , 1988 .

[61]  T. Gordon,et al.  A dose‐dependent facilitation and inhibition of peripheral nerve regeneration by brain‐derived neurotrophic factor , 2002, The European journal of neuroscience.

[62]  A. English,et al.  Chondroitinase ABC reduces time to muscle reinnervation and improves functional recovery after sciatic nerve transection in rats. , 2012, Journal of neurophysiology.

[63]  D. Sengelaub,et al.  Enhancement of peripheral nerve regeneration due to treadmill training and electrical stimulation is dependent on androgen receptor signaling , 2014, Developmental neurobiology.

[64]  T. Gordon,et al.  cAMP promotes neurite outgrowth and extension through protein kinase A but independently of Erk activation in cultured rat motoneurons , 2008, Neuropharmacology.

[65]  C. Rohde,et al.  Pathway sampling by regenerating peripheral axons , 2005, The Journal of comparative neurology.

[66]  K. Fargo,et al.  Single session of brief electrical stimulation immediately following crush injury enhances functional recovery of rat facial nerve. , 2012, Journal of rehabilitation research and development.

[67]  Zhuojing Luo,et al.  Electrical Stimulation Accelerates Motor Functional Recovery in Autograft-Repaired 10 mm Femoral Nerve Gap in Rats , 2009 .

[68]  Jia-Horng Lin,et al.  Effects of Electrical Stimulation at Different Frequencies on Regeneration of Transected Peripheral Nerve , 2008, Neurorehabilitation and neural repair.

[69]  D. Thompson,et al.  Electrical stimulation of schwann cells promotes sustained increases in neurite outgrowth. , 2013, Tissue engineering. Part A.

[70]  A. English,et al.  Electrical stimulation promotes peripheral axon regeneration by enhanced neuronal neurotrophin signaling , 2007, Developmental neurobiology.

[71]  T. Gordon,et al.  Neurobiology of peripheral nerve injury, regeneration, and functional recovery: from bench top research to bedside application. , 2013, The Ochsner journal.