Long-term anodal block stimulation at sacral anterior roots promoted recovery of neurogenic bladder function in a rabbit model of complete spinal cord injury

A complete spinal cord injury model was established in experimental rabbits using the spinal cord clip compression method.Urodynamic examination was performed 2 weeks later to determine neurogenic bladder status.The rabbits were treated with anodal block stimulation at sacral anterior roots for 4 weeks.Electrical stimulation of sacral anterior roots improved urodynamic parameters of neurogenic bladder in rabbit models of complete spinal cord injury,effectively promoted urinary function,and relieved urinary retention.Immunohistochemistry results showed that a balance was achieved among expression of muscarinic receptor subunits M2,M3,ATP-gated ion channel P2X3 receptors,and β2-adrenergic receptor,and nerve growth factor expression decreased.These results suggested that long-term sacral anterior root stimulation of anodal block could be used to treat neurogenic bladder in a rabbit model of complete spinal cord injury.     

Lumbar and Sacral Nerve Root Stimulation (NRS) in the Treatment of Chronic Pain: A Novel Anatomic Approach and Neuro Stimulation Technique

Objective. The conventional technique used to stimulate the lumbar dermatomes is by stimulation of the dorsal columns of the spinal cord. Until recently, stimulation of nerve roots had not been successfully accomplished. We had performed selective nerve root cannulations for the placement of temporary catheters at cervical, thoracic, lumbar, and sacral levels in chronic pain patients using a caudad rather than craniad approach. We hypothesized that by stimulating the nerve roots we could improve paresthesia coverage in areas which cannot be covered effectively by spinal cord stimulation (SCS). To test this hypothesis, we have performed trials of nerve root stimulation (NRS) in patients who had failed SCS, or who were not candidates for SCS because their pain was otherwise inaccessible to stimulation. Methods. Five patients who had been unresponsive to conservative treatment, surgery, or SCS underwent 7-day trials with NRS. The diagnoses included: ilioinguinal neuralgia, discogenic low back pain, failed back syndrome, vulvodynia, and interstitial cystitis. We collected paresthesia maps, pain maps, pain visual analog scale (VAS) scores, and patient satisfaction ratings. Results. Paresthesia coverage was above 75% in all patients. VAS scores declined from a mean of 9 ± 1.0 to 2.4 ± 2.1 (p < 0.05, n= 5), all 5 patients requested permanent implantation, and 4 have been implanted so far. Conclusions. Lumbar and sacral NRS trials resulted in adequate paresthesia coverage and effective pain relief in all 5 patients. Further clinical trials to evaluate long-term success rates and safety are indicated. Detailed mapping studies are needed to evaluate the relationship between electrode placement and paresthesia patterns as well as the optimal stimulation parameters.     

The Inhibitory Effects of Pudendal Nerve Stimulation on Bladder Overactivity in Spinal Cord Injury Dogs: Is Early Stimulation Necessary?

Objective: To determine the inhibitory effects of pudendal nerve stimulation (5 Hz) on bladder overactivity at early and late stages of spinal cord injury in dogs. Materials and Methods: The study was performed in eight dogs with chronic spinal cord transection at the T9‐T10 level. Group 1 (four dogs) underwent electrical stimulation of pudendal nerve one month after spinal cord transection. Group 2 (four dogs) underwent stimulation six months after spinal cord transection. The bladders were removed for histological examination of fibrosis after the stimulation. Results: The bladder capacity and the compliance were significantly increased (p < 0.05) by pudendal nerve stimulation in group 1, but not in group 2. The nonvoiding contractions were inhibited in both groups by electrical stimulation. Collagen fiber was increased, while elastic fiber was significantly decreased (p < 0.05) in group 2 when compared with group 1. Conclusion: Pudendal nerve stimulation can increase the bladder capacity and compliance only during the early period before the bladder wall becomes fibrosit and can inhibit the nonvoiding contraction during two stages.     

Olfactory ensheathing cell grafts have minimal influence on regeneration at the dorsal root entry zone following rhizotomy

The effectiveness of grafts of olfactory ensheathing cells (OECs) as a means of promoting functional reconnection of regenerating primary afferent fibers was investigated following dorsal root injury. Adult rats were subjected to dorsal root section and reanastomosis and at the same operation a suspension of purified OECs was injected at the dorsal root entry zone and/or into the sectioned dorsal root. Regeneration of dorsal root fibers was then assessed after a survival period ranging from 1 to 6 months. In 11 animals, electrophysiology was used to look for evidence of functional reconnection of regenerating dorsal root fibers. However, electrical stimulation of lesioned dorsal roots failed to evoke detectable cord dorsum or field potentials within the spinal cord of any of the animals examined, indicating that reconnection of regenerating fibers with spinal cord neurones had not occurred. In a further 11 rats, immunocytochemical labeling and biotin dextran tracing of afferent fibers in the lesioned roots was used to determine whether regenerating fibers were able to grow into the spinal cord in the presence of an OEC graft. Although a few afferent fibers could be seen to extend for a limited distance into the spinal cord, similar minimal in-growth was seen in control animals that had not been injected with OECs. We therefore conclude that OEC grafts are of little or no advantage in promoting the in-growth of regenerating afferent fibers at the dorsal root entry zone following rhizotomy.     

Electrical stimulation of dog pudendal nerve regulates the excitatory pudendal-to-bladder relfex

Pudendal nerve plays an important role in urine storage and voiding. Our hypothesis is that a neuroprosthetic device placed in the puden-dal nerve trunk can modulate bladder function after suprasacral spinal cord injury. We had conifrmed the inhibitory pudendal-to-bladder relfex by stimulating either the branch or the trunk of the pudendal nerve. This study explored the excitatory pudendal-to-bladder relfex in beagle dogs, with intact or injured spinal cord, by electrical stimulation of the pudendal nerve trunk. The optimal stimulation frequency was approximately 15–25 Hz. This excitatory effect was dependent to some extent on the bladder volume. We conclude that stimulation of the pudendal nerve trunk is a promising method to modulate bladder function.     

Calcitonin gene-related peptide in anterior and posterior horns of spinal cord after brachial plexus injury

BACKGROUND: The changes of calcitonin gene-related peptide (CGRP) expression are closely associated with peripheral nerve injury, whereas it should be further investigated whether the damage of central nerve can lead to the changes of CGRP expression, and whether it is associated with the neural regeneration and repair. OBJECTIVE: To observe the changing law of CGRP expression in the anterior and posterior horns of spinal cord following brachial plexus injury. DESIGN: A randomized controlled trial. SETTINGS: Department of Anatomy, Yunyang Medical College; Department of Anatomy, Basic Medical College, Sun Yat-sen University. MATERIALS: Sixty-five adult male SD rats of clean degree, weighing 180–220 g, provided by the experimental animal center of the Basic Medical College, Sun Yat-sen University, were randomly divided into control group (n =5) and experimental group (n =60), and the latter was subdivided into three damage groups: avulsion of anterior root group (n =20), disjunction of posterior root group (n =20) and transection of spinal cord group (n =20). Diaminobenzidine (DAB) chromogen, rabbit anti-CGRP polyclonal antibody were the products of Sigma Company; Leica image analytical apparatus was produced by QUIN Company (Germany); Histotome by Sigma Company. METHODS: The experiments were carried out in the Department of Anatomy, Basic Medical College, Sun Yat-sen University from September 2004 to March 2005. Three kinds of models of brachial plexus injury were established: In the avulsion of anterior root group, right C7 anterior root was avulsed, and the distal nerve residual root was transected. In the disjunction of posterior root group, right C7 anterior root was avulsed and right C5–T1 posterior horns were cut to block the sensory afferent pathway. In the transection of spinal cord group, right C7 anterior root was avulsed and C5–6 segments of right spinal cord were semi-transected to block the cortical descending pathway. In the control group, C5–T1 vertebral plates were prayed open, and then the skin was sutured. The C7 segments of spinal cord were removed on the 1st, 3rd, 7th and 14th days postoperatively respectively, and the CGRP expressions in the anterior and posterior horns of spinal cord were determined and analyzed using immunohistochemical method and image analysis. MAIN OUTCOME MEASURES: ① Number of CGRP immuno-positive motor neurons in the anterior horn of spinal cord; ② Total area of CGRP immuno-positive fibers in the posterior horn of spinal cord. RESULTS: All the 65 rats were involved in the analysis of results. ① Number of CGRP immuno-positive motor neurons in the anterior horn of spinal cord: CGRP immuno-positive motor neurons could be observed in the anterior horns of C7 spinal cord in the control group and damage groups, the neurons had big cell body with stained cytoplasm, appeared as brown granules, and mainly distributed in the ventral lateral anterior horn of spinal cord. On the 1st day postoperatively, the number of CGRP positive neurons was obviously higher in the in the avulsion of anterior root group than in the control group (P < 0.01), whereas obviously lower in the disjunction of posterior root group than in the control group (P < 0.01), and there was no obvious difference between the transection of spinal cord group and the control group (P > 0.05). On the 7th day, the numbers of CGRP positive neurons in the damage groups were obviously higher than that in the control group (P < 0.01), also obviously different from those on the 1st day in the same group respectively (P < 0.01). On the 14th day, the number of CGRP positive neurons in the disjunction of posterior root group was decreased, but there was no obvious difference as compared with that in the control group, whereas those in the avulsion of anterior root group and transection of spinal cord group were still obviously higher than that in the control group (P < 0.01). The number of CGRP positive neurons was the most in the avulsion of anterior root group, followed by the transection of spinal cord group, and the least in the disjunction of posterior root group, and there were significant differences among them (P < 0.01). ② Total area of CGRP immuno-positive fibers in the posterior horn of spinal cord: Dense CGRP immuno-positive nerve fibers distributed in the layers Ⅰ and Ⅱ of the C7 posterior horn of spinal cord in the control group. On the 1st day postoperatively, the total areas of CGRP positive fibers in the avulsion of anterior root group and transection of spinal cord group were obviously larger than that in the control group (P < 0.01), whereas there was no obvious difference between the disjunction of posterior root group and control group. On the 7th day, the CGRP expression in the posterior horn of spinal cord decreased to the lowest level in the disjunction of posterior root group, whereas there were no obvious differences in the avulsion of anterior root group and transection of spinal cord group as compared with that in the control group (P > 0.05). On the 14th day, the area continued to decrease in the avulsion of anterior root group and transection of spinal cord group, and it was obviously lower in the transection of spinal cord group than in the control group (P < 0.01), and it was slightly increased in the disjunction of posterior root group as compared with that on 7th day, but still obviously lower than that in the control group (P < 0.01). CONCLUSION: The expression and role of CGRP are in discrepancy in the anterior and posterior horns of spinal cord after brachial plexus injury. The CGRP in anterior horn of spinal cord are derived from the cell body of motor neurons, and may be involved in the repairing mechanism of nerve injury regeneration; Whereas those in the posterior horn are mainly derived from posterior root ganglion, and may be associated with the conduction of noxious stimulations.     

The effect of peripheral nerve injury on dorsal root potentials and on transmission of afferent signals into the spinal cord

The sciatic nerve adults rats was either cut and ligated or was crushed on one side. The response of the spinal cord to stimulation of the proximal part of the injured nerve was examined at various times after the lesion and compared to the effects of stimulating the intact nerve on the other side. During the first 10 days after nerve section the following measures were not affected: (i) the size of the input volley (compound action potential, CAP, measured on a dorsal root that carried sciatic nerve afferents (L5); (ii) the volley running in the dorsal columns; (iii) the dorsal root potential (DRP) evoked on neighbouring dorsal roots which do not contain sciatic afferents (L2 and L3); (iv) the post-synaptic volleys ascending in the spinal cord. However, by the fourth day after nerve section, there was a decrease of the DRP evoked on the ipsilateral L5 dorsal root by stimulation of the cut nerve. By 10 days this DRP had decreased by 50%. There was also a decrease in the DRP on the L5 root evoked by stimulation of the contralateral intact nerve. Crush lesions of the sciatic nerve did not produce DRP charge. Beginning 10–20 days after nerve cut, there was a decrease in the amplitude of the afferent CAP and of all the measures of central response to the afferent volley. We discuss the possibility that the loss of the DRP may be associated with a disinhibition which results in novel receptive fields which we observe in cord cells deafferented by the peripheral nerve section. The decrease of DRP and the appearance of novel receptive fields do not occur if the peripheral nerve is crushed rather than cut.     

Functional repair after dorsal root rhizotomy using nerve conduits and neurotrophic molecules

Functional recovery after large excision of dorsal roots is absent because of both the limited regeneration capacity of the transected root, and the inability of regenerating sensory fibers to traverse the dorsal root entry zone. In this study, bioresorbable guidance conduits were used to repair 6-mm dorsal root lesion gaps in rats, while neurotrophin-encoding adenoviruses were used to elicit regeneration into the spinal cord. Polyester conduits with or without microfilament bundles were implanted between the transected ends of lumbar dorsal roots. Four weeks later, adenoviruses encoding NGF or GFP were injected into the spinal cord along the entry zone of the damaged dorsal roots. Eight weeks after injury, nerve regeneration was observed through both types of implants, but those containing microfilaments supported more robust regeneration of calcitonin gene-related peptide (CGRP)-positive nociceptive axons. NGF overexpression induced extensive regeneration of CGRP(+) fibers into the spinal cord from implants showing nerve repair. Animals that received conduits containing microfilaments combined with spinal NGF virus injections showed the greatest recovery in nociceptive function, approaching a normal level by 7-8 weeks. This recovery was reversed by recutting the dorsal root through the centre of the conduit, demonstrating that regeneration through the implant, and not sprouting of intact spinal fibers, restored sensory function. This study demonstrates that a combination of PNS guidance conduits and CNS neurotrophin therapy can promote regeneration and restoration of sensory function after severe dorsal root injury.     

Rebuilding motor function of the spinal cord based on functional electrical stimulation

Rebuilding the damaged motor function caused by spinal cord injury is one of the most serious challenges in clinical neuroscience. The function of the neural pathway under the damaged sites can be rebuilt using functional electrical stimulation technology. In this study, the locations of motor function sites in the lumbosacral spinal cord were determined with functional electrical stimulation technology. A three-dimensional map of the lumbosacral spinal cord comprising the relationship between the motor function sites and the correspond-ing muscle was drawn. Based on the individual experimental parameters and normalized coordinates of the motor function sites, the motor function sites that control a certain muscle were calculated. Phasing pulse sequences were delivered to the determined motor function sites in the spinal cord and hip extension, hip lfexion, ankle plantarlfexion, and ankle dorsilfexion movements were successfully achieved. The results show that the map of the spinal cord motor function sites was valid. This map can provide guidance for the selection of electrical stimulation sites during the rebuilding of motor function after spinal cord injury.     

Interfacing peripheral nerve with macro-sieve electrodes following spinal cord injury

Macro-sieve electrodes were implanted in the sciatic nerve of five adult male Lewis rats following spinal cord injury to assess the ability of the macro-sieve electrode to interface regenerated peripheral nerve fibers post-spinal cord injury. Each spinal cord injury was performed via right lateral hemisection of the cord at the T_(9–10) site. Five months post-implantation, the ability of the macro-sieve electrode to interface the regenerated nerve was assessed by stimulating through the macro-sieve electrode and recording both electromyography signals and evoked muscle force from distal musculature. Electromyography measurements were recorded from the tibialis anterior and gastrocnemius muscles, while evoked muscle force measurements were recorded from the tibialis anterior, extensor digitorum longus, and gastrocnemius muscles. The macro-sieve electrode and regenerated sciatic nerve were then explanted for histological evaluation. Successful sciatic nerve regeneration across the macro-sieve electrode interface following spinal cord injury was seen in all five animals. Recorded electromyography signals and muscle force recordings obtained through macro-sieve electrode stimulation confirm the ability of the macro-sieve electrode to successfully recruit distal musculature in this injury model. Taken together, these results demonstrate the macro-sieve electrode as a viable interface for peripheral nerve stimulation in the context of spinal cord injury.     

Glial cell reactions in the spinal cord after sensory nerve stimulation are associated with axonal injury

Astroglial and microglial reactions in the dorsal and ventral horns of the adult rat spinal cord were studied after graded electrical stimulation of the rat sciatic nerve and after topical application of mustard oil to the hindlimb foot. Antibodies to glial fibrillary acidic protein and complement receptor 3 (OX-42) were used as markers for astroglia and microglia, respectively. The results showed that electrical nerve stimulation resulted in increased immunoreactivity for GFAP and OX-42 in the spinal cord dorsal and ventral horns only after the use of stimulation strengths which were associated with nerve fiber degeneration in the stimulated nerve. Application of mustard oil to the foot caused no changes in GFAP or OX-42 immunoreactivity. These findings indicate that peripheral nerve stimulation in itself is insufficient to induce astroglial and microglial responses in the spinal cord. The signal(s) mediating these responses, regularly seen after nerve injury, are therefore most probably not related to the afferent barrage of action potentials evoked by the injury.     

Microtubule stabilization promoted axonal regeneration and functional recovery after spinal root avulsion

A spinal root avulsion injury disconnects spinal roots with the spinal cord. The rampant motoneuron death, inhibitory CNS/PNS transitional zone (TZ) for axonal regrowth and limited regeneration speed together lead to motor dysfunction. Microtubules rearrange to assemble a new growth cone and disorganized microtubules underline regeneration failure. It has been shown that microtubule‐stabilizing drug, Epothilone B, enhanced axonal regeneration and attenuated fibrotic scaring after spinal cord injury. Here, we are reporting that after spinal root avulsion+ re‐implantation in adult rats, EpoB treatment improved motor functional recovery and potentiated electrical responses of motor units. It facilitated axons to cross the TZ and promoted more and bigger axons in the peripheral nerve. Neuromuscular junctions were reformed with better preserved postsynaptic structure, and muscle atrophy was prevented by EpoB administration. Our study showed that EpoB was a promising therapy for promoting axonal regeneration after peripheral nerve injury.     

Epidural electrical stimulation effectively restores locomotion function in rats with complete spinal cord injury

Epidural electrical stimulation can restore limb motor function after spinal cord injury by reactivating the surviving neural circuits.In previous epidural electrical stimulation studies,single electrode sites and continuous tetanic stimulation have often been used.With this stimulation,the body is prone to declines in tolerance and locomotion coordination.In the present study,rat models of complete spinal cord injury were established by vertically cutting the spinal cord at the T8 level to eliminate disturbance from residual nerve fibers,and were then subjected to epidural electrical stimulation.The flexible extradural electrode had good anatomical topology and matched the shape of the spinal canal of the implanted segment.Simultaneously,the electrode stimulation site was able to be accurately applied to the L2–3 and S1 segments of the spinal cord.To evaluate the biocompatibility of the implanted epidural electrical stimulation electrodes,GFAP/Iba-1 doublelabeled immunofluorescence staining was performed on the spinal cord below the electrodes at 7 days after the electrode implantation.Immunofluorescence results revealed no significant differences in the numbers or morphologies of microglia and astrocytes in the spinal cord after electrode implantation,and there was no activated Iba-1~+ cell aggregation,indicating that the implant did not cause an inflammatory response in the spinal cord.Rat gait analysis showed that,at 3 days after surgery,gait became coordinated in rats with spinal cord injury under burst stimulation.The regained locomotion could clearly distinguish the support phase and the swing phase and dynamically adjust with the frequency of stimulus distribution.To evaluate the matching degree between the flexible epidural electrode(including three stimulation contacts),vertebral morphology,and the level of the epidural site of the stimulation electrode,micro-CT was used to scan the thoracolumbar vertebrae of rats before and after electrode implantation.Based on the experimental results of gait recovery using three-site stimulation electrodes at L2–3 and S1 combined with burst stimulation in a rat model of spinal cord injury,epidural electrical stimulation is a promising protocol that needs to be further explored.This study was approved by the Animal Ethics Committee of Chinese PLA General Hospital(approval No.2019-X15-39) on April 19,2019.     

Pulsed electrical stimulation protects neurons in the dorsal root and anterior horn of the spinal cord after peripheral nerve injury

Most studies on peripheral nerve injury have focused on repair at the site of injury, but very few have examined the effects of repair strategies on the more proximal neuronal cell bodies. In this study, an approximately 10-mm-long nerve segment from the ischial tuberosity in the rat was transected and its proximal and distal ends were inverted and sutured. The spinal cord was subjected to pulsed electrical stimulation at T10 and L3, at a current of 6.5 m A and a stimulation frequency of 15 Hz, 15 minutes per session, twice a day for 56 days. After pulsed electrical stimulation, the number of neurons in the dorsal root ganglion and anterior horn was increased in rats with sciatic nerve injury. The number of myelinated nerve fibers was increased in the sciatic nerve. The ultrastructure of neurons in the dorsal root ganglion and spinal cord was noticeably improved. Conduction velocity of the sciatic nerve was also increased. These results show that pulsed electrical stimulation protects sensory neurons in the dorsal root ganglia as well as motor neurons in the anterior horn of the spinal cord after peripheral nerve injury, and that it promotes the regeneration of peripheral nerve fibers.     

Sacral Nerve and Spinal Cord Stimulation for Intractable Neuropathic Pain Caused by Spinal Cord Infarction

Central cord pain is very difficult to relieve, even with the many kinds of medical and surgical treatments available. Following spinal cord infarctions, central cord pain can develop. The problems that may arise could include limb pain, pelvic pain, difficulties voiding, and difficulties defecating. We are reporting a case of central cord pain caused by a spinal cord infarction of the conus medullaris. Limb pain was reduced by spinal cord stimulation. Voiding and defecation difficulties and pelvic pain were reduced by sacral nerve stimulation. Thus, in a case involving both intractable limb and pelvic pain, a combination therapy of these two stimulations might be an effective treatment modality.     

Impulse magnetic stimulation facilitates synaptic regeneration in rats following sciatic nerve injury

The current studies describing magnetic stimulation for treatment of nervous system diseases mainly focus on transcranial magnetic stimulation and rarely focus on spinal cord magnetic stimula-tion.Spinal cord magnetic stimulation has been confirmed to promote neural plasticity after injuries of spinal cord,brain and peripheral nerve.To evaluate the effects of impulse magnetic stimulation of the spinal cord on peripheral nerve regneration,we compressed a 3 mm segment located in the middle third of the hip using a sterilized artery forceps to induce ischemia.Then,all animals un-derwent impulse magnetic stimulation of the lumbar portion of spinal crod and spinal nerve roots daily for 1 month.Electron microscopy results showed that in and below the injuryed segment,the inflammation and demyelination of neural tissue were alleviated,apoptotic cells were reduced,and injured Schwann cells and myelin fibers were repaired.These findings suggest that high-frequency impulse magnetic stimulation of spinal cord and corresponding spinal nerve roots promotes synaptic regeneration following sciatic nerve injury.     

Differential decerebrate control of depolarization in the central terminals of cutaneous afferents in the sacral cord

Presynaptic depolarization of cutaneous afferents has been investigated in the sacral cord of decerebrate cats before and after spinal cord transection. In the decerebrate state the central terminals of caudal femoral cutaneous nerve are depolarized by ipsilateral volleys entering the cord via sacral and lumbar dorsal roots. A significant increase of depolarization occurring after severing the cord indicates that there is tonic decerebrate inhibition of presynaptic depolarization in terminals of caudal femoral cutaneous nerve. In contrast to this finding, presynaptic depolarization evoked in the central terminals of the pudendal nerve by ipsilateral volleys entering the cord through sacral and lumbar dorsal roots is not subjected to decerebrate inhibitory control. It is suggested that differential inhibitory control of depolarization in the central terminals of cutaneous nerves in the sacral cord is related to the intraspinal course of their fibres, to differences in the receptor types involved, and to the location of their innervation fields. In more than half of the decerebrate preparations stimulation of the central terminals of cutaneous afferents through microelectrodes evokes antidromic spikes appearing simultaneously in ipsi- and contralateral nerves. The time course of bilateral excitability changes is similar on both sides of the cord. It is assumed that presynaptic effects are transmitted to the contralateral side by collaterals of ipsilateral cutaneous afferents.     

Synergistic actions of olomoucine and bone morphogenetic protein-4 in axonal repair after acute spinal cord contusion

To determine whether olomoucine acts synergistically with bone morphogenetic protein-4 in the treatment of spinal cord injury, we established a rat model of acute spinal cord contusion by impacting the spinal cord at the T8 vertebra. We injected a suspension of astrocytes derived from glial-restricted precursor cells exposed to bone morphogenetic protein-4 （GDAsBMP） into the spinal cord around the site of the injury, and/or olomoucine intraperitoneally. Olomoucine effectively inhibited astrocyte proliferation and the formation of scar tissue at the injury site, but did not prevent proliferation of GDAsBMP or inhibit their effects in reducing the spinal cord lesion cavity. Furthermore, while GDAsBMP and olomoucine independently resulted in small improve- ments in locomotor function in injured rats, combined administration of both treatments had a significantly greater effect on the restoration of motor function. These data indicate that the combined use of olomoucine and GDAsBMP creates a better environment for nerve regeneration than the use of either treatment alone, and contributes to spinal cord repair after injury.     

Oxidative phosphorylated neurofilament protein M protects spinal cord against ischemia/reperfusion injury

Previous studies have shown that neurofilament protein M expression is upregulated in the early stage of spinal cord ischemia/reperfusion injury, indicating that this protein may play a role in the injury process. In the present study, we compared protein expression in spinal cord tissue of rabbits after 25 minutes of ischemia followed by 0, 12, 24, or 48 hours of reperfusion with that of sham operated rabbits, using proteomic two-dimensional gel electrophoresis and mass spec- trometry. In addition, the nerve repair-related neurofilament protein M with the unregulated expression was detected with immunohistochemistry and western blot analysis. Two-dimen- sional gel electrophoresis and mass spectrometry showed that, compared with the sham group, upregulation of protein expression was most significant in the spinal cords of rabbits that had undergone ischemia and 24 hours of reperfusion. Immunohistochemical analysis revealed that neurofilament protein M was located in the membrane and cytoplasm of neuronal soma and axons at each time point after injury. Western blot analysis showed that neurofilament protein M expression increased with reperfusion time until it peaked at 24 hours and returned to baseline level after 48 hours. Furthermore, neurofilament protein M is phosphorylated under oxidative stress, and expression changes were parallel for the phosphorylated and non-phosphorylated forms. Neurofilament protein M plays an important role in spinal cord ischemia/reperfusion injury, and its functions are achieved through oxidative phosphorylation.     

Intranasal nerve growth factor bypasses the blood-brain barrier and affects spinal cord neurons in spinal cord injur y

The purpose of this work was to investigate whether, by intranasal administration, the nerve growth factor bypasses the blood-brain barrier and turns over the spinal cord neurons and if such therapeutic approach could be of value in the treatment of spinal cord injury. Adult Sprague-Dawley rats with intact and injured spinal cord received daily intranasal nerve growth factor administration in both nostrils for 1 day or for 3 consecutive weeks. We found an in-creased content of nerve growth factor and enhanced expression of nerve growth factor receptor in the spinal cord 24 hours after a single intranasal administration of nerve growth factor in healthy rats, while daily treatment for 3 weeks in a model of spinal cord injury improved the deifcits in locomotor behaviour and increased spinal content of both nerve growth factor and nerve growth factor receptors. These outcomes suggest that the intranasal nerve growth factor bypasses blood-brain barrier and affects spinal cord neurons in spinal cord injury. They also suggest exploiting the possible therapeutic role of intranasally delivered nerve growth factor for the neuroprotection of damaged spinal nerve cells.