Abstract
The C5-C6 nerve roots are usually spared from avulsion after brachial plexus injury (BPI) and can thus be used as donors for nerve repair. A BPI rat model with C5-C6 nerve root stumps has been established in our previous work. The aim of this study was to test whether riluzole loaded into a thermosensitive hydrogel could applied locally in the nerve root stumps of this BPI rat model, thus increasing the reparative effect of the nerve root stumps. Nile red (a hydrophobic dye) was used as a substitute for riluzole since riluzole itself does not emit light. Nile red, loaded into a thermosensitive hydrogel, was added to the nerve root stumps of the BPI rat model. Additionally, eighteen rats, with operation on right brachial plexus, were evenly divided into three groups: control (Con), thermosensitive hydrogel (Gel) and thermosensitive hydrogel loaded with riluzole (Gel + Ri) groups. Direct nerve repair was performed after local riluzole release for two weeks. Functional and electrophysiological evaluations and histological assessments were used to evaluate the reparative effect 8 weeks after nerve repair. Nile red was slowly released from the thermosensitive hydrogel and retrograde transport through the nerve root stumps to the motoneurons, according to immunofluorescence. Discernible functional recovery began earlier in the Gel + Ri group. The compound muscle action potential, ChAT-expressing motoneurons, positivity for neurofilaments and S100, diameter of regenerating axons, myelin sheath thickness and density of myelinated fibers were markedly increased in the Gel + Ri group compared with the Con and Gel groups. Our results indicate that the local administration of riluzole could undergo retrograde transportation through C5-C6 nerve root stumps, thereby promoting neuroprotection and increasing nerve regeneration.
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References
Estrella EP (2011) Functional outcome of nerve transfers for upper-type brachial plexus injuries. J Plast Reconstr Aesthet Surg 64:1007–1013
Carlstedt T (2009) Nerve root replantation. Neurosurg Clin N Am 20:39–50
Sunderland S (1974) Mechanisms of cervical nerve root avulsion in injuries of the neck and shoulder. J Neurosurg 41:705–714
Addosooki A, Doi K, Hattori Y, Moriya A, Estrella E (2008) Evaluation of C5 nerve root repairability in traumatic brachial plexus injuries: proposal of an evaluation scoring system. J Reconstr Microsurg 24:3–10
Bertelli JA, Ghizoni MF (2008) Results of grafting the anterior and posterior divisions of the upper trunk in complete palsies of the brachial plexus. J Hand Surg Am 33:1529–1540
Kline DG (2009) Timing for brachial plexus injury: a personal experience. Neurosurg Clin N Am 20:24–26
Koliatsos VE, Price WL, Pardo CA, Price DL (1994) Ventral root avulsion: an experimental model of death of adult motor neurons. J Comp Neurol 342:35–44
Ygge J (1989) Neuronal loss in lumbar dorsal root ganglia after proximal compared to distal sciatic nerve resection: a quantitative study in the rat. Brain Res 478:193–195
Holtzer CA, Feirabend HK, Marani E, Thomeer RT (2000) Ultrastructural and quantitative motoneuronal changes after ventral root avulsion favor early surgical repair. Arch Physiol Biochem 108:293–309
Bensimon G, Lacomblez L, Meininger V (1994) A controlled trial of riluzole in amyotrophic lateral sclerosis. ALS/Riluzole Study Group. N Engl J Med 330:585–591
Lacomblez L, Bensimon G, Leigh PN, Guillet P, Meininger V (1996) Dose-ranging study of riluzole in amyotrophic lateral sclerosis. Amyotrophic Lateral Sclerosis/Riluzole Study Group II. Lancet 347:1425–1431
Doble A (1996) The pharmacology and mechanism of action of riluzole. Neurology 47:S233–S241
Srinivas S, Wali AR, Pham MH (2019) Efficacy of riluzole in the treatment of spinal cord injury: a systematic review of the literature. Neurosurg Focus 46:E6
Shimizu EN, Seifert JL, Johnson KJ, Romero-Ortega MI (2018) Prophylactic Riluzole Attenuates Oxidative Stress Damage in Spinal Cord Distraction. J Neurotrauma 35:1319–1328
Nagoshi N, Nakashima H, Fehlings MG (2015) Riluzole as a neuroprotective drug for spinal cord injury: from bench to bedside. Molecules 20:7775–7789
Nógrádi A, Szabó A, Pintér S, Vrbová G (2007) Delayed riluzole treatment is able to rescue injured rat spinal motoneurons. Neuroscience 144:431–438
Pintér S, Gloviczki B, Szabó A, Márton G, Nógrádi A (2010) Increased Survival and Reinnervation of Cervical Motoneurons by Riluzole after Avulsion of the C7 Ventral Root. J Neurotrauma 27:2273–2282
Fang J, Yang J, Yang Y, Li L, Qin B, He W, Yan L, Chen G, Tu Z, Liu X, Gu L (2018) A novel rat model of brachial plexus injury with nerve root stumps. J Neurosci Methods 295:1–9
Kharkar PM, Kiick KL, Kloxin AM (2013) Designing degradable hydrogels for orthogonal control of cell microenvironments. Chem Soc Rev 42:7335–7372
Pakulska MM, Ballios BG, Shoichet MS (2012) Injectable hydrogels for central nervous system therapy. Biomed Mater 7:24101
Klouda L (2015) Thermoresponsive hydrogels in biomedical applications: A seven-year update. Eur J Pharm Biopharm 97:338–349
Ruel-Gariepy E, Leroux JC (2004) In situ-forming hydrogels–review of temperature-sensitive systems. Eur J Pharm Biopharm 58:409–426
Tang S, Zhao J, Xu S, Li J, Teng Y, Quan D, Guo X (2012) Bone induction through controlled release of novel BMP-2-related peptide from PTMC(1)(1)-F127-PTMC(1)(1) hydrogels. Biomed Mater 7:15008
Wang T, Yan M, Sun X, Quan D (2015) The mechanical and biological properties of polycarbonate-modified F127 hydrogels after incorporating active pendent double-bonds. Polymer 57:21–28
Nicholson KJ, Zhang S, Gilliland TM, Winkelstein BA (2014) Riluzole effects on behavioral sensitivity and the development of axonal damage and spinal modifications that occur after painful nerve root compression. J Neurosurg Spine 20:751–762
Jiang K, Zhuang Y, Yan M, Chen H, Ge AQ, Sun L, Miao B (2016) Effects of riluzole on P2 × 7R expression in the spinal cord in rat model of neuropathic pain. Neurosci Lett 618:127–133
Gloviczki B, Török DG, Márton G, Gál L, Bodzay T, Pintér S, Nógrádi A (2017) Delayed Spinal Cord–Brachial Plexus Reconnection after C7 Ventral Root Avulsion: The Effect of Reinnervating Motoneurons Rescued by Riluzole Treatment. J Neurotrauma 34:2364–2374
Bertelli JA, Mira JC (1993) Behavioral evaluating methods in the objective clinical assessment of motor function after experimental brachial plexus reconstruction in the rat. J Neurosci Methods 46:203–208
Lucchi C, Curia G, Vinet J, Gualtieri F, Bresciani E, Locatelli V, Torsello A, Biagini G (2013) Protective but not anticonvulsant effects of ghrelin and JMV-1843 in the pilocarpine model of Status epilepticus. PLoS One 8:e72716
Bozkurt A, Lassner F, O’Dey D, Deumens R, Bocker A, Schwendt T, Janzen C, Suschek CV, Tolba R, Kobayashi E, Sellhaus B, Tholl S, Eummelen L, Schugner F, Damink LO, Weis J, Brook GA, Pallua N (2012) The role of microstructured and interconnected pore channels in a collagen-based nerve guide on axonal regeneration in peripheral nerves. Biomaterials 33:1363–1375
Gulino R (2016) Neuroplasticity and Repair in Rodent Neurotoxic Models of Spinal Motoneuron Disease. Neural Plast 2016:2769735
Wiley RG, Kline IR (2000) Neuronal lesioning with axonally transported toxins. J Neurosci Methods 103:73–82
Liu YS, Wang ZQ, Guo LH, Zhang TG, Sun SZ (2003) [The effect of adriamycin to the trigeminal ganglion following injection into the rabbits infraorbital nerve]. Shanghai Kou Qiang Yi Xue 12:447–452
Kato S, Otsuki T, Yamamoto T, Iwasaki Y, Yoshimoto T (1990) Retrograde adriamycin sensory ganglionectomy: novel approach for the treatment of intractable pain. Stereotact Funct Neurosurg 54–55:86–89
Huang J, Lin C, Fang J, Li X, Wang J, Deng S, Zhang S, Su W, Feng X, Chen B, Cheng D, Shuai X (2018) pH-Sensitive Nanocarrier-Mediated Codelivery of Simvastatin and Noggin siRNA for Synergistic Enhancement of Osteogenesis. ACS Appl Mater Interfaces 10:28471–28482
Faroni A, Mobasseri SA, Kingham PJ, Reid AJ (2015) Peripheral nerve regeneration: experimental strategies and future perspectives. Adv Drug Deliv Rev 82–83:160–167
Gu Y, Spasic Z, Wu W (1997) The effects of remaining axons on motoneuron survival and NOS expression following axotomy in the adult rat. Dev Neurosci 19:255–259
Burnett MG, Zager EL (2004) Pathophysiology of peripheral nerve injury: a brief review. Neurosurg Focus 16:E1
Spejo AB, Carvalho JL, Goes AM, Oliveira AL (2013) Neuroprotective effects of mesenchymal stem cells on spinal motoneurons following ventral root axotomy: synapse stability and axonal regeneration. Neuroscience 250:715–732
Zhao S, Pang Y, Beuerman RW, Thompson HW, Kline DG (1998) Expression of c-Fos protein in the spinal cord after brachial plexus injury: comparison of root avulsion and distal nerve transection. Neurosurgery 42:1357–1362, 1362–1363
Ma J, Novikov LN, Wiberg M, Kellerth JO (2001) Delayed loss of spinal motoneurons after peripheral nerve injury in adult rats: a quantitative morphological study. Exp Brain Res 139:216–223
Spejo AB, Oliveira AL (2015) Synaptic rearrangement following axonal injury: Old and new players. Neuropharmacology 96:113–123
Nógrádi A, Vrbová G (2001) The effect of riluzole treatment in rats on the survival of injured adult and grafted embryonic motoneurons. Eur J Neurosci 13:113–118
Cabaj AM, Slawinska U (2012) Riluzole treatment reduces motoneuron death induced by axotomy in newborn rats. J Neurotrauma 29:1506–1517
Ghayour MB, Abdolmaleki A, Behnam-Rassouli M (2017) The effect of Riluzole on functional recovery of locomotion in the rat sciatic nerve crush model. Eur J Trauma Emerg Surg 43:691–699
Yang JT, Fang JT, Li L, Chen G, Qin BG, Gu LQ (2019) Contralateral C7 transfer combined with acellular nerve allografts seeded with differentiated adipose stem cells for repairing upper brachial plexus injury in rats. Neural Regen Res 14:1932–1940
Zhu Z, Zhou X, He B, Dai T, Zheng C, Yang C, Zhu S, Zhu J, Zhu Q, Liu X (2015) Ginkgo biloba extract (EGb 761) promotes peripheral nerve regeneration and neovascularization after acellular nerve allografts in a rat model. Cell Mol Neurobiol 35:273–282
Acknowledgements
This work was supported by the National Natural Science Foundation of China (Grant Numbers 81572130, 81601057, and 81871787) and the Natural Science Foundation of Guangdong Province (Grant Numbers 2019A1515012057, 2018A030310254,2015A030310350 and 2015A030313194).
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JY and LG carried out the concept, and JF, LL and HZ participated in the design of this study and performed the experiments and statistical analysis. These three authors contributed to this work equally and should be considered co-first authors. BQ, ES and MZ also performed experiments and collected important background information. DQ and HZ designed and provided the thermosensitive hydrogel. DQ, XL, JY and LG provided technical support. JF, LL and HZ drafted the paper. JY and LG revised the paper. All authors read and approved the final manuscript.
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All animal studies have been approved by The Institute Research Medical Ethics Committee of Sun Yat-Sen University (Application ID: [2016] 150).
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Fang, J., Li, L., Zhai, H. et al. Local Riluzole Release from a Thermosensitive Hydrogel Rescues Injured Motoneurons through Nerve Root Stumps in a Brachial Plexus Injury Rat Model. Neurochem Res 45, 2800–2813 (2020). https://doi.org/10.1007/s11064-020-03120-0
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DOI: https://doi.org/10.1007/s11064-020-03120-0