Updated on 2025/08/22

写真a

 
UKAI Ryou
 
Organization
Institute of Regenerative Medicine Department of Neural Regenerative Medicine Lecturer
Title
Lecturer
ORCID ID
0000-0002-8391-2552
External link

Research Areas

  • Life Science / Neurosurgery

Education

  • Sapporo Medical University   Graduate School of Medicine

    2002.4 - 2006.3

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  • Sapporo Medical University

    1995.4 - 2001.3

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Research History

  • Sapporo Medical University   Research Institute for Frontier Medicine   Lecturer

    2023.4

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Papers

  • Intravenous Infusion of Autologous Mesenchymal Stem Cells Expanded in Auto Serum for Chronic Spinal Cord Injury Patients: A Case Series. International journal

    Ryosuke Hirota, Masanori Sasaki, Satoshi Iyama, Kota Kurihara, Ryunosuke Fukushi, Hisashi Obara, Tsutomu Oshigiri, Tomonori Morita, Masahito Nakazaki, Takahiro Namioka, Ai Namioka, Rie Onodera, Yuko Kataoka-Sasaki, Shinichi Oka, Mitsuhiro Takemura, Ryo Ukai, Takahiro Yokoyama, Yuichi Sasaki, Tatsuro Yamashita, Masato Kobayashi, Yusuke Okuma, Reiko Kondo, Ryo Aichi, Satoko Ohmatsu, Noritaka Kawashima, Yoichi M Ito, Masayoshi Kobune, Kohichi Takada, Sumio Ishiai, Toru Ogata, Atsushi Teramoto, Toshihiko Yamashita, Jeffery D Kocsis, Osamu Honmou

    Journal of clinical medicine   13 ( 20 )   2024.10

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    Objective: The safety, feasibility, and potential functional improvement following the intravenous infusion of mesenchymal stem cells (MSCs) were investigated in patients with chronic severe spinal cord injury (SCI). Methods: The intravenous infusion of autologous MSCs cultured in auto-serum under Good Manufacturing Practices (GMP) was administered to seven patients with chronic SCI (ranging from 1.3 years to 27 years after the onset of SCI). In addition to evaluating feasibility and safety, neurological function was evaluated using the American Spinal Injury Association Impairment Scale (AIS), International Standards for Neurological Classification of Spinal Cord Injury (ISCSCI-92), and Spinal Cord Independence Measure III (SCIM-III). Results: No serious adverse events occurred. Neither CNS tumors, abnormal cell growth, nor neurological deterioration occurred in any patients. While this initial case series was not blinded, significant functional improvements and increased quality of life (QOL) were observed at 90 and 180 days post-MSC infusion compared to pre-infusion status. One patient who had an AIS grade C improved to grade D within six months after MSC infusion. Conclusions: This case series suggests that the intravenous infusion of autologous MSCs is a safe and feasible therapeutic approach for chronic SCI patients. Furthermore, our data showed significant functional improvements and better QOL after MSC infusion in patients with chronic SCI. A blind large-scale study will be necessary to fully evaluate this possibility.

    DOI: 10.3390/jcm13206072

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  • Intravenous Infusion of Autologous Mesenchymal Stem Cells Expanded in Auto Serum for Chronic Spinal Cord Injury Patients: A Case Series

    Ryosuke Hirota, Masanori Sasaki, Satoshi Iyama, Kota Kurihara, Ryunosuke Fukushi, Hisashi Obara, Tsutomu Oshigiri, Tomonori Morita, Masahito Nakazaki, Takahiro Namioka, Ai Namioka, Rie Onodera, Yuko Kataoka-Sasaki, Shinichi Oka, Mitsuhiro Takemura, Ryo Ukai, Takahiro Yokoyama, Yuichi Sasaki, Tatsuro Yamashita, Masato Kobayashi, Yusuke Okuma, Reiko Kondo, Ryo Aichi, Satoko Ohmatsu, Noritaka Kawashima, Yoichi M. Ito, Masayoshi Kobune, Kohichi Takada, Sumio Ishiai, Toru Ogata, Atsushi Teramoto, Toshihiko Yamashita, Jeffery D. Kocsis, Osamu Honmou

    Journal of Clinical Medicine   13 ( 20 )   2024.10

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    DOI: 10.3390/jcm13206072

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  • 慢性期重症頭部外傷症例に対する自家骨髄MSC静脈投与

    岡 真一, 八巻 智洋, 佐々木 祐典, 佐々木 優子, 浪岡 隆洋, 浪岡 愛, 中崎 公仁, 竹村 光広, 鵜飼 亮, 横山 貴裕, 小瀧 勝, 岩立 康男, 小林 繁樹, 本望 修

    Journal of Japan Coma Society: JJCS   32 ( 1 )   103 - 103   2024.7

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    Language:Japanese   Publisher:(一社)日本意識障害学会  

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  • Intravenous infusion of auto-serum-expanded autologous mesenchymal stem cells into chronic severe brain injury patients

    Tomohiro Yamaki, Shinichi Oka, Satoshi Iyama, Masanori Sasaki, Rie Onodera, Yuko Kataoka-Sasaki, Takahiro Namioka, Ai Namioka, Masahito Nakazaki, Mitsuhiro Takemura, Ryo Ukai, Takahiro Yokoyama, Yuichi Sasaki, Tatsuro Yamashita, Masato Kobayashi, Misako Yamaguchi, Marina Fukino, Taro Takazawa, Megumi Hayasaka, Takamitsu Owaku, Mika Funakura, Shinji Onodera, Yoichi M. Ito, Masayoshi Kobune, Junji Kato, Sumio Ishiai, Jeffery D. Kocsis, Masaru Odaki, Yasuo Iwadate, Shigeki Kobayashi, Osamu Honmou

    Interdisciplinary Neurosurgery   36   101927 - 101927   2024.6

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    Publishing type:Research paper (scientific journal)   Publisher:Elsevier BV  

    DOI: 10.1016/j.inat.2023.101927

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  • Rehabilitation facilitates functional improvement following intravenous infusion of mesenchymal stem cells in the chronic phase of cerebral ischemia in rats. International journal

    Tatsuro Yamashita, Masanori Sasaki, Yuichi Sasaki, Hiroshi Nagahama, Shinichi Oka, Yuko Kataoka-Sasaki, Ryo Ukai, Takahiro Yokoyama, Masato Kobayashi, Masafumi Kakizawa, Jeffery D Kocsis, Osamu Honmou

    Brain research   1825   148709 - 148709   2024.2

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    The primary objective of this study was to investigate the potential facilitating effects of daily rehabilitation for chronic cerebral ischemia following the intravenous infusion of mesenchymal stem cells (MSC) in rats. The middle cerebral artery (MCA) was occluded by intraluminal occlusion using a microfilament (MCAO). Eight weeks after MCAO induction, the rats were used as a chronic cerebral ischemia model. Four experimental groups were studied: Vehicle group (medium only, no cells); Rehab group (vehicle + rehabilitation), MSC group (MSC only); and Combined group (MSC + rehabilitation). Rat MSCs were intravenously infused eight weeks after MCAO induction, and the rats received daily rehabilitation through treadmill exercise for 20 min. Behavioral testing, lesion volume assessment using magnetic resonance imaging (MRI), and histological analysis were performed during the observation period until 16 weeks after MCAO induction. All treated animals showed functional improvement compared with the Vehicle group; however, the therapeutic efficacy was greatest in the Combined group. The combination therapy is associated with enhanced neural plasticity shown with histological analysis and MRI diffusion tensor imaging. These findings provide behavioral evidence for enhanced recovery by combined therapy with rehabilitation and intravenous infusion of MSCs, and may form the basis for the development of clinical protocols in the future.

    DOI: 10.1016/j.brainres.2023.148709

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  • Therapeutic efficacy of intravenous infusion of mesenchymal stem cells in rat perinatal brain injury. International journal

    Kojiro Terada, Masanori Sasaki, Hiroshi Nagahama, Yuko Kataoka-Sasaki, Shinichi Oka, Ryo Ukai, Takahiro Yokoyama, Yusuke Iizuka, Takuro Sakai, Shinobu Fukumura, Takeshi Tsugawa, Jeffery D Kocsis, Osamu Honmou

    Pediatric research   2023.7

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    BACKGROUND: Perinatal brain injury is multifactorial and primarily associated with brain prematurity, inflammation, and hypoxia-ischemia. Although recent advances in perinatal medicine have improved the survival rates of preterm infants, neurodevelopmental disorders remain a significant complication. We tested whether the intravenous infusion of mesenchymal stem cells (MSCs) had therapeutic efficacy against perinatal brain injury in rats. METHODS: Pregnant rats at embryonic day (E) 18 received lipopolysaccharide and the pups were born at E21. On postnatal day (PND) 7, the left common carotid artery of each pup was ligated, and they were exposed to 8% oxygen for 2 h. They were randomized on PND10, and MSCs or vehicle were intravenously infused. We performed behavioral assessments, measured brain volume using MRI, and performed histological analyses on PND49. RESULTS: Infused MSCs showed functional improvements in our model. In vivo MRI revealed that MSC infusion increased non-ischemic brain volume compared to the vehicle group. Histological analyses showed that cortical thickness, the number of NeuN+ and GAD67+ cells, and synaptophysin density in the non-ischemic hemisphere in the MSC group were greater than the vehicle group, but less than the control group. CONCLUSIONS: Infused MSCs improve sensorimotor and cognitive functions in perinatal brain injury and enhance neuronal growth. IMPACT: Intravenous infusion of MSCs improved neurological function in rats with perinatal brain injury, including motor, sensorimotor, cognitive, spatial, and learning memory. Infused MSCs increased residual (non-ischemic) tissue volume, number of neuronal cells, GABAergic cells, and cortical synapses in the contralesional (right) hemisphere. Intravenous administration of MSC might be suitable for the treatment of perinatal brain injury.

    DOI: 10.1038/s41390-023-02717-9

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  • A practical protocol for high-spatial-resolution magnetic resonance angiography for cerebral arteries in rats. International journal

    Hiroshi Nagahama, Masanori Sasaki, Katsuya Komatsu, Kaori Sato, Yoshimi Katagiri, Masaki Kamagata, Yuko Kataoka-Sasaki, Shinichi Oka, Ryo Ukai, Takahiro Yokoyama, Kojiro Terada, Masato Kobayashi, Jeffery D Kocsis, Osamu Honmou

    Journal of neuroscience methods   386   109784 - 109784   2023.1

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    BACKGROUND: Magnetic resonance angiography (MRA) is an important tool in rat models of cerebrovascular disease. Although MRA has long been used in rodents, the image quality is typically not as high as that observed in clinical practice. Moreover, studies on MRA image quality in rats are limited. This study aimed to develop a practical high-spatial-resolution MRA protocol for imaging cerebral arteries in rats. NEW METHOD: We used the "half position method" regarding coil placement and modified the imaging parameters and image reconstruction method. We applied this new imaging method to measure maturation-related signal changes on rat MRAs. RESULTS: The new practical high-spatial-resolution MRA imaging protocol obtained a signal intensity up to 3.5 times that obtained using a basic coil system, simply by modifying the coil placement method. This method allowed the detection of a gradual decrease in the signal in cerebral vessels with maturation. COMPARISON WITH EXISTING METHODS: A high-spatial-resolution MRA for rats was obtained with an imaging time of approximately 100 min. Comparable resolution and image quality were obtained using the new protocol with an imaging time of 30 min CONCLUSIONS: The new practical high-spatial-resolution MRA protocol can be implemented simply and successfully to achieve high image quality with an imaging time of approximately 30 min. This protocol will benefit researchers performing MRA imaging in cerebral artery studies in rats.

    DOI: 10.1016/j.jneumeth.2023.109784

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  • Enhanced Network in Corticospinal Tracts after Infused Mesenchymal Stem Cells in Spinal Cord Injury. International journal

    Ryosuke Hirota, Masanori Sasaki, Yuko Kataoka-Sasaki, Tsutomu Oshigiri, Kota Kurihara, Ryunosuke Fukushi, Shinichi Oka, Ryo Ukai, Mitsunori Yoshimoto, Jeffery D Kocsis, Toshihiko Yamashita, Osamu Honmou

    Journal of neurotrauma   39 ( 23-24 )   1665 - 1677   2022.12

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    Although limited spontaneous recovery occurs after spinal cord injury (SCI), current knowledge reveals that multiple forms of axon growth in spared axons can lead to circuit reorganization and a detour or relay pathways. This hypothesis has been derived mainly from studies of the corticospinal tract (CST), which is the primary descending motor pathway in mammals. The major CST is the dorsal CST (dCST), being the major projection from cortex to spinal cord. Two other components often called "minor" pathways are the ventral and the dorsal lateral CSTs, which may play an important role in spontaneous recovery. Intravenous infusion of mesenchymal stem cells (MSCs) provides functional improvement after SCI with an enhancement of axonal sprouting of CSTs. Detailed morphological changes of CST pathways, however, have not been fully elucidated. The primary objective was to evaluate detailed changes in descending CST projections in SCI after MSC infusion. The MSCs were infused intravenously one day after SCI. A combination of adeno-associated viral vector (AAV), which is an anterograde and non-transsynaptic axonal tracer, was injected 14 days after SCI induction. The AAV with advanced tissue clearing techniques were used to visualize the distribution pattern and high-resolution features of the individual axons coursing from above to below the lesion. The results demonstrated increased observable axonal connections between the dCST and axons in the lateral funiculus, both rostral and caudal to the lesion core, and an increase in observable axons in the dCST below the lesion. This increased axonal network could contribute to functional recovery by providing greater input to the spinal cord below the lesion.

    DOI: 10.1089/neu.2022.0106

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  • Intravenous Infusion of Autoserum-Expanded Autologous Mesenchymal Stem Cells in Patients With Chronic Brain Injury: Protocol for a Phase 2 Trial. International journal

    Shinichi Oka, Tomohiro Yamaki, Masanori Sasaki, Ryo Ukai, Mitsuhiro Takemura, Takahiro Yokoyama, Yuko Kataoka-Sasaki, Rie Onodera, Yoichi M Ito, Shigeki Kobayashi, Jeffery D Kocsis, Yasuo Iwadate, Osamu Honmou

    JMIR research protocols   11 ( 7 )   e37898   2022.7

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    BACKGROUND: Brain injuries resulting from motor vehicle accidents and falls, as well as hypoxic insults and other conditions, are one of the leading causes of disability and death in the world. Current treatments are limited but include continuous rehabilitation, especially for chronic brain injury. Recent studies have demonstrated that the intravenous infusion of mesenchymal stem cells (MSCs) has therapeutic efficacy for several neurological diseases, including stroke and spinal cord injury. OBJECTIVE: The objective of our investigator-initiated clinical trial is to assess the safety and potential efficacy of the intravenous infusion of autoserum-expanded autologous MSCs for patients with chronic brain injury. METHODS: The (phase 2) trial will be a single-arm, open-label trial with the primary objective of confirming the safety and efficacy of autoserum-expanded autologous MSCs (STR-01; produced under good manufacturing practices) when administered to patients with chronic brain injury. The estimated number of enrolled participants is 6 to 20 patients with a modified Rankin Scale grade of 3 to 5. The assessment of safety and the proportion of cases in which the modified Rankin Scale grade improves by 1 point or more at 180 days after the injection of STR-01 will be performed after MSC infusion. RESULTS: We received approval for our clinical trial from the Japanese Pharmaceuticals and Medical Devices Agency on December 12, 2017. The trial will be completed on June 11, 2023. The registration term is 5 years. The recruitment of the patients for this trial started on April 20, 2018, at Sapporo Medical University Hospital in Japan. CONCLUSIONS: Our phase 2 study will aim to address the safety and efficacy of the intravenous infusion of MSCs for patients with chronic brain injury. The use of STR-01 has been performed for patients with cerebral infarction and spinal cord injury, providing encouraging results. The potential therapeutic efficacy of the systemic administration of autoserum-expanded autologous MSCs for chronic brain injury should be evaluated, given its safety and promising results for stroke and spinal cord injury. TRIAL REGISTRATION: Japan Medical Association Center for Clinical Trials JMA-IIA00333; https://tinyurl.com/nzkdfnbc. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/37898.

    DOI: 10.2196/37898

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  • Pharmacological Difference Between Platelet Aggregations in Cardioembolic Stroke Patients with Direct Oral Anticoagulants: A Pilot Study. International journal

    Masahito Nakazaki, Shinichi Oka, Hirotoshi Magota, Ryo Kiyose, Rie Onodera, Ryo Ukai, Yuko Kataoka-Sasaki, Masanori Sasaki, Osamu Honmou

    Journal of stroke and cerebrovascular diseases : the official journal of National Stroke Association   31 ( 7 )   106520 - 106520   2022.7

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    Background Selecting the appropriate direct oral anticoagulants (DOACs) for embolic ischemic stroke patients, especially on concurrent antiplatelet therapy, is important. However, a limited number of studies have reported on the pharmacological differences in platelet aggregation of each DOAC. We aimed to evaluate the antiplatelet effects of selected DOACs, by comparing dabigatran (a direct oral thrombin inhibitor) and factor Xa (FXa) inhibitors (apixaban and rivaroxaban) in patients who had suffered a cardioembolic stroke. Methods We retrospectively evaluated 12 patients diagnosed with a cardioembolic stroke who took any DOAC without an antiplatelet drug and underwent platelet aggregation tests within 60 days from the onset of symptoms. The platelet aggregation tests were analyzed by both light transmission aggregometry and VerifyNow®. Results Six patients (50%) took dabigatran, while the other six (50%) took an FXa inhibitor (n = 4 for apixaban and n = 2 for rivaroxaban). From the light transmission aggregometry analysis, it was found that the maximal extent of aggregation for adenosine diphosphate (ADP) was significantly higher with dabigatran than with FXa inhibitors, and the ED50 value of ADP on platelet aggregation was significantly lower with dabigatran than with FXa inhibitors. Moreover, the VerifyNow® analyses revealed that P2Y12 reaction units were significantly higher with dabigatran than with FXa inhibitors. Conclusions Dabigatran had little impact on platelet aggregation compared to FXa inhibitors in patients who had suffered a cardioembolic stroke with atrial fibrillation, and who took DOACs for secondary prevention within 60 days from the onset.

    DOI: 10.1016/j.jstrokecerebrovasdis.2022.106520

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  • Repeated intravenous infusion of mesenchymal stem cells for enhanced functional recovery in a rat model of chronic cerebral ischemia. International journal

    Mitsuhiro Takemura, Masanori Sasaki, Yuko Kataoka-Sasaki, Ryo Kiyose, Hiroshi Nagahama, Shinichi Oka, Ryo Ukai, Takahiro Yokoyama, Jeffery D Kocsis, Tetsuya Ueba, Osamu Honmou

    Journal of neurosurgery   1 - 10   2021.12

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    OBJECTIVE: Stroke is a major cause of long-term disability, and there are few effective treatments that improve function in patients during the chronic phase of stroke. Previous research has shown that single systemic infusion of mesenchymal stem cells (MSCs) improves motor function in acute and chronic cerebral ischemia models in rats. A possible mechanism that could explain such an event includes the enhanced neural connections between cerebral hemispheres that contribute to therapeutic effects. In the present study, repeated infusions (3 times at weekly intervals) of MSCs were administered in a rat model of chronic stroke to determine if multiple dosing facilitated plasticity in neural connections. METHODS: The authors induced middle cerebral artery occlusion (MCAO) in rats and, 8 weeks thereafter, used them as a chronic stroke model. The rats with MCAO were randomized and intravenously infused with vehicle only (vehicle group); with MSCs at week 8 (single administration: MSC-1 group); or with MSCs at weeks 8, 9, and 10 (3 times, repeated administration: MSC-3 group) via femoral veins. Ischemic lesion volume and behavioral performance were examined. Fifteen weeks after induction of MCAO, the thickness of the corpus callosum (CC) was determined using Nissl staining. Immunohistochemical analysis of the CC was performed using anti-neurofilament antibody. Interhemispheric connections through the CC were assessed ex vivo by diffusion tensor imaging. RESULTS: Motor recovery was better in the MSC-3 group than in the MSC-1 group. In each group, there was no change in the ischemic volume before and after infusion. However, both thickness and optical density of neurofilament staining in the CC were greater in the MSC-3 group, followed by the MSC-1 group, and then the vehicle group. The increased thickness and optical density of neurofilament in the CC correlated with motor function at 15 weeks following induction of MCAO. Preserved neural tracts that ran through interhemispheric connections via the CC were also more extensive in the MSC-3 group, followed by the MSC-1 group and then the vehicle group, as observed ex vivo using diffusion tensor imaging. CONCLUSIONS: These results indicate that repeated systemic administration of MSCs over 3 weeks resulted in greater functional improvement as compared to single administration and/or vehicle infusion. In addition, administration of MSCs is associated with promotion of interhemispheric connectivity through the CC in the chronic phase of cerebral infarction.

    DOI: 10.3171/2021.8.JNS21687

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  • Postoperative spinal cord ischaemia: magnetic resonance imaging and clinical features International journal

    Naomi Yasuda, Yosuke Kuroda, Toshiro Ito, Masanori Sasaki, Shinichi Oka, Ryo Ukai, Keitaro Nakanishi, Takuma Mikami, Tsuyoshi Shibata, Ryo Harada, Shuichi Naraoka, Takeshi Kamada, Nobuyoshi Kawaharada

    European Journal of Cardio-Thoracic Surgery   60 ( 1 )   164 - 174   2021.7

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    OBJECTIVES: Ischaemic spinal cord injury (SCI) is one of the most serious complications of aortic surgery. Ischaemic SCIs occur due to various aetiologies, and prediction of the risk is difficult. Magnetic resonance imaging (MRI) is useful to detect the details of spinal cord infarction. There are few studies about MRI for evaluating ischaemic SCI after cardiovascular surgery and aortic events. We report 9 cases of postoperative ischaemic SCI and analyse their MRI features. METHODS: T2-weighted MRI scans of 9 patients who developed ischaemic SCI due to cardiovascular surgery and aortic events between 2012 and 2017 were evaluated. RESULTS: In all patients, high-intensity areas were observed on T2-weighted magnetic resonance images. The site of infarction was the thoracic spinal cord level (9 cases) and additionally at the lumbar spinal cord level (5 cases). The area of infarction area was categorized based on the arterial territory: anterior spinal artery territory (3 cases), posterior spinal artery territory (2 cases), spinal sulcal artery territory (1 case) and artery of Adamkiewicz territory (3 cases). CONCLUSIONS: MRI revealed the infarction sites in all cases and the differences in the infarction patterns in each case. MRI could thus be useful for investigating the aetiology of ischaemic SCI following aortic surgeries and events.

    DOI: 10.1093/ejcts/ezaa476

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  • Repeated infusion of mesenchymal stem cells maintain the condition to inhibit deteriorated motor function, leading to an extended lifespan in the SOD1G93A rat model of amyotrophic lateral sclerosis. International journal

    Hirotoshi Magota, Masanori Sasaki, Yuko Kataoka-Sasaki, Shinichi Oka, Ryo Ukai, Ryo Kiyose, Rie Onodera, Jeffery D Kocsis, Osamu Honmou

    Molecular brain   14 ( 1 )   76 - 76   2021.5

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    Amyotrophic lateral sclerosis (ALS) is a neurodegenerative fatal disorder in which motor neurons within the brain and spinal cord degenerate. A single infusion of mesenchymal stem cells (MSCs) delays disease progression by protecting motor neurons and restoring the blood-spinal cord barrier in the SOD1G93A transgenic ALS rat model. However, the therapeutic effect of a single infusion of MSCs is transient and does not block disease progression. In this study, we demonstrated that repeated administration of MSCs (weekly, four times) increased the survival period, protected motor functions, and reduced deterioration of locomotor activity compared to a single infusion and vehicle infusion, after which rats displayed progressive deterioration of hind limb function. We also compared the days until gait ability was lost in rats and found that the repeated-infused group maintained gait ability compared to the single-infusion and vehicle-infusion groups. These results suggest that repeated administration of MSCs may prevent the deterioration of motor function and extend the lifespan in ALS.

    DOI: 10.1186/s13041-021-00787-6

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  • Intravenous Infusion of Mesenchymal Stem Cells Enhances Therapeutic Efficacy of Reperfusion Therapy in Cerebral Ischemia. International journal

    Ryo Kiyose, Masanori Sasaki, Yuko Kataoka-Sasaki, Masahito Nakazaki, Hiroshi Nagahama, Hirotoshi Magota, Shinichi Oka, Ryo Ukai, Mitsuhiro Takemura, Takahiro Yokoyama, Jeffery D Kocsis, Osamu Honmou

    World neurosurgery   149   e160-e169   2021.5

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    OBJECTIVE: Reperfusion therapy is a standard therapeutic strategy for acute stroke. Non-favorable outcomes are thought to partially result from impaired microcirculatory flow in ischemic tissue. Intravenous infusion of mesenchymal stem cells (MSCs) reduces stroke volume and improves behavioral function in stroke. One suggested therapeutic mechanism is the restoration of the microvasculature. The goal of this study was to determine whether infused MSCs enhance the therapeutic efficacy of reperfusion therapy following stroke in rats. METHODS: First, to establish a transient middle cerebral artery occlusion (MCAO) model displaying approximately identical neurologic function and lesion volume as seen in permanent MCAO (pMCAO) at day 7 after stroke induction, we transiently occluded the MCA for 90, 110, and 120 minutes. We found that the 110-minute occlusion met these criteria and was used as the transient MCAO (tMCAO) model. Next, 4 MCAO groups were used to compare the therapeutic efficacy of infused MSCs: (1) pMCAO+vehicle, (2) tMCAO+vehicle, (3) pMCAO+MSC, and (4) tMCAO+MSC. Our ischemic model was a unique ischemic model system in which both pMCAO and tMCAO provided similar outcomes during the study period in the groups without MSC infusion groups. Behavioral performance, ischemic volume, and regional cerebral blood flow (rCBF) using arterial spin labeling-magnetic resonance imaging and histologic evaluation of microvasculature was performed. RESULTS: The behavioral function, rCBF, and restoration of microvasculature were greater in group 4 than in group 3. Thus, infused MSCs facilitated the therapeutic efficacy of MCA reperfusion in this rat model system. CONCLUSIONS: Intravenous infusion of MSCs may enhance therapeutic efficacy of reperfusion therapy.

    DOI: 10.1016/j.wneu.2021.02.056

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  • Intravenous infusion of mesenchymal stem cells delays disease progression in the SOD1G93A transgenic amyotrophic lateral sclerosis rat model. International journal

    Hirotoshi Magota, Masanori Sasaki, Yuko Kataoka-Sasaki, Shinichi Oka, Ryo Ukai, Ryo Kiyose, Rie Onodera, Jeffery D Kocsis, Osamu Honmou

    Brain research   1757   147296 - 147296   2021.4

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    ALS is a devastating neurodegenerative disease with few curative strategies. Both sporadic and familial ALS display common clinical features that show progressive paralysis. The pathogenesis remains unclear, but disruption of the blood-spinal cord barrier (BSCB) may contribute to the degeneration of motor neurons. Thus, restoration of the disrupted BSCB and neuroprotection for degenerating motor neurons could be therapeutic targets. We tested the hypothesis that an intravenous infusion of MSCs would delay disease progression through the preservation of BSCB function and increased expression of a neurotrophic factor, neurturin, in SOD1G93A ALS rats. When the open-field locomotor function was under 16 on the Basso, Beattie, and Bresnahan (BBB) scoring scale, the rats were randomized into two groups; one received an intravenous infusion of MSCs, while the other received vehicle alone. Locomotor function was recorded using BBB scoring and rotarod testing. Histological analyses, quantitative reverse transcription-polymerase chain reaction (qRT-PCR), were performed. The MSC group exhibited reduced deterioration of locomotor activity compared to the vehicle group, which displayed progressive deterioration of hind limb function. We observed the protection of motor neuron loss and preservation of microvasculature using Evans blue leakage and immunohistochemical analyses in the MSC group. Confocal microscopy revealed infused green fluorescent protein+ (GFP+) MSCs in the spinal cord, and the GFP gene was detected by nested PCR. Neurturin expression levels were significantly higher in the MSC group. Thus, restoration of the BSCB and the protection of motor neurons might be contributing mechanisms to delay disease progression in SOD1G93A ALS rats.

    DOI: 10.1016/j.brainres.2021.147296

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  • Recurrence Interval Within 1 Year Leads to Death in Patients with Grade 2 Meningioma International journal

    Ryo Ukai, Masahiko Wanibuchi, Katsuya Komatsu, Yusuke Kimura, Yukinori Akiyama, Takeshi Mikami, Nobuhiro Mikuni

    World Neurosurgery   142   e58 - e65   2020.10

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    OBJECTIVE: Grade 2 meningioma is more likely to recur than grade 1 meningioma. Recurrence decreases overall survival in patients with grade 2 meningioma. However, the clinical course of grade 2 meningioma with several repeated recurrences is poorly understood. The purpose of this study was to clarify the clinical characteristics of grade 2 meningioma with repeated recurrences. METHODS: This study included 28 patients with grade 2 meningioma treated at our institution from January 1994 to December 2017. The relationship between survival and factors including age, sex, number of recurrences, malignant transformation, radiation therapy, tumor location, MIB-1 labeling index, Simpson grade, Karnofsky Performance Status, and surgical interval were analyzed. RESULTS: The average age at the initial operation was 53.4 years. The number of recurrences was 3.7 times on average during the follow-up of 113.9 months after the initial operation. An increasing number of recurrences resulted in shortening of the surgical interval, increase in the MIB-1 labeling index, and decrease in Karnofsky Performance Status. In fatal cases, the average surgical interval before death was approximately 1 year. Three factors were related to poor prognosis: number of recurrences (odds ratio, 1.620; P = 0.030), malignant transformation (odds ratio, 10.625; P = 0.019), and high MIB-1 labeling index (odds ratio, 1.089; P = 0.044). CONCLUSIONS: Shortening of the surgical interval within 1 year because of multiple recurrences led to death in patients with grade 2 meningioma. Malignant transformation was the most potent among the poor prognostic factors.

    DOI: 10.1016/j.wneu.2020.05.145

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  • Arterial transit artifacts observed by arterial spin labeling in Moyamoya disease International journal

    Ryo Ukai, Takeshi Mikami, Hiroshi Nagahama, Masahiko Wanibuchi, Yukinori Akiyama, Kei Miyata, Nobuhiro Mikuni

    Journal of Stroke and Cerebrovascular Diseases   29 ( 9 )   105058 - 105058   2020.9

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    OBJECTIVES: Arterial spin labeling (ASL) is a magnetic resonance imaging (MRI) technique used to assess cerebral perfusion. When tissue perfusion is impaired, such as in Moyamoya disease, a hyperintense band called the arterial transit artifact (ATA) may occur, which interferes with accurate measurements on ASL-MRI. In this study, we evaluated the correlation of ATAs with magnetic resonance angiography (MRA) and single-photon emission computed tomography (SPECT) imaging results in Moyamoya disease. The aim of our study was to elucidate the pathophysiology of ATAs and risk factors for high ATA scores. MATERIALS AND METHODS: This retrospective study included 28 patients (56 hemispheres) with Moyamoya disease treated at our institution. MRI, MRA, ASL perfusion, and N-isopropyl-[123I] b-iodoamphetamine (123I-IMP) SPECT were performed. In order to semi-quantitatively evaluate the degree of ATA, the ATA scores were measured according to the number of hyperintense signal bands in the cerebral cortex. The relationship between the ATA scores and clinical and radiological factors were analyzed. RESULTS: Regional cerebral blood flow (rCBF) determined with ASL weakly correlated with that determined by 123I-IMP SPECT (ρ=0.31, p=0.027). There was no significant association between the ATA scores and rCBF values determined with 123I-IMP SPECT (p=0.872, 0.745, 0.743 at PLD1000 (post-labeling delay), 1500, and 2000, respectively). However, there was a significant correlation between ATA scores and MRA scores (ρ=0.427 p=0.001; ρ=0.612 p=0.001; ρ=0.563 p=0.001 at PLD1000, 1500, and 2000, respectively). An analysis of patient background characteristics revealed a significantly higher incidence of high ATA scores in female patients, patients with high MRA scores, and patients with a distinguishable ivy sign. A multivariate analysis confirmed that female sex, high MRA score, and presence of an ivy sign were risk factors for high ATA scores. CONCLUSION: ATA scores were moderately correlated with MRA scores, and presence of an ivy sign was the most predictive factor for high ATA scores. A high ATA score determined using ASL in a patient with Moyamoya disease might suggest an advanced disease stage and a reduction in cerebrovascular reserve capacity.

    DOI: 10.1016/j.jstrokecerebrovasdis.2020.105058

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  • Natural Y-shaped radial artery graft bypass for a complex middle cerebral artery aneurysm: A case report. International journal

    Ryota Sato, Takeshi Mikami, Hime Suzuki, Akinori Yamamura, Yusuke Kimura, Ryo Ukai, Tomoaki Tamada, Yuka Kawata, Yukinori Akiyama, Nobuhiro Mikuni

    Journal of stroke and cerebrovascular diseases : the official journal of National Stroke Association   29 ( 7 )   104853 - 104853   2020.7

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    Giant thrombosed middle cerebral artery (MCA) aneurysms are difficult to treat and sometimes require complex revascularization using allografts. We describe a technical method using revascularization with a natural Y-shaped graft that provides a normal variation for a complex MCA aneurysm. A 65-year-old man with a giant thrombosed MCA aneurysm presented with right hemiparesis and aphasia. The patient had a history of clipping surgery for the ipsilateral side of the MCA aneurysm 25 years before, and a de novo aneurysm developed over the previous 18 years. For the giant thrombosed aneurysm, trapping and revascularization were performed. A natural radial artery Y-graft was used as the graft and anastomosed to both M2 trunks. The symptoms improved after surgery, and the patient was discharged 3 weeks later. This is the first report of a double-barrel bypass using a natural Y-graft. This method attained a normal variation, and the flow of the Y-graft was physiological. For the radical cure of giant thrombosed MCA aneurysms, multiple revascularizations might be required. With this natural Y-graft, complex transpositions could be avoided.

    DOI: 10.1016/j.jstrokecerebrovasdis.2020.104853

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  • Predictive factors for acute thrombogenesis occurring immediately after bypass procedure for moyamoya disease. International journal

    Takeshi Mikami, Hime Suzuki, Ryo Ukai, Katsuya Komatsu, Yukinori Akiyama, Masahiko Wanibuchi, Kiyohiro Houkin, Nobuhiro Mikuni

    Neurosurgical review   43 ( 2 )   609 - 617   2020.4

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    Extracranial-to-intracranial (EC-IC) bypass surgery is an effective treatment for patients with moyamoya disease and other conditions. Some patients with moyamoya disease have a risk of acute thrombogenesis at the anastomotic site just after bypass surgery. The purpose of this study was to study risk factors of acute thrombogenesis and determine effective countermeasures. This study included 48 patients (66 EC-IC bypass procedures) with moyamoya disease and 52 controls (54 procedures) without moyamoya disease. The development of acute thrombogenesis was compared between the moyamoya disease and control groups. In the moyamoya disease group, clinical and radiological characteristics were assessed with respect to acute thrombogenesis. In the patients with acute thrombogenesis, causes of technical problems were retrospectively examined. The incidence of acute thrombogenesis was significantly higher in the moyamoya disease group than those in the control group. In the moyamoya disease group, acute thrombogenesis was observed in seven patients. In the moyamoya disease group, the magnetic resonance angiography (MRA) scores were significantly higher in patients with acute thrombogenesis than those in the patients without acute thrombogenesis. In the multivariate analysis, the predictive factor of acute thrombogenesis in moyamoya disease was a high MRA score (odds ratio, 2.336; p = 0.009). During EC-IC bypass surgery for moyamoya disease, acute thrombogenesis should be considered to obtain a high patency rate, particularly in patients with high MRA scores. Acute thrombogenesis will not influence morbidity if proper countermeasures are followed; therefore, the prediction and recognition of white thrombus are important for a successful bypass surgery.

    DOI: 10.1007/s10143-019-01086-4

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  • Moyamoya disease with epileptic nystagmus: A case report. International journal

    Chie Nakayama, Takeshi Mikami, Ryo Ukai, Ryohei Chiba, Rei Enatsu, Hime Suzuki, Toru Hirano, Nobuhiro Mikuni

    Journal of clinical neuroscience : official journal of the Neurosurgical Society of Australasia   70   251 - 254   2019.12

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    Epileptic nystagmus is a quick, repetitive, jerky movement of the eyeball caused by seizure activity, which is unaccompanied by other ictal phenomena. We report a case of moyamoya disease with epileptic nystagmus. A 23-year-old woman presented with a headache and transient hemiparesis on her left side. Magnetic resonance imaging showed no ischemic or hemorrhagic stroke lesions. Digital subtraction angiography confirmed stenosis of the terminal portion of the right internal carotid artery and the formation of moyamoya vessels on the right side. 123I-N-isopropyl-iodoamphetamine (123I-IMP) single photon emission computed tomography (SPECT) showed decreased uptake in the right basal ganglia, frontal, and parietal regions. After electroencephalography (EEG) and a hyperventilation test were performed, nystagmus appeared and was accompanied with a declining level of consciousness. Ictal EEG during an attack showed no epileptiform discharge. Moreover, the patient sometimes experienced simultaneous upper limb-shaking and gelastic attacks. After superficial temporal artery to middle cerebral artery bypass surgery was performed on the right side, symptom frequency and duration gradually decreased. Decreased 123I-IMP SPECT blood flow in the right frontal region is considered a mechanism that causes the onset of epileptic nystagmus. It is presumed that the attack was caused by an ischemic abnormality in the saccade region of the frontal eye field. Moreover, revascularization can effectively treat the symptoms of moyamoya disease.

    DOI: 10.1016/j.jocn.2019.08.069

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  • Inflammation promotes progression of thrombi in intracranial thrombotic aneurysms. Reviewed International journal

    Hime Suzuki, Takeshi Mikami, Tomoaki Tamada, Ryo Ukai, Yukinori Akiyama, Akinori Yamamura, Kiyohiro Houkin, Nobuhiro Mikuni

    Neurosurgical review   43 ( 6 )   1565 - 1573   2019.11

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    DOI: 10.1007/s10143-019-01184-3

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  • Pseudoaneurysm presenting around polytetrafluoroethylene fiber following microvascular decompression: A case report and literature review. International journal

    Tomoaki Tamada, Takeshi Mikami, Syoichi Komura, Hime Suzuki, Ryo Ukai, Shintaro Sugita, Tadashi Hasegawa, Nobuhiro Mikuni

    Journal of clinical neuroscience : official journal of the Neurosurgical Society of Australasia   63   231 - 234   2019.5

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    We report the first case of pseudoaneurysm associated with polytetrafluoroethylene fiber used in microvascular decompression (MVD). A 62-year-old female who had undergone MVD for hemifacial spasm 30 years ago presented with a 4-month history of progressive facial palsy. Computed tomography angiography revealed a large thrombosed aneurysm originating from the right posterior inferior cerebellar artery and having a mass effect upon the pons. The aneurysm was treated by trapping and bypass procedure. Intraoperatively, the pseudoaneurysm adhered to the dura mater, and the thrombus contained a large amount of polytetrafluoroethylene fiber. The cause and management of pseudoaneurysm after MVD is discussed.

    DOI: 10.1016/j.jocn.2019.01.041

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  • Flattening the curvature of synthetic materials to relieve scalp skin tension in cranioplasty. International journal

    Takeshi Mikami, Hime Suzuki, Ryo Ukai, Yusuke Kimura, Kei Miyata, Yukinori Akiyama, Masahiko Wanibuchi, Nobuhiro Mikuni

    Journal of clinical neuroscience : official journal of the Neurosurgical Society of Australasia   61   196 - 200   2019.3

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    BACKGROUND: Scalp tissue shrinkage and volume contraction is a major problem in cranioplasty, and sometimes a tissue expander must be set before cranioplasty. The procedure for placing scalp expanders is cumbersome. In this study, we present a method for flattening the curvature of synthetic materials to relieve scalp skin tension and discuss the feasibility and limitations of the method. METHODS: A total of 25 cranioplasty patients were included in this study. The optimal degree of curvature flattening for each piece of bone substitute material was determined based on cosmetic considerations and the extent of encephalomalacia or atrophy due to primary disease. In this series, the correlation between the degree of curvature flattening and the size or location of the bone flap was considered, and the amount of scalp surface area that could be obtained through curvature flattening was estimated. RESULTS: The median degree of curvature flattening was 5.0 mm. The degree of curvature flattening showed moderate correlation with the rate of change in the area of synthetic material achieved through curvature flattening (p < 0.001). The 21 cases of fronto-temporal craniectomy were divided into two groups according to the distance from the midline. There was a statistically significant difference between these two groups in degree of flattening curvature. CONCLUSIONS: In the present cranioplasty series using synthetic materials, curvature flattening was a non-invasive and convenient method for skin closure. This method can be beneficial especially in patients requiring a larger craniotomy including convexity regions.

    DOI: 10.1016/j.jocn.2018.10.032

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  • Surgical Anatomy of Rats for the Training of Microvascular Anastomosis. International journal

    Takeshi Mikami, Hime Suzuki, Ryo Ukai, Katsuya Komatsu, Yusuke Kimura, Yukinori Akiyama, Masahiko Wanibuchi, Nobuhiro Mikuni

    World neurosurgery   120   e1310-e1318   2018.12

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    BACKGROUND: Microvascular anastomosis is an essential procedure in neurosurgery, but the opportunity to perform the surgery has gradually decreased for neurosurgeons. Therefore, training is necessary for obtaining and maintaining the skills required for the procedure. We describe the detailed anatomy of cervical and femoral regions in rats and discuss the advantages for practicing microvascular anastomosis. METHODS: Cervical regions of Sprague-Dawley rats were dissected under intraperitoneal anesthesia. The step-by-step anatomic description was documented using a high-resolution charge-coupled device image sensor and recording systems. Using this model, temporal occlusion time and patency were measured, and these measures were compared between the trainee and trainer groups. The number of times the training needs to be completed to attain competency in the bypass procedure was estimated. RESULTS: After exposing the carotid triangle, a half-ring was created by end-to-side anastomosis. Anastomosis was performed at the common carotid artery using the contralateral side of the carotid artery as a graft. The cutoff value for the temporal occlusion time was 79.3 minutes in the receiver operating characteristic curve based on a target temporal occlusion time for beginners determined during the training. CONCLUSIONS: Using a living animal model, a trainee has the opportunity to learn not only anastomotic techniques but also hemostatic control as well as overcoming mental strain during surgery. Living animal models are important in training because the fidelity of a living animal model is superior to nonliving models. Applying training using a half-ring model contributes to safe and efficient surgery.

    DOI: 10.1016/j.wneu.2018.09.071

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  • Influence of hemodynamics on enlarged perivascular spaces in atherosclerotic large vessel disease. International journal

    Takeshi Mikami, Tomoaki Tamada, Hime Suzuki, Ryo Ukai, Masahiko Wanibuchi, Nobuhiro Mikuni

    Neurological research   40 ( 12 )   1021 - 1027   2018.12

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    OBJECTIVES: Enlarged perivascular spaces (EPVS) are often observed in small vessel disease on T2-weighted images. However, their role in ischemic conditions caused by cerebral large vessel disease remains unclear. We evaluated EPVS in patients with hemodynamic compromise associated with atherosclerotic large vessel disease and aimed to identify the pathophysiology of EPVS. METHODS: We examined 28 adults with atherosclerotic large vessel disease. EPVS numbers in the basal ganglia and the centrum semiovale were assessed. For each affected hemisphere, the total numbers of EPVS were compared with those on the unaffected side. EPVS in the impaired hemodynamics group were compared with those in the unimpaired hemodynamics group. Moreover, EPVS were compared in the presence/absence of large stroke. RESULTS: The number of EPVS was significantly increased on the affected side in the centrum semiovale (p = 0.023), particularly in the impaired hemodynamics group (p = 0.006). Moreover, in the small stroke subgroup of the impaired hemodynamics group, the number of EPVS was significantly increased on the affected side (p = 0.002), although this number was insignificant in the large ischemic stroke subgroup. CONCLUSIONS: The number of EPVS was increased in patients with atherosclerotic large vessel disease with hemodynamic compromise and decreased in the presence of a large stroke. EPVS might act as fluid absorbers in a hemodynamically compromised state until the occurrence of an ischemic stroke.

    DOI: 10.1080/01616412.2018.1509827

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  • 浮遊血栓による軽症内頸動脈閉塞の2例

    山岡 歩, 飯星 智史, 宮田 圭, 千葉 遼平, 木村 友亮, 鵜飼 亮, 三上 毅, 鰐渕 昌彦, 三國 信啓

    脳血管内治療   3 ( Suppl. )   S293 - S293   2018.11

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    Ichushi

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  • Interdisciplinary Prevention and Management of Wound-Related Complications in Extracranial-to-Intracranial Bypass Surgery. International journal

    Rintaro Yokoyama, Takeshi Mikami, Ryo Ukai, Katsuya Komatsu, Yusuke Kimura, Hime Suzuki, Toshimi Honma, Toru Hirano, Tamotsu Saito, Ken Yamashita, Takatoshi Yotsuyanagi, Kiyohiro Houkin, Nobuhiro Mikuni

    World neurosurgery   115   247 - 253   2018.7

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    Extracranial-to-intracranial (EC-IC) bypass surgery may be necessary in patients with moyamoya disease and other ischemic conditions. However, there is a potential risk of wound-related complications in some cases. In this study, we report our approach to the prevention of wound-related complications in EC-IC bypass. Technical considerations and pitfalls of surgery are also discussed. This study included 89 patients with ischemic-onset moyamoya disease and atherosclerotic disease who underwent 108 superficial temporal artery (STA)-to-middle cerebral artery bypass procedures. Our study emphasized 3 major features. First, 3-dimensional simulation imaging was used to confirm STA anatomy. Second, the STA was meticulously dissected on the epigaleal layer to protect the galeal layer. Third, scalp skin ischemia was taken into consideration at each step until skin closure. There was no neurologic morbidity or mortality in this series. There were 2 cases of major wound-related complications requiring plastic surgical intervention, and 4 cases of minor complications that were treated conservatively. In major complication cases, the scalp defect was treated with pedicle flap reconstruction. In EC-IC bypass surgery, interdisciplinary management involving neurosurgery, plastic surgery, and radiology should reduce wound-related complications and achieve safe surgery.

    DOI: 10.1016/j.wneu.2018.04.167

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  • 前交通動脈瘤に対するbasal interhemispheric approach 到達距離についての検討 Reviewed

    鰐渕 昌彦, 金 相年, 鵜飼 亮, 山内 朋裕, 杉野 寿哉, 今井 哲秋, 茂木 洋晃, 大瀧 雅文

    CI研究   31 ( 2 )   105 - 108   2009.9

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    Ichushi

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  • Mesenchymal Stem Cells Derived from Peripheral Blood Protects against Ischemia

    Ryo Ukai

    Journal of Neurotrauma   24   508 - 520   2007.3

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    DOI: 10.1089/neu.2006.0161

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MISC

  • Neurogenesisと神経再生医療 脳梗塞に対する骨髄幹細胞移植による脳循環の改善

    本望 修, 鵜飼 亮, 原田 邦明, 宝金 清博

    脳循環代謝   18 ( 4 )   189 - 191   2006.12

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Research Projects

  • 脳梗塞に対するsmall Extracellular Vesicles静脈投与療法の治療メカニズムの解明

    Grant number:25K12323  2025.4 - 2028.3

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    鵜飼 亮, 佐々木 祐典, 横山 貴裕, 中崎 公仁, 本望 修

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    Grant amount:\4550000 ( Direct Cost: \3500000 、 Indirect Cost:\1050000 )

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  • 脊髄損傷に対する骨髄間葉系幹細胞移植による、脳脊髄の可塑性亢進メカニズムの解析

    Grant number:24K12269  2024.4 - 2027.3

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    岡 真一, 鵜飼 亮, 横山 貴裕, 中崎 公仁

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    Grant amount:\4550000 ( Direct Cost: \3500000 、 Indirect Cost:\1050000 )

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  • 慢性期脳梗塞に対する骨髄幹細胞治療における至適リハビリ条件の探索

    Grant number:24K14250  2024.4 - 2027.3

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    山下 達郎, 佐々木 雄一, 佐々木 祐典, 鵜飼 亮, 岡 真一, 佐々木 優子, 本望 修, 中崎 公仁

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    Grant amount:\4550000 ( Direct Cost: \3500000 、 Indirect Cost:\1050000 )

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  • Therapeutic strategies for cerebral infarction and spinal cord injury by activating plasticity throughout the central nervous system

    Grant number:23K24447  2024.4 - 2025.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

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    Grant amount:\5460000 ( Direct Cost: \4200000 、 Indirect Cost:\1260000 )

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  • 脊髄損傷に対する骨髄幹細胞による治療メカニズムに占めるミトコンドリア機能の解析

    Grant number:23K08547  2023.4 - 2026.3

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    佐々木 祐典, 鵜飼 亮, 岡 真一, 佐々木 優子, 本望 修

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    Grant amount:\4810000 ( Direct Cost: \3700000 、 Indirect Cost:\1110000 )

    脊髄損傷後に神経症状の増悪をもたらす二次損傷の病態解明は重要である。近年、二次損傷に占めるミトコンドリアの役割に注目が集まっており、脊髄損傷後に生じるミトコンドリアの機能不全によるATP合成不全が惹起するエネルギー代謝不全、活性酸素の産生増加、神経毒性の増加などが神経症状の増悪に関与すると報告されている。一方、我々はこれまで、脊髄損傷ラットモデルおよび脊髄損傷患者に対する骨髄幹細胞の経静脈的投与 (MSC治療)による運動機能の改善を報告してきた。
    この中で、MSC治療後に、ミトコンドリアの代謝機能に関連する遺伝子群が存在することを見出した。本研究では、脊髄損傷に対しMSC治療を実施し、①損傷局所および②大脳皮質の神経系細胞内におけるミトコンドリアの機能の変化を詳細に解析することを計画した。これらの解析を通じて、MSC治療により惹起される神経機能の回復に貢献するメカニズムにミトコンドリアが強く関与していることを明らかにし、次世代の治療法の開発に展開することを最終的な目標として、これまでに雄性SDラット (250-300g)に実験的脊髄損傷 (IH-0400 Impactor)を作製後にMSCの経静脈的投与を行い (MSC群)、運動機能を評価した。さらに、細胞外フラックスアナライザー (Xfe96, Agilent)を用いて、ミトコンドリアエネルギー代謝機能である酸素消費速度 (oxygen consumption rate: OCR)および細胞外酸性化速度(extracellular acidification rate: ECAR)を測定の予備的実験を開始した。

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  • Development of innovative treatments using next-generation exosomes for cerebral infarction

    Grant number:22K09239  2022.4 - 2025.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

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    Grant amount:\4160000 ( Direct Cost: \3200000 、 Indirect Cost:\960000 )

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  • 脳梗塞と脊髄損傷に対する中枢神経系全域の可塑性賦活化による治療戦略の検討

    Grant number:22H03188  2022.4 - 2025.3

    日本学術振興会  科学研究費助成事業  基盤研究(B)

    本望 修, 佐々木 祐典, 鵜飼 亮, 岡 真一, 佐々木 優子

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    Grant amount:\16900000 ( Direct Cost: \13000000 、 Indirect Cost:\3900000 )

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  • 健康寿命延長に関与する骨髄幹細胞の自己治癒能と全身の恒常性維持

    Grant number:22K11807  2022.4 - 2025.3

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    佐々木 優子, 佐々木 祐典, 鵜飼 亮, 岡 真一, 本望 修

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    Grant amount:\4160000 ( Direct Cost: \3200000 、 Indirect Cost:\960000 )

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  • Reconstructed neural circuits following intravenous infusion of mesenchymal stem cells in a rat model of experimental cerebral ischemia

    Grant number:21K09104  2021.4 - 2024.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

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    Grant amount:\3900000 ( Direct Cost: \3000000 、 Indirect Cost:\900000 )

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  • 骨髄幹細胞治療における機能活性モニタリングと機能予後バイオマーカーの開発

    Grant number:21K09131  2021.4 - 2024.3

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    中村 秀之, 佐々木 祐典, 佐々木 優子, 本望 修, 鵜飼 亮, 横山 貴裕

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    Grant amount:\4160000 ( Direct Cost: \3200000 、 Indirect Cost:\960000 )

    我々は、脳梗塞に対する骨髄間葉系幹細胞(Mesenchymal stemcells:MSC)の経静脈的投与(MSC治療)による良好な治療効果を報告してきた。我々のこれまでの基礎研究において、ラット脳梗塞モデルを用いた実験を行い、MSC治療後の血漿中BDNFは、投与されたMSCの機能的バイオマーカーとなりうる可能性があることを報告した (Nakamura et al., 2017) 本研究では、血漿中BDNFの推移が、機能活性の指標に留まらず、脳梗塞の機能予後と相関するのかどうかを検証することを計画しており、さらに、機能活性や機能予後を示す新しいバイオマーカーを探索することを目的としている。これまでに、①医師主導治験患者の血漿中BDNF測定、および、②ラットMCAOモデルにMSCを投与し、血漿中BDNFと機能予後の相関を検討することで、MSC治療における機能活性モニタリングによる機能予後バイオマーカーの開発を行っている。①では、札幌医科大学附属病院に入院した患者から、経時的に採血を行い、血漿を分離し、ELISA法にてBDNFを測定し、ELIZA法によるBDNFの測定は、BDNF E-max immunoassay system (Promega) を用いる。②では、ラット脳梗塞モデルとして、中大脳動脈永久閉塞(MCAO)モデルを用いる。MCAOモデルは、Intraluminal thread methodを用いて作製し、脳梗塞発症6時間後にラットMSCを経静脈的に投与し、末梢血を採取し、保存血から血漿を分離し、①同様にELISA法にてBDNFを測定する。観察期間中、並行して、トレッドミルによってラットの最大走行速度を測定し、運動機能および動物用MRIを用いて脳梗塞の体積を経時的に評価する。観察期間終了時に、血漿中BDNFと運動機能から機能予後解析を継続して行うこととしている。以上のように、補助金は適切に使用し、予定通り進行中である。

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  • 脊髄損傷に対する骨髄間葉系幹細胞移植による、脳脊髄での可塑性亢進メカニズムの解析

    Grant number:21K09182  2021.4 - 2024.3

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    岡 真一, 佐々木 祐典, 横山 貴裕, 本望 修, 鵜飼 亮

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    Grant amount:\4160000 ( Direct Cost: \3200000 、 Indirect Cost:\960000 )

    令和3年度、当院にMSC治療を目的に入院した脊髄損傷症例は、おおよそ20例を数え、順調に症例数を積み重ねている状況である。全ての脊髄損傷症例の脳、脊髄のMRIデータを検証した。撮像時期は、原則、入院時、投与直前、 投与1ヶ月後、投与3ヶ月後、投与6ヶ月後の時点としていたかが、投与条件に当てはまらず投与を受けなかった症例や入院病床の調整の関係で、MRIの撮像を出来ない症例も散見された。特に脳DTIデータを用いたコネクトーム解析では、標準化の方法、使用する脳atlasの選択、ROI間の接続強度の閾値設定など、症例間でばらつきのない均一な解析を実施するためのパラメータの設定の検討を引き続き実施している。さらに、T1WI、T2WIなどを用いて大脳皮質厚や体積などの構造学的なデータ解析も同時に実施し、コネクトーム解析で得られた結果との比較も実施している。また、脊髄MRIでは、個々の症例においてDTI解析の可否や、定量的な解析を実施し、データの蓄積を行っている。撮像データの解析、特にDTIデータの解析を目的として高機能のパーソナルコンピューターを別途購入し、専用のアプリケーションにて解析を実施した。
    ラット脊髄損傷モデルに対して骨髄間葉系幹細胞(MSC)の経静脈投与することによって、中枢神経系にどのような神経回路の再構築(plasticity)が生じるかを検証するため、ラット胸髄Th9レベルの脊髄損傷モデルを作成し、動物用MRIでの撮像条件の最適化を検討した。8-9週齢のSDラットを購入し、ラット用脊髄損傷作 成装置(IH-0400 Impactor; PSI社製)を用いて、安定した損傷部位を作成できることを確認した。その後、動物実験用高磁場MRI装置にて、脳、脊髄おける各種プロトコール(T1、T2、DTI等)での撮像を行い、撮像条件の調整を行い最適な撮像条件の検討を行った。

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  • 慢性期脳梗塞に対する骨髄幹細胞とリハビリ併用による脳のplasticityの解析

    Grant number:21K11194  2021.4 - 2024.3

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    山下 達郎, 佐々木 祐典, 本望 修, 鵜飼 亮

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    Grant amount:\4160000 ( Direct Cost: \3200000 、 Indirect Cost:\960000 )

    近年、骨髄間葉系幹細胞(MSC)を用いた細胞療法(MSC治療)は、再生医療の分野で脳梗塞や脊髄損傷に対する新しい治療方法として注目されている。MSC移植の治療メカニズムは、①サイトカインによる神経栄養作用、②血液脳関門機能の修復、③血管新生、④脱髄軸索の再有髄化、⑤神経再生による脳のplasticityの調節、と多段階・協奏的に作用することが判明しており、臨床上の神経症状の回復も、ほぼこれと平行して進むことが判明している。我々は、既にラット急性期脳梗塞モデルに対してMSC移植にリハビリを併用すると、リハビリ単独群やMSC移植単独群よりも、さらに高い運動機能の回復が得られるこという報告をした。MSCは、1)梗塞巣周囲に集積し、2)脳梗塞体積を減少させ、3)脳梗塞周辺領域のシナプス新生を誘導し、4)脳梁萎縮を抑制、するが、リハビリを併用すると、脳の可塑性のさらなる亢進を誘導することが可能となり、より高い治療効果が得られることを明らかにした。さらに慢性期脳梗塞モデルに対するMSC移植は、運動機能が改善されることを明らかにした。急性期と慢性期で病態は異なるが、MSCが発揮する多彩な治療メカニズムを考えると、慢性期にMSC移植とリハビリを併用することでさらなる治療効果が期待できると考えられる。本研究は、慢性期脳梗塞モデルに対するMSC移植にリハビリを併用することで、機能改善をもたらすメカニズムを脳の可塑性の変化に注目して解析することを目的としている。現在までに、本研究費によって、実験的慢性期脳梗塞に対する治療効果の行動学的解析、MRI解析、神経トレーサーを用いた神経解剖学的解析などを行っている。以上のように、補助金は補助条件に従って、有効に使用されている。

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  • 脳梗塞に対する骨髄幹細胞治療における分子メカニズムに基づく至適リハビリ条件の探索

    Grant number:20K11212  2020.4 - 2023.3

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    佐々木 雄一, 佐々木 祐典, 岡 真一, 佐々木 優子, 本望 修, 山下 達郎, 鵜飼 亮

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    Grant amount:\4290000 ( Direct Cost: \3300000 、 Indirect Cost:\990000 )

    近年、骨髄間葉系幹細胞(MSC)を用いた細胞療法(MSC治療)は、再生医療の分野で脳梗塞や脊髄損傷に対する新しい治療方法として注目されている。我々は、既にラット脳梗塞モデルに対するMSC移植にリハビリを併用すると、リハビリ単独群やMSC移植単独群よりも、さらに高い運動機能の回復が得られるこという報告をし た。MSCは、1)梗塞巣周囲に集積し、2)脳梗塞体積を減少させ、3)脳梗塞周辺領域のシナプス新生を誘導し、4)脳梁萎縮を抑制、するが、リハビリを併用すると、脳の可塑性のさらなる亢進を誘導することが可能となり、より高い治療効果が得られることを明らかにした。しかし、実臨床に目を向けると、MSC治療とリハビリの至適なプロトコルの確立はなされておらず、様々な議論が絶えない状況である。一方、最近のリハビリの動向として、早期リハビリや高負荷・高頻度のリハビリの有用性、さらに、日常生活における適切な環境因子に注目が集まっている。本研究は、MSC移植に併用する適切な運動(リハビリ)の種類や頻度・強度 とともに、飼育環境も考慮し、回復経過を分子メカニズムなど様々な視点から詳細に比較解析することで、MSC移植の治療効果を最大限に引き出すための新しいリハビリ方法を確立することを目的としている。特に、運動強度と運動時間をなどの負荷量に注目し、脳の可塑性のパターンの変化、神経回路の再構築、シナプス新生などの観点から解析を進めている。現在までに、本研究費によって、各運動負荷における実験的脳梗塞に対する治療効果の行動学的解析、MRI解析、神経トレーサーを用いた神経解剖学的解析などを行っている。以上のように、補助金は補助条件に従って、有効に使用されている。

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  • Extended lifespan induced by mesenchymal stem cells

    Grant number:19K11794  2019.4 - 2022.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    Kataoka-Sasaki Yuko

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    Grant amount:\4420000 ( Direct Cost: \3400000 、 Indirect Cost:\1020000 )

    In this study, we demonstrated that intravenous infusion of MSCs increased the survival rate in rat models of spontaneously hypertensive (stroke-prone) in which organs including kidney, brain, heart, and liver are damaged during aging due to spontaneous hypertension. Intravenous infusion of MSC may extend lifespan in addition to preventing the deterioration of motor and cognitive functions in the animal model.

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