Koshi Ninomiya

Intranasal delivery of bone marrow stromal cells to spinal cord lesions

OBJECT The intranasal delivery of bone marrow stromal cells (BMSCs) or mesenchymal stem cells to the injured brains of rodents has been previously reported. In this study, the authors investigated whether BMSCs migrate to spinal cord lesions through an intranasal route and whether the administration affected functional recovery. METHODS Forty Sprague-Dawley rats that were subjected to spinal cord injuries at the T7-8 level were divided into 5 groups (injured + intranasal BMSC-treated group, injured + intrathecal BMSC-treated group, injured-only group, injured + intranasal vehicle-treated group, and injured + intrathecal vehicle-treated group). The Basso-Beattie-Bresnahan (BBB) scale was used to assess hind limb motor functional recovery for 2 or 4 weeks. Intralesionally migrated BMSCs were examined histologically and counted at 2 and 4 weeks. To evaluate the neuroprotective and trophic effects of BMSCs, the relative volume of the lesion cavity was measured at 4 weeks. In addition, nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) levels in the CSF were evaluated at 2 weeks. RESULTS Intranasally administered BMSCs were confirmed within spinal cord sections at both 2 and 4 weeks. The highest number, which was detected in the intrathecal BMSC-treated group at 2 weeks, was significantly higher than that in all the other groups. The BBB score of the intranasal BMSC-treated group showed statistically significant improvements by 1 week compared with the control group. However, in the final BBB scores, there was a statistically significant difference only between the intrathecal BMSC-treated group and the control group. The cavity ratios in the BMSC-treated groups were smaller than those of the control groups, but the authors did not find any significant differences in the NGF and BDNF levels in the CSF among the treatment and control groups. CONCLUSIONS BMSCs reached the injured spinal cord through the intranasal route and contributed to the recovery of hind limb motor function and lesion cavity reduction. However, the effects were not as significant as those seen in the intrathecal BMSC-treated group.

Isolation of human adult olfactory sphere cells as a cell source of neural progenitors

Olfactory stem cells are generated from olfactory mucosa. Various culture conditions generate olfactory stem cells that differ according to species and developmental stage and have different progenitor or stem cell characteristics. Olfactory spheres (OSs) are clusters of progenitor or stem cells generated from olfactory mucosa in suspension culture. In this study, adult human OSs were generated and their characteristics analyzed. Human OSs were adequately produced from olfactory mucosa with area over 40 mm(2). Immunocytochemistry (ICC) and fluorescence-activated cell sorting showed that human OSs were AN2 and A2B5-positive. Immunofluorescence analysis of cell type-specific ICC indicated that the number of Tuj1-positive OS cells was significantly elevated. Tuj1-positive cells displayed typical neuronal soma and dendritic morphology. Human OS cells were also immunopositive for MAP2. By contrast, few RIP-, O4-, and GFAP-positive cells were present. These RIP, O4, and GFAP-positive cells did not resemble bona fide oligodendrocytes and astrocytes morphologically. In culture to induce differentiation of oligodendrocytes, human OS cells also expressed neuronal markers, but neither oligodendrocyte or astrocyte markers. These findings suggest that human OS cells autonomously differentiate into neurons in our culture condition and have potential to be used as a cell source of neural progenitors for their own regenerative grafts, avoiding the need for immunosuppression and ethical controversies.

Presence of trans-synaptic neurons derived from olfactory mucosa transplanted after spinal cord injury.

Study Design. Using biotinylated dextran amine (BDA) and wheat germ agglutinin (WGA) tracers, we measured the effectiveness of olfactory mucosa (OM) transplantation as a scaffold in a rat model of chronic spinal cord injury (SCI). Objective. We examined whether OM transplantation for chronic SCI in rats results in reconstruction of neuronal pathways by both regeneration of the remaining axons and supply of OM-derived trans-synaptic neurons. Summary of Background Data. OM is one of the ideal scaffolds for axonal regeneration after chronic SCI. Methods. Rats received a mild contusion at vertebral level T6–T7. Two weeks after SCI, enhanced green fluorescent protein rat-derived OM, respiratory mucosa, and phosphate-buffered saline were transplanted into each group of SCI rats. Ten weeks after SCI, BDA was injected into the right sensorimotor cortex. Eleven weeks after SCI, WGA was injected into the L1–L2 posterior column to label the corticospinal tract retrogradely and trans-synaptically. Twelve weeks after SCI, rats were killed and their spinal cords were divided into cervical (area a), thoracic-injured (area b), and lower thoracic portions (area c). Immunohistochemically, sections of area (b) were evaluated by counting cells positive for enhanced green fluorescent protein, 4′,6-diamidino-2-phenylindole, WGA, and BDA (OM and respiratory mucosa groups). Axonal regenerations were estimated by counting WGA- and BDA-positive dots in transverse sections of area (a) and area (c). Results. Compared with respiratory mucosa and phosphate-buffered saline transplantation, OM transplantation increased the number of WGA-positive dots in area (a), and the number of BDA-positive dots in area (c) was more after OM transplantation than after phosphate-buffered saline transplantation. Furthermore, the number of quadruple-positive cells in area (b) was much higher after OM transplantation. Conclusion. Our results provide both indirect and direct evidence for the presence of trans-synaptic neurons. OM transplantation in rats with chronic SCI resulted in reconstruction of neural pathways by both providing trans-synaptic neurons and supporting regeneration of remaining axons. The olfactory mucosa is thought to be an efficacious scaffold to produce the relay neuron in chronic spinal cord injury. Level of Evidence: N/A

Radiological Evaluation of the Initial Fixation between Cortical Bone Trajectory and Conventional Pedicle Screw Technique for Lumbar Degenerative Spondylolisthesis

Study Design Retrospective study. Purpose To compare initial fixation using the cortical bone trajectory (CBT) technique versus conventional pedicle screws (PS) in radiographs of postsurgical lumbar degenerative spondylolisthesis. Overview of Literature Few reports have documented the holding strength of CBT technique for spondylolisthesis cases. Methods From October 2009 to June 2014, 21 cases of degenerative spondylolisthesis were surgically treated in our institution. Ten were treated with conventional PS technique and 11 of with CBT technique. Mean lumbar lordosis and percent slippage were evaluated preoperatively, immediately after surgery, and 6 months and 1 year postoperatively using radiographs. We also investigated percent loss of slip reduction. Results There were statistically significant differences between preoperative percent slippage and postoperative slippage in both PS and CBT procedures over 1 year, and both techniques showed good slip reduction. On the other hand, lumbar lordosis did not change significantly in either the PS or CBT groups over 1 year. Conclusions CBT technique showed similarly good initial fixation compared with the PS procedure in the treatment of lumbar degenerative spondylolisthesis.

Clear Zone Formation around Screws in the Early Postoperative Stages after Posterior Lumbar Fusion Using the Cortical Bone Trajectory Technique

Study Design Retrospective study. Purpose To evaluate the initial fixation using the cortical bone trajectory (CBT) technique for posterior lumbar fusion through assessment of the clear zones around the screws and the risk factors involved. Overview of Literature Postoperative radiolucent zones (clear zones) are an indicator of poor conventional pedicle screw fixation. Methods Between January 2013 and April 2014, 19 patients (8 men and 11 women) underwent posterior lumbar interbody fusion or posterior lumbar fusion using the CBT technique. A total of 109 screws were used for evaluation with measurement of the maximum insertional torque of last two screw rotations. Clear zone-positivity on plain radiographs was investigated 6 months after surgery. The relation between intraoperative insertional torque and clear zone-positivity was investigated by one-way analysis of variance. In addition, the correlation between clear zone-positivity and gender, age (<75 years old or >75 years old), or operative stabilization level (<2 or >3 vertebral levels) was evaluated using the chi-square test. Results Clear zones were observed around six screws (5.50%) in five patients (26.3%). The mean insertional torque (4.00±2.09 inlbs) of clear zone-positive screws was lower than that of clear zone-negative screws (8.12±0.50 in-lbs), but the difference was not significant. There was a significant correlation between clear zone-positivity and operative level of stabilization. Conclusions The low incidence of clear zone-positive screws indicates good initial fixation using the CBT technique. Multilevel fusions may be risk factors for clear zone generation.

Significance of the Pars Interarticularis in the Cortical Bone Trajectory Screw Technique: An In Vivo Insertional Torque Study

Study Design Retrospective study. Purpose Cortical bone trajectory (CBT), a more medial-to-lateral and shorter path than the traditional one for spinal fusion, is thought to be effective for severely degenerated vertebrae because screws are primarily stabilized at the posterior elements. We evaluated the efficacy of this approach through in vivo insertional torque measurement. Overview of Literature There has been only one prior in vivo study on CBT insertional torque. Methods Between January 2013 and April 2014, a total of 22 patients underwent posterior lumbar fusion using the CBT technique. The maximum insertional torque, which covers the radial strength needed for insertion, was measured for 113 screws, 8 of which were inserted for L5 spondylolysis. The insertional torque for cases with (n=8) and without (n=31) spondylolysis of L5 were compared using one-way analysis of variance (ANOVA). To evaluate vertebral degeneration, we classified 53 vertebrae without spondylolysis by lumbar radiography using semiquantitative methods; the insertional torque for the 105 screws used was compared on the basis of this classification. Additionally, differences in insertional torque among cases grouped by age, sex, and lumbar level were evaluated for these 105 screws using ANOVA and the Tukey test. Results The mean insertional torque was significantly lower for patients with spondylolysis than for those without spondylolysis (4.25 vs. 8.24 in-lb). There were no statistical differences in insertional torque according to vertebral grading or level. The only significant difference in insertional torque between age and sex groups was in men <75 years and women ≥75 years (10 vs. 5.5 in-lb). Conclusions Although CBT should be used with great caution in patient with lysis who are ≥75 years, it is well suited for dealing with severely degenerated vertebrae because the pars interarticularis plays a very important role in the implementation of this technique.

Differences between Cervical Schwannomas of the Anterior and Posterior Nerve Roots in Relation to the Incidence of Postoperative Radicular Dysfunction.

Study Design A retrospective study. Purpose To assess the case files of patients who underwent surgery for cervical dumbbell schwannoma for determining the differences between schwannomas of the anterior and posterior nerve roots with respect to the incidence of postoperative radicular dysfunction. Overview of Literature The spinal roots giving origin to schwannoma are frequently nonfunctional, but there is a risk of postoperative neurological deficit once these roots are resected during surgery. Methods Fifteen patients with cervical dumbbell schwannomas were treated surgically. Ten men and 5 women, who were 35-79 years old (mean age, 61.5 years), presented with neck pain (n=6), radiculopathy (n=10), and myelopathy (n=11). Results Fourteen patients underwent gross total resection and exhibited no recurrence. Follow-ups were performed for a period of 6-66 months (mean, 28 months). Preoperative symptoms resolved in 11 patients (73.3%) but they persisted partially in 4 patients (26.7%). Six patients had tumors of anterior nerve root origin, and 9 patients had tumors of posterior nerve root origin. Two patients who underwent total resection of anterior nerve root tumors (33.3%) displayed minor postoperative motor weakness. One patient who underwent total resection of a posterior nerve root tumor (11.1%) showed postoperative numbness. Conclusions Appropriate tumor removal improved the neurological symptoms. In this study, the incidence of radicular dysfunction was higher in patients who underwent resection of anterior nerve root tumors than in patients who underwent resection of posterior nerve root tumors.

Syringomyelia associated with spinal arachnoiditis treated by partial arachnoid dissection and syrinx-far distal subarachnoid shunt.

The authors describe a new modified surgical approach to minimize the postoperative recurrence of a syrinx after surgery to treat syringomyelia associated with spinal adhesive arachnoiditis in two cases. Both patients presented with progressive gait disturbance without any remarkable history, and spinal magnetic resonance imaging revealed a syrinx and broad irregular disappearance of the subarachnoid space and/or deformity of the cord. We successfully performed a partial arachnoid dissection and syrinx-far distal subarachnoid shunt for both cases.

Isthmus-guided Cortical Bone Trajectory Reduces Postoperative Increases in Serum Creatinine Phosphokinase Concentrations

This study aimed to determine whether an isthmus‐guided cortical bone trajectory (CBT) technique provides better clinical outcomes than the original cortical bone trajectory CBT technique for screw fixation.

A Pilot Clinical Study of Olfactory Mucosa Autograft for Chronic Complete Spinal Cord Injury

Recent studies of spinal cord axon regeneration have reported good long-term results using various types of tissue scaffolds. Olfactory tissue allows autologous transplantation and can easily be obtained by a simple biopsy that is performed through the external nares. We performed a clinical pilot study of olfactory mucosa autograft (OMA) for chronic complete spinal cord injury in eight patients according to the procedure outlined by Lima et al. Our results showed no serious adverse events and improvement in both the American Spinal Injury Association (ASIA) Impairment Scale (AIS) grade and ASIA motor score in five patients. The preoperative post-rehabilitation ASIA motor score improved from 50 in all cases to 52 in case 2, 60 in case 4, 52 in case 6, 55 in case 7, and 58 in case 8 at 96 weeks after OMA. The AIS improved from A to C in four cases and from B to C in one case. Motor evoked potentials (MEPs) were also seen in one patient, reflecting conductivity in the central nervous system, including the corticospinal tract. The MEPs induced with transcranial magnetic stimulation allow objective assessment of the integrity of the motor circuitry comprising both the corticospinal tract and the peripheral motor nerves.We show the feasibility of OMA for chronic complete spinal cord injury.