Masahito Hara

Intravenously transplanted human neural stem cells migrate to the injured spinal cord in adult mice in an SDF-1- and HGF-dependent manner.

Neural stem cell (NSC) transplantation has exhibited considerable therapeutic potential in spinal cord injury. However, most experiments in animals have been performed by injecting these cells directly into the injured spinal cord. A cardinal feature of NSCs is their exceptional migratory ability through the nervous system. Based on the migratory ability of NSCs, we investigated whether minimally invasive intravenous delivery of NSCs could facilitate their migration to the injured spinal cord and identified the chemo-attractants secreted by the lesions. Nude mice were injected intravenously with labelled human NSCs at 3, 7 and 10 days after the compression of the spinal cord at the T8 level. The migration of NSCs to the lesioned spinal cord was highest at 7 days after injury; this correlated with the peak of hepatocyte growth factor and stromal cell-derived factor-1 mRNA expressions in the lesion but not with the disruption of the blood-brain barrier. Finally, the grafted NSCs differentiated into neuronal and glial subpopulations at 21 days after transplantation. Our study suggests that intravenously administered NSCs can be employed as a renewable source for replacing lost cells for the treatment of spinal cord injuries.

Activation of JAK/STAT signalling in neurons following spinal cord injury in mice

The Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signalling pathway is one of the most important in transducing signals from the cell surface to the nucleus in response to cytokines. In the present study, we investigated chronological alteration and cellular location of JAK1, STAT3, phosphorylated (p)‐Tyr1022/1023‐JAK1, p‐Tyr705‐STAT3, and interleukin‐6 (IL‐6) following spinal cord injury (SCI) in mice. Western blot analysis showed JAK1 to be significantly phosphorylated at Tyr1022/1023 from 6 h after SCI, peaking at 12 h and gradually decreasing thereafter, accompanied by phosphorylation of STAT3 at Tyr705 with a similar time course. ELISA analysis showed the concentration of IL‐6 in injured spinal cord to also significantly increase from 3 h after SCI, peaking at 12 h, then gradually decreasing. Immunohistochemistry revealed p‐Tyr1022/1023‐JAK1, p‐Tyr705‐STAT3, and IL‐6 to be mainly expressed in neurons of the anterior horns at 12 h after SCI. Pretreatment with a JAK inhibitor, AG‐490, suppressed phosphorylation of JAK1 and STAT3 at 12 h after SCI, reducing recovery of motor functions. These findings suggest that SCI at the acute stage produces IL‐6 mainly in neurons of the injured spinal cord, which activates the JAK/STAT pathway, and that this pathway may be involved with neuronal response to SCI.

Adrenomedullin reduces ischemic brain injury after transient middle cerebral artery occlusion in rats.

Summary Background. The effect of adrenomedullin, a vasodilatory peptide on transient middle cerebral artery (MCA) occlusion was investigated in rats. Methods. Transient MCA occlusion for 2 hours was made by using the intra-arterial suture method, followed by reperfusion. Findings. An intravenous infusion of adrenomedullin (1 μ g/kg/min) from one hour before ischemia to one hour after ischemia significantly reduced the infarct size and improved neurological deficits (p<0.05), without affecting systemic blood pressure or other physiological parameters. The infarct size was reduced with adrenomedullin by 25.4±12.7%, 31.3±5.8%, 31.6±6.1% respectively at the coronal level 6, 8 and 10 mm posterior from the frontal pole. Adrenomedullin also significantly inhibited the increase in myeloperoxidase (MPO) activity in the MCA area of the ischemic hemisphere after 22-hour reperfusion (control: 0.205±0.054 unit/g wet tissue, adrenomedullin group: 0.047±0.009 unit/g wet tissue, p<0.0001). Interpretation. These data suggest that adrenomedullin reduces acute ischemic brain injury and one of is neuroprotective mechanisms may be derived from inhibition of the infiltration of neutrophils into the ischemic tissue.

En Bloc Laminoplasty Performed with Threadwire Saw: Technical Note

OBJECTIVE To introduce a method for a simple, nonexpansive laminoplasty that can be performed with a threadwire saw (T-saw) after en bloc laminotomy has been performed. The method can be applied along the entire spinal region, including the thoracic and lumbar spine. METHODS An en bloc laminotomy of trapezoid shape at the cross section is performed bilaterally at the junctional area of the lamina and facet joint with a thin, flexible T-saw, while preserving the supraspinous, interspinous, and interlaminar ligaments. After the intradural procedure has been performed, the laminar flap is replaced in its original site and fixed with 1-0 nylon sutures, resulting in the complete reconstruction of the posterior supporting elements of the spinal column. RESULTS En bloc laminoplasty was performed on 16 patients via a T-saw; most of the patients had intradural spinal tumors. The patients did not need their spinal canals to be enlarged after the intradural procedure had been performed. The follow-up period ranged from 2 to 40 months (mean ± standard error, 22.6 ± 3.4 mo). The laminoplasty was performed from the upper cervical to the sacral regions, although the most frequently operated level was the lower thoracic level. Two-level laminoplasty was performed in 12 patients, and three-level laminoplasty was performed in four. The laminoplasty was done safely and without any complications, except in one patient, who experienced thoracic root injury from a T-saw that was accidentally inserted anterior to the roots. No spinal column deformity or sinking of the replaced laminar flap was noted during the follow-up period; patients were assessed at follow-up via radiographs or computed tomographic scans. Computed tomographic scans obtained later indicated that bony fusion occurred at the cutting edges 1.0 to 4.0 months after surgery (mean, 1.90 ± 0.34 mo). CONCLUSION Simple en bloc laminoplasty performed with a T-saw is a useful, safe procedure that can be used to reconstruct the posterior spinal elements throughout the whole spinal region after the intradural procedure has been performed.

A simple technique for expansive suboccipital cranioplasty following foramen magnum decompression for the treatment of syringomyelia associated with Chiari I malformation

The benefits of osteoplastic suboccipital craniotomies over the traditional suboccipital craniectomies have been recognized. We describe a simple method of expansive suboccipital cranioplastic craniotomy using a free bone flap and report satisfactory clinical results in 16 patients with syringomyelia associated with Chiari I malformation. A free suboccipital bone flap is created from the rostral part of the occiput by placing two to four burr holes and connecting them with a craniotome. The posterior bony margin of the foramen magnum and the posterior arch of C1 are removed thereafter. Then dural plasty using a patch graft of dural substitutes is performed. The expansive suboccipital cranioplasty is performed by positioning the free bone flap caudal to the original location and fixing it with titanium miniplates to construct a bony frame to cover the foramen magnum. The rostral part of the cranial defect is filled with bone chips created during the craniotomy. Sixteen patients underwent this procedure. There was no operative mortality and no major complication, such as persistent pseudomeningocele. Preoperative symptoms improved significantly in all patients except for one who had persistent dysesthetic pain. Our simple method of expansive suboccipital cranioplasty for the treatment of syringomyelia associated with Chiari I malformation proved useful and achieved satisfactory long-term results.

Taurine Reduces Inflammatory Responses after Spinal Cord Injury

Taurine has multiple functions in the central nervous system (CNS), serving as an osmoregulator, antioxidant, inhibitory neuromodulator, and regulator of intracellular Ca(2+) flux. Since the role of taurine in traumatic spinal cord injury (SCI) is not fully understood, the present study was conducted with C57 black/6 mice (18-20 g) who underwent severe SCI at the Th-8 level using a weight compression device. Taurine was injected intraperitoneally at doses of 25, 80, 250, and 800 mg/kg within 30 min after SCI. Controls were injected with saline. The contusional cord segments were removed 6 h after SCI, and concentrations of interleukin-6 (IL-6) and myeloperoxidase (MPO) were measured using ELISA kits. Phosphorylation of STAT3, which is activated by IL-6, and expression of inducible cyclooxygenase-2 (COX-2) were also compared between the taurine treatment group (250 mg/kg) and the control group by Western blot analysis. Morphological changes were evaluated with H&E-stained sections. Taurine significantly decreased IL-6 and MPO levels in a dose-dependent manner, significantly reducing the phosphorylation of STAT3 and expression of COX-2 after SCI compared to controls. A reduced accumulation of neutrophils, especially in the subarachnoid spaces, and secondary degenerative changes in gray matter were also noted, and motor disturbances were significantly attenuated with taurine treatment (250 mg/kg). These findings indicate that taurine has anti-inflammatory effects against SCI, and may play a neuroprotective role against secondary damage, and thus it may have therapeutic potential.

Anteriorty Cervical Decompression and Fusion for Cervical Spondylosis Using Vertebral Grafts Obtained from the Fusion site. Technical Advantages and Follow-up Results

Summary A retrospective analysis was done in 60 consecutive patients who underwent anterior cervical fusion using vertebral grafts obtained from the fusion site at the Nagoya University and its affiliated hospitals by a single surgeon (MT). Follow-up results and technical advantages are reported. The average follow-up period was 33 months (range 6 to 55 months). Sufficient decompression of the anterior cervical pathology was performed successfully via a wider operative field. The symptoms and neurological score improved significantly without any new deficits in all patients except in one with a three-level fusion who needed re-operation for further decompression. No major graft complications such as graft extrusion or pseudoarthrosis occurred. Graft fracture was noted in five cases. However, good bony fusion was observed in all these cases without any further treatment. Normal cervical lordosis was preserved in most cases except in four, who lost lordotic alignment but did not show kyphosis. Major advantages of this method are a wider operative field, excellent graft fusion rate, and no need for an additional incision to obtain autogenous bone graft. These benefits seem have to contributed to satisfactory surgical results in this series.

Osteoplastic anterolateral vertebrotomy without fusion for multilevel cervical ossification of the posterior longitudinal ligament

OBJECTIVE To introduce a new method for osteoplastic anterolateral vertebrotomy without fusion to resect multilevel cervical ossification of the posterior longitudinal ligament. METHODS A multilevel vertebral column graft containing portions of the intervertebral discs is cut with a microsurgical saw from the anterolateral part of the vertebra via a thin lateral gutter placed in advance. A sufficiently wide oblique operative field is provided for resection of the ossified ligament with this method, and only a narrow bony defect remains after simple replacement of the vertebral graft. Postoperatively, patients are allowed to walk, usually within 2 to 3 days, wearing simple cervical collars. RESULTS Twelve patients underwent resection of the ossified ligaments in two to five vertebral bodies. Clinical results were satisfactory except in one patient, who died as a result of a pulmonary embolism that occurred 10 days after surgery. In all of the other patients, the vertebral column grafts were fused within a few months, and there were no graft-related problems. Postoperative lateral radiographs revealed that cervical alignment was preserved, with intervertebral mobility at the operated segment in flexion-extension views. CONCLUSION The osteoplastic anterolateral vertebrotomy method provided a sufficiently wide operative field for satisfactory resection of multilevel cervical ossification of the posterior longitudinal ligament, simplifying graft replacement and postoperative patient care. The cervical alignment was preserved.

Interferon-β delivery via human neural stem cell abates glial scar formation in spinal cord injury.

Glial scar formation is the major impedance to axonal regrowth after spinal cord injury (SCI), and scar-modulating treatments have become a leading therapeutic goal for SCI treatment. In this study, human neural stem cells (NSCs) encoding interferon-β (INF-β) gene were administered intravenously to mice 1 week after SCI. Animals receiving NSCs encoding IFN-β exhibited significant neurobehavioral improvement, electrophysiological recovery, suppressed glial scar formation, and preservation of nerve fibers in lesioned spinal cord. Systemic evaluation of SCI gliosis lesion site with lesion-specific microdissection, genome-wide microarray, and MetaCore pathway analysis identified upregulation of toll-like receptor 4 (TLR4) in SCI gliosis lesion site, and this led us to focus on TLR4 signaling in reactive astrocytes. Examination of primary astrocytes from TLR4 knockout mice, and in vivo inhibition of TLR4, revealed that the effect of IFN-β on the suppression of glial scar formation in SCI requires TLR4 stimulation. These results suggest that IFN-β delivery via intravenous injection of NSCs following SCI inhibits glial scar formation in spinal cord through stimulation of TLR4 signaling.

Treatment of traumatic atlanto-occipital dislocation in chronic phase.

Abstract We report the case of 27-year-old woman who presented with mild neurological deficits with significant anterior dislocation of the atlanto-occipital junction in a chronic phase after initial conservative treatment in another hospital. The importance of early diagnosis and treatment for atlanto-occipital dislocation is emphasized. The dislocation could not be reduced sufficiently either by halo ring cervical traction or surgical procedure 5 months after the accident. Therefore, transoral odontoidectomy for decompression of the medulla, together with the posterior occipitocervical fusion with a titanium loop brace was performed. The patients symptoms disappeared completely within a few months after the operation. Magnetic resonance imaging findings suggesting soft tissue damage is the key to an early diagnosis and subsequent stabilization of traumatic atlanto-occipital dislocation in the early phase. Transoral decompressive odontoidectomy combined with posterior fusion may be considered for the treatment of irreducible atlanto-occipital dislocation in a chronic phase.