Takafumi Mitsuhara

Simulated microgravity facilitates cell migration and neuroprotection after bone marrow stromal cell transplantation in spinal cord injury

IntroductionRecently, cell-based therapy has gained significant attention for the treatment of central nervous system diseases. Although bone marrow stromal cells (BMSCs) are considered to have good engraftment potential, challenges due to in vitro culturing, such as a decline in their functional potency, have been reported. Here, we investigated the efficacy of rat BMSCs (rBMSCs) cultured under simulated microgravity conditions, for transplantation into a rat model of spinal cord injury (SCI).MethodsrBMSCs were cultured under two different conditions: standard gravity (1G) and simulated microgravity attained by using the 3D-clinostat. After 7 days of culture, the rBMSCs were analyzed morphologically, with RT-PCR and immunostaining, and were used for grafting. Adult rats were used for constructing SCI models by using a weight-dropping method and were grouped into three experimental groups for comparison. rBMSCs cultured under 1 g and simulated microgravity were transplanted intravenously immediately after SCI. We evaluated the hindlimb functional improvement for 3 weeks. Tissue repair after SCI was examined by calculating the cavity area ratio and immunohistochemistry.ResultsrBMSCs cultured under simulated microgravity expressed Oct-4 and CXCR4, in contrast to those cultured under 1 g conditions. Therefore, rBMSCs cultured under simulated microgravity were considered to be in an undifferentiated state and thus to possess high migration ability. After transplantation, grafted rBMSCs cultured under microgravity exhibited greater survival at the periphery of the lesion, and the motor functions of the rats that received these grafts improved significantly compared with the rats that received rBMSCs cultured in 1 g. In addition, rBMSCs cultured under microgravity were thought to have greater trophic effects on reestablishment and survival of host spinal neural tissues because cavity formations were reduced, and apoptosis-inhibiting factor expression was high at the periphery of the SCI lesion.ConclusionsHere we show that transplantation of rBMSCs cultured under simulated microgravity facilitates functional recovery from SCI rather than those cultured under 1 g conditions.

Application of 4D-CTA using 320-row area detector computed tomography on spinal arteriovenous fistulae: initial experience

Time-resolved computed tomography angiography (4D-CTA) using a 320-row area detector CT scanner has recently been applied in the evaluation of cranial vascular disorders. However, application of 4D-CTA to spinal vascular disorder evaluation has never before been described. The authors herein report their initial experience of 4D-CTA in the evaluation of spinal arteriovenous fistulas (AVFs) and compare this novel modality with other imaging modalities. Four consecutive patients with spinal AVF underwent time-resolved contrast-enhanced magnetic resonance angiography (trMRA), 4D-CTA, and selective catheter angiography (CA). In 4D-CTA, volume data was transformed into 3D volume-rendered images and maximum intensity projection. These images were also evaluated by time-resolved serial phases. Then, images of each modality were compared, focusing on the detection of perimedullary draining veins and the prediction of AVF location and drainage flow direction. All modalities successfully detected perimedullary draining veins in all cases. Location of the AVF was detected in all cases by CA. trMRA and 4D-CTA detected the AVF in three out of the four cases. With regard to flow direction, while 4D-CTA successfully depicted ascending or descending drainage flow in the spinal canal, CA failed to detect the flow direction in one case while trMRA failed in two cases. In the case with epidural AVF, 4D-CTA was the only technique to detect the flow direction of perimedullary drainage. Although this is only an initial experience of the application of 4D-CTA to spinal vascular diseases, 4D-CTA was capable of detecting the dynamic vascular flow of spinal AVFs. The authors believe that 4D-CTA can be a useful option in the evaluation of spinal AVFs.

Endovascular coil embolization for ruptured kissing aneurysms associated with A1 fenestration.

Background: Fenestration of intracranial arteries is a rare anomaly, and is frequently associated with cerebral aneurysms. In this paper, we report rare kissing aneurysms associated with A1 fenestration. Case Description: A 71-year-old woman presented with subarachnoid hemorrhage. Diagnostic digital subtraction angiography revealed two saccular aneurysms at the proximal junction of a fenestration and posterior aspect of the fenestration that appeared to be ‘kissing’ each other. Emergent endovascular coil embolization was performed. Conclusion: Kissing aneurysms associated with fenestration of the horizontal segment in the anterior cerebral artery are rare, and have not been reported. During treatment of such specific types of aneurysms by endovascular treatment, three-dimensional rotational digital subtraction angiography was very useful for deciding the appropriate working angles.

A Case of a Primary Radiation-Induced Malignant Peripheral Nerve Sheath Tumor in the Cauda Equina in a Patient with NeurofibromatosisType 2

Abstract Background: Radiation-induced malignant peripheral nerve sheath tumors (MPNSTs) may occur in any irradiated area of the body where primary malignant lesions existed. However, there have been isolated reports regarding the radiation-induced transformation of spinal schwannomas into MPNSTs in patients with neurofibromatosis type 2 (NF2). Case material: A 47-year-old woman presented with gradually progressive paraparesis, bowel-bladder dysfunction, and acute consciousness disturbance. Fifteen years previously, she had undergone radical hysterectomy with adjuvant intraoperative para-aortic radiotherapy for the treatment of uterine cervical cancer. Magnetic resonance (MR) images of the brain revealed hydrocephalus and multiple intracranial tumors in bilateral oculomotor and vestibular nerves. Spinal MR images revealed numerous tumors in the cervical cord and cauda equina. Signal alteration in the lumbosacral vertebral bodies on MR images, which implies fatty degeneration, suggested that the cauda equina was included in the previous radiation field. Because the patient complained of severe pain in the lower back region and lower extremities, we partially resected the lumbosacral tumors to achieve pain control. Histological examination of the resected tumors revealed the lesions to be MPNSTs. Despite postoperative radiation therapy, the residual tumor regrew. The patient is now under palliative care. Conclusion: We report a rare case of a primary MPNST in the cauda equina occurring 15 years after the administration of intraoperative irradiation for uterine cancer. In patients with NF2 who undergo irradiation for malignancies, long-term observation is mandatory because tardive malignant transformation may occur in pre-existing schwannomas in the irradiated field.

Three Cases of Moyamoya Disease with a History of Kawasaki Disease

Here, we report three cases of moyamoya disease with a history of Kawasaki disease. A 33-year-old man was found to have stenotic lesions of the internal carotid arteries(ICAs)on both sides at a nearby hospital where he visited complaining of headache and lisping. He had received immunoglobulin therapy for Kawasaki disease at the ages of 1, 2, and 6 years. MRI showed only a chronic ischemic lesion in the white matter. Angiography showed occlusion at the terminal portion of the ICAs on both sides. He was diagnosed with moyamoya disease, but as he had no symptoms and preserved cerebral blood flow (CBF), he was kept under observation. An 8-year-old boy was diagnosed with moyamoya disease and underwent right encephaloduroarteriosynangiosis at a nearby hospital. He had received immunoglobulin therapy for Kawasaki disease at the age of 1 year. His ischemic symptoms worsened. Although MRI detected no apparent ischemic lesion, angiography revealed severe stenosis at the terminal portions of the ICAs on both sides, and 123I-IMP SPECT showed CBF impairment. Bilateral direct bypass was performed. His father was subsequently also diagnosed with moyamoya disease. A 4-year-old girl with epilepsy was diagnosed with moyamoya disease at a nearby hospital. She had been treated with aspirin for Kawasaki disease at the age of 1 year. MRI detected no remarkable ischemic lesions, but angiography revealed mild stenosis at the terminal portions of the ICAs on both sides. Five months later, her ischemic symptoms were worsening with progressing stenotic lesions, and she underwent bilateral direct bypass.

The characteristics of human cranial bone marrow mesenchymal stem cells.

Recently, cell-based therapy has attracted attention for treatment of central nervous system (CNS) disorders. Bone marrow-derived mesenchymal stem cells (BMSCs) are considered to have good engraftment potential. Therefore, more efficient and less invasive methods to obtain donor cells are required. Here, we established human BMSCs from cranial bone waste (cBMSCs) obtained following routine neurosurgical procedures. cBMSCs and cells obtained from the iliac crest (iBMSCs, standard BMSCs) showed expression of cell surface markers associated with mesenchymal stem cells and multipotency traits such as differentiation into osteogenic and adipogenic lineages. cBMSCs showed higher expression of the neural crest-associated mRNAs p75, Slug, and Snail than iBMSCs. Neurogenic induced cells from cBMSCs expressed the neural markers nestin, Pax6, neurofilament (NF)-L, and NF-M as seen with RT-PCR, and NF-M protein as seen with western blotting at higher levels than cells from iBMSCs. Immunostaining showed a significantly greater proportion of NF-M-positive cells in the population of induced cBMSCs compared with the population of iBMSCs. Thus, cBMSCs showed a greater tendency to differentiate into neuron-like cells than iBMSCs.

Gowers' intrasyringeal hemorrhage associated with Chiari type I malformation in Noonan syndrome.

Background: Idiopathic hemorrhage in a syrinx is a rare entity known as Gowers’ intrasyringeal hemorrhage. Bleeding confined to the syrinx cavity causes severe, sometimes acute, neurological deficits. We report a case of intrasyringeal hemorrhage into a preexisting lumbosacral syrinx associated with Chiari type I malformation. Case Description: A 39-year-old female with Noonan syndrome underwent foramen magnum decompression and a cervical syrinx-subarachnoid shunt for Chiari type I malformation-associated syringomyelia 7 years ago. She presented progressive gait deterioration and acute urinary dysfunction, indicating conus medullaris syndrome. Initial magnetic resonance imaging revealed massive hemorrhage in the intrasyringeal cavity of the conus medullaris. The patient underwent surgical removal of the intrasyringeal hematoma and her neurological symptoms improved postoperatively. Conclusion: Although Gowers’ intrasyringeal hemorrhage is rare, this entity should be taken into consideration in patients with syringomyelia showing acute neurological deterioration.

Intraoperative Neurophysiologic Monitoring for Degenerative Cervical Myelopathy

Multimodal intraoperative neurophysiologic monitoring is a reliable tool for detecting intraoperative spine injury and is recommended during surgery for degenerative cervical myopathy (DCM). Somatosensory evoked potential (SEP) can be used to monitor spine and peripheral nerve injury during positioning in surgery for DCM. Compensation technique for transcranial evoked muscle action potentials (tcMEPs) should be adopted in intraoperative monitoring during surgery for DCM. Free-running electromyography is a useful real-time monitoring add-on modality in addition to SEP and tcMEP.

Factors Related to Frailty Associated with Clinical Deterioration After Meningioma Surgery in the Elderly

BACKGROUND Older patients are increasingly presenting for surgery with intracranial meningioma because of progress with diagnostic imaging and longer life expectancy. However, older patients have many problems, such as comorbidities and reduced physiological capacity reflected in the frailty index. This study examines the factors affecting clinical deterioration after surgery in older patients, particularly factors associated with frailty. METHODS Two hundred sixty-five patients older than 65 years underwent surgical resection of meningioma at Hiroshima University and related hospitals between 2000 and 2016. Karnofsky Performance Status (KPS) scores before and after surgery were evaluated. Factors related to the deterioration of KPS were analyzed with multivariate logistic regression modeling, including body mass index and serum albumin. RESULTS KPS score deteriorated compared with preoperative score in 56 patients at discharge and in 40 patients at 3 months later, and 2 patients died within 1 year after surgery. Multivariate logistic regression analysis in addition to preoperative body mass index and serum albumin indicated skull base tumor location (odds ratio [OR], 4.67; 95% confidence interval [CI], 2.02-10.8) and serum albumin (OR, 2.38; 95% CI, 1.06-5.34) were risk factors for deterioration of KPS score at discharge. Age (OR, 0.91; 95% CI, 0.85-0.98), skull base tumor location (OR, 4.32; 95% CI, 1.45-12.9), tumor size (OR, 1.03; 95% CI, 1.00-1.05), and serum albumin (OR, 3.53; 95% CI, 1.29-9.61) were significant risk factors for perioperative intracranial complications. CONCLUSIONS Skull base tumor location and serum albumin correlated with deterioration of clinical status after surgery.

Rat Cranial Bone-Derived Mesenchymal Stem Cell Transplantation Promotes Functional Recovery in Ischemic Stroke Model Rats

The functional disorders caused by central nervous system (CNS) diseases, such as ischemic stroke, are clinically incurable and current treatments have limited effects. Previous studies suggested that cell-based therapy using mesenchymal stem cells (MSCs) exerts therapeutic effects for ischemic stroke. In addition, the characteristics of MSCs may depend on their sources. Among the derived tissues of MSCs, we have focused on cranial bones originating from the neural crest. We previously demonstrated that the neurogenic potential of human cranial bone-derived MSCs (cMSCs) was higher than that of human iliac bone-derived MSCs. Therefore, we presumed that cMSCs have a higher therapeutic potential for CNS diseases. However, the therapeutic effects of cMSCs have not yet been elucidated in detail. In the present study, we aimed to demonstrate the therapeutic effects of transplantation with rat cranial bone-derived MSCs (rcMSCs) in ischemic stroke model rats. The mRNA expression of brain-derived neurotrophic factor and nerve growth factor was significantly stronger in rcMSCs than in rat bone marrow-derived MSCs (rbMSCs). Ischemic stroke model rats in the rcMSC transplantation group showed better functional recovery than those in the no transplantation and rbMSC transplantation groups. Furthermore, in the in vitro study, the conditioned medium of rcMSCs significantly suppressed the death of neuroblastoma × glioma hybrid cells (NG108-15) exposed to oxidative and inflammatory stresses. These results suggest that cMSCs have potential as a candidate cell-based therapy for CNS diseases.