J Reconstr Microsurg 2016; 32(06): 491-497
DOI: 10.1055/s-0036-1579540
Original Article
Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

Electrical Nerve Stimulation Enhances Perilesional Branching after Nerve Grafting but Fails to Increase Regeneration Speed in a Murine Model

Christian Witzel
1   Plastic and Reconstructive Surgery, Charité-Universitätsmedizin Berlin, Germany
,
Thomas M. Brushart
2   Department of Orthopaedic Surgery, Johns Hopkins Medical Institutions, Baltimore, Maryland
,
Georgios Koulaxouzidis*
3   Department of Plastic and Hand Surgery, University of Freiburg Medical Centre, Freiburg, Germany
,
Manfred Infanger*
4   Department of Plastic, Aesthetic and Hand Surgery, Otto von Guericke University, Magdeburg, Germany
› Author Affiliations
Further Information

Publication History

23 June 2015

09 January 2016

Publication Date:
14 March 2016 (online)

Abstract

Background Electrical stimulation immediately following nerve lesion helps regenerating axons cross the subsequently grafted nerve repair site. However, the results and the mechanisms remain open to debate. Some findings show that stimulation after crush injury increases axonal crossing of the repair site without affecting regeneration speed. Others show that stimulation after transection and fibrin glue repair doubles regeneration distance.

Methods Using a sciatic-nerve-transection-graft in vivo model, we investigated the morphological behavior of regenerating axons around the repair site after unilateral nerve stimulation (20 Hz, 1 hour). With mice expressing axonal fluorescent proteins (thy1-YFP), we were able to calculate the following at 5 and 7 days: percentage of regenerating axons and arborizing axons, branches per axon, and regeneration distance and speed.

Results Brief stimulation significantly increases the percentage of regenerating axons (5 days: 35.5 vs. 27.3% nonstimulated, p < 0.05; 7 days: 43.3 vs. 33.9% nonstimulated, p < 0.05), mainly by increasing arborizing axons (5 days: 49.3 [4.4] vs. 33.9 [4.1]% [p < 0.001]; 7 days: 42.2 [5.6] vs. 33.2 [3.1]% [p < 0.001]). Neither branches per arborizing axon nor regeneration speed were affected.

Conclusion Our morphological data analysis revealed that electrical stimulation in this model increases axonal crossing of the repair site and promotes homogeneous perilesional branching, but does not affect regeneration speed.

* Both the authors contributed equally to the study.


 
  • References

  • 1 Giannessi E, Coli A, Stornelli MR , et al. An autologously generated platelet-rich plasma suturable membrane may enhance peripheral nerve regeneration after neurorraphy in an acute injury model of sciatic nerve neurotmesis. J Reconstr Microsurg 2014; 30 (9) 617-626
  • 2 Fowler JR, Lavasani M, Huard J, Goitz RJ. Biologic strategies to improve nerve regeneration after peripheral nerve repair. J Reconstr Microsurg 2015; 31 (4) 243-248
  • 3 Wang P, Zhao J, Jiang B, Zhang Y. Use of small gap anastomosis for the repair of peripheral nerve injury by cutting and sleeve jointing the epineurium. J Reconstr Microsurg 2015; 31 (4) 268-276
  • 4 Evangelista MS, Perez M, Salibian AA , et al. Single-lumen and multi-lumen poly(ethylene glycol) nerve conduits fabricated by stereolithography for peripheral nerve regeneration in vivo. J Reconstr Microsurg 2015; 31 (5) 327-335
  • 5 Rinker BD, Ingari JV, Greenberg JA, Thayer WP, Safa B, Buncke GM. Outcomes of short-gap sensory nerve injuries reconstructed with processed nerve allografts from a multicenter registry study. J Reconstr Microsurg 2015; 31 (5) 384-390
  • 6 Brushart TM, Hoffman PN, Royall RM, Murinson BB, Witzel C, Gordon T. Electrical stimulation promotes motoneuron regeneration without increasing its speed or conditioning the neuron. J Neurosci 2002; 22 (15) 6631-6638
  • 7 Fawcett JW. Intrinsic neuronal determinants of regeneration. Trends Neurosci 1992; 15 (1) 5-8
  • 8 Al-Majed AA, Brushart TM, Gordon T. Electrical stimulation accelerates and increases expression of BDNF and trkB mRNA in regenerating rat femoral motoneurons. Eur J Neurosci 2000; 12 (12) 4381-4390
  • 9 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) 2602-2608
  • 10 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) 192-202
  • 11 Gordon T, Chan KM, Sulaiman OA, Udina E, Amirjani N, Brushart TM. Accelerating axon growth to overcome limitations in functional recovery after peripheral nerve injury. Neurosurgery 2009; 65 (4, Suppl) A132-A144
  • 12 English AW, Schwartz G, Meador W, Sabatier MJ, Mulligan A. Electrical stimulation promotes peripheral axon regeneration by enhanced neuronal neurotrophin signaling. Dev Neurobiol 2007; 67 (2) 158-172
  • 13 English AW, Meador W, Carrasco DI. Neurotrophin-4/5 is required for the early growth of regenerating axons in peripheral nerves. Eur J Neurosci 2005; 21 (10) 2624-2634
  • 14 Witzel C, Rohde C, Brushart TM. Pathway sampling by regenerating peripheral axons. J Comp Neurol 2005; 485 (3) 183-190
  • 15 Koulaxouzidis G, Reim G, Witzel C. Fibrin glue repair leads to enhanced axonal elongation during early peripheral nerve regeneration in an in vivo mouse model. Neural Regen Res 2015; 10 (7) 1166-1171
  • 16 Feng G, Mellor RH, Bernstein M , et al. Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP. Neuron 2000; 28 (1) 41-51
  • 17 Koulaxouzidis G, Reutter W, Hildebrandt H, Stark GB, Witzel C. In vivo stimulation of early peripheral axon regeneration by N-propionylmannosamine in the presence of polysialyltransferase ST8SIA2. J Neural Transm (Vienna) 2015; 122 (9) 1211-1219
  • 18 Witzel C, Reutter W, Stark GB, Koulaxouzidis G. N-Propionylmannosamine stimulates axonal elongation in a murine model of sciatic nerve injury. Neural Regen Res 2015; 10 (6) 976-981
  • 19 Koulaxouzidis G, Reim G, Fluhr JW, Simunovic F, Stark GB, Witzel C. In Situ Deactivation of Interleukin-6 Enhances Early Peripheral Nerve Regeneration in a Murine Injury Model. J Reconstr Microsurg 2015; 31 (7) 508-515
  • 20 Fu SY, Gordon T. The cellular and molecular basis of peripheral nerve regeneration. Mol Neurobiol 1997; 14 (1–2) 67-116
  • 21 Brushart TM, Gerber J, Kessens P, Chen YG, Royall RM. Contributions of pathway and neuron to preferential motor reinnervation. J Neurosci 1998; 18 (21) 8674-8681
  • 22 Chen YG, Brushart TM. The effect of denervated muscle and Schwann cells on axon collateral sprouting. J Hand Surg Am 1998; 23 (6) 1025-1033
  • 23 Brushart TM. Motor axons preferentially reinnervate motor pathways. J Neurosci 1993; 13 (6) 2730-2738
  • 24 Mackinnon SE, Dellon AL, O'Brien JP. Changes in nerve fiber numbers distal to a nerve repair in the rat sciatic nerve model. Muscle Nerve 1991; 14 (11) 1116-1122
  • 25 Abdullah M, O'Daly A, Vyas A, Rohde C, Brushart TM. Adult motor axons preferentially reinnervate predegenerated muscle nerve. Exp Neurol 2013; 249: 1-7
  • 26 Fawcett JW, Schwab ME, Montani L, Brazda N, Müller HW. Defeating inhibition of regeneration by scar and myelin components. Handb Clin Neurol 2012; 109: 503-522
  • 27 Kadoya K, Tsukada S, Lu P , et al. Combined intrinsic and extrinsic neuronal mechanisms facilitate bridging axonal regeneration one year after spinal cord injury. Neuron 2009; 64 (2) 165-172
  • 28 Wyatt LA, Filbin MT, Keirstead HS. PTEN inhibition enhances neurite outgrowth in human embryonic stem cell-derived neuronal progenitor cells. J Comp Neurol 2014; 522 (12) 2741-2755
  • 29 Hannila SS, Filbin MT. The role of cyclic AMP signaling in promoting axonal regeneration after spinal cord injury. Exp Neurol 2008; 209 (2) 321-332
  • 30 Aglah C, Gordon T, Posse de Chaves EI. cAMP promotes neurite outgrowth and extension through protein kinase A but independently of Erk activation in cultured rat motoneurons. Neuropharmacology 2008; 55 (1) 8-17
  • 31 Gordon T. The role of neurotrophic factors in nerve regeneration. Neurosurg Focus 2009; 26 (2) E3
  • 32 Udina E, Furey M, Busch S, Silver J, Gordon T, Fouad K. Electrical stimulation of intact peripheral sensory axons in rats promotes outgrowth of their central projections. Exp Neurol 2008; 210 (1) 238-247
  • 33 Singh B, Xu QG, Franz CK , et al. Accelerated axon outgrowth, guidance, and target reinnervation across nerve transection gaps following a brief electrical stimulation paradigm. J Neurosurg 2012; 116 (3) 498-512
  • 34 Inalöz SS, Ak HE, Vayla V , et al. Comparison of microsuturing to the use of tissue adhesives in anastomosing sciatic nerve cuts in rats. Neurosurg Rev 1997; 20 (4) 250-258
  • 35 Martins RS, Siqueira MG, Silva CF, Godoy BO, Plese JP. Electrophysiologic assessment of regeneration in rat sciatic nerve repair using suture, fibrin glue or a combination of both techniques. Arq Neuropsiquiatr 2005; 63 (3A, 3a) 601-604
  • 36 Menovsky T, Beek JF. Laser, fibrin glue, or suture repair of peripheral nerves: a comparative functional, histological, and morphometric study in the rat sciatic nerve. J Neurosurg 2001; 95 (4) 694-699
  • 37 Ornelas L, Padilla L, Di Silvio M , et al. Fibrin glue: an alternative technique for nerve coaptation—Part I. Wave amplitude, conduction velocity, and plantar-length factors. J Reconstr Microsurg 2006; 22 (2) 119-122
  • 38 Ornelas L, Padilla L, Di Silvio M , et al. Fibrin glue: an alternative technique for nerve coaptation—Part II. Nerve regeneration and histomorphometric assessment. J Reconstr Microsurg 2006; 22 (2) 123-128