MF研究者総覧

研究者データベース

詳細
七戸　秀夫(シチノヘ　ヒデオ)
北海道大学病院　医療・ヘルスサイエンス研究開発機構
教授
researchmap個人ページ
Last Updated :2024/05/09

基本情報

所属

  • 北海道大学病院　医療・ヘルスサイエンス研究開発機構

職名

  • 教授

学位

  • 医学博士（北海道大学）

J-Global ID

研究キーワード

  • Bone marrow stromal cell   Spinal cord injury   Differentiation   Neuroprotection   Cerebral stroke   Migration   脊髄損傷   骨髄間質細胞   脳梗塞   サイトカイン   中枢神経再生医療   PET   バイオマテリアル   光イメージング   幹細胞移植   Regenerative medicine   Diffuse axonal injury   再生医療   Cerebral infarction   Functional restoration   Regeneration   機能回復   transplantation   脳挫傷   脳損傷   biomaterial   神経再生   抗血管新生因子   

研究分野

  • 人文・社会 / 哲学、倫理学 / 生命倫理、研究倫理、臨床研究、脳神経倫理学
  • ライフサイエンス / 脳神経外科学 / 脳卒中、再生医療

担当教育組織

職歴

  • 2021年04月 - 現在 北海道大学大学院文学研究院 応用倫理・応用哲学研究教育センター 研究員
  • 2021年04月 - 現在 北海道大学病院 医療・ヘルスサイエンス研究開発機構 臨床研究監理センター 准教授
  • 2020年07月 - 現在 株式会社RAINBOW 研究開発担当取締役
  • 2017年01月 - 2021年03月 北海道大学病院 臨床研究監理部長 准教授
  • 2016年09月 - 2016年12月 北海道大学病院 臨床研究開発センター 准教授
  • 2015年04月 - 2016年08月 北海道大学大学院医学研究科 脳神経外科 特任助教
  • 2015年02月 - 2016年08月 (独) 医薬品医療機器総合機構 再生医療製品等審査部 特任職員(非常勤)
  • 2010年08月 - 2015年03月 北海道大学病院 脳神経外科 助教

研究活動情報

論文

  • Masahito Kawabori, Satoshi Kuroda, Hideo Shichinohe, Kaoru Kahata, Souichi Shiratori, Satoshi Ikeda, Taisuke Harada, Kenji Hirata, Khin Khin Tha, Masato Aragaki, Shunsuke Terasaka, Yoichi M Ito, Naoki Nishimoto, Shunsuke Ohnishi, Ichiro Yabe, Kohsuke Kudo, Kiyohiro Houkin, Miki Fujimura
    Med (New York, N.Y.) 2024年03月18日 
    BACKGROUND: Ischemic stroke is one of the leading causes of death and neurological disability worldwide, and stem cell therapy is highly expected to reverse the sequelae. This phase 1/2, first-in-human study evaluated the safety, feasibility, and monitoring of an intracerebral-transplanted magnetic resonance imaging (MRI)-trackable autologous bone marrow stromal cell (HUNS001-01) for patients with subacute ischemic stroke. METHODS: The study included adults with severe disability due to ischemic stroke. HUNS001-01 cultured with human platelet lysates and labeled with superparamagnetic iron oxide was stereotactically transplanted into the peri-infarct area 47-64 days after ischemic stroke onset (dose: 2 or 5 × 107 cells). Neurological and radiographic evaluations were performed throughout 1 year after cell transplantation. The trial was registered at UMIN Clinical Trial Registry (number UMIN000026130). FINDINGS: All seven patients who met the inclusion criteria successfully achieved cell expansion, underwent intracerebral transplantation, and completed 1 year of follow-up. No product-related adverse events were observed. The median National Institutes of Health Stroke Scale and modified Rankin scale scores before transplantation were 13 and 4, which showed improvements of 1-8 and 0-2, respectively. Cell tracking proved that the engrafted cells migrated toward the infarction border area 1-6 months after transplantation, and the quantitative susceptibility mapping revealed that cell signals at the migrated area constantly increased throughout the follow-up period up to 34% of that of the initial transplanted site. CONCLUSIONS: Intracerebral transplantation of HUNS001-01 was safe and well tolerated. Cell tracking shed light on the therapeutic mechanisms of intracerebral transplantation. FUNDING: This work was supported by the Japan Agency for Medical Research and Development (AMED; JP17bk0104045 and JP20bk0104011).
  • Soichiro Takamiya, Masahito Kawabori, Tsukasa Kitahashi, Kentaro Nakamura, Yuki Mizuno, Hironobu Yasui, Yuji Kuge, Aki Tanimori, Yasuyuki Takamatsu, Kohei Yuyama, Hideo Shichinohe, Miki Fujimura
    Stem Cells International 2022 1 - 10 2022年07月31日 
    Background. Due to the lack of effective therapies, stem cell transplantation is an anticipated treatment for chronic intracerebral hemorrhage (ICH), and higher cell survival and engraftment are considered to be the key for recovery. Mesenchymal stromal cells (MSCs) compounded with recombinant human collagen type I scaffolds (CellSaics) have a higher potential for cell survival and engraftment compared with solo-MSCs, and we investigated the validity of intracerebral transplantation of CellSaic in a chronic ICH model. Methods. Rat CellSaics (rCellSaics) were produced by rat bone marrow-derived MSC (rBMSCs). The secretion potential of neurotrophic factors and the cell proliferation rate were compared under oxygen-glucose deprivation (OGD) conditions. rCellSaics, rBMSCs, or saline were transplanted into the hollow cavity of a rat chronic ICH model. Functional and histological analyses were evaluated, and single-photon emission computed tomography for benzodiazepine receptors was performed to monitor sequential changes in neuronal integrity. Furthermore, human CellSaics (hCellSaics) were transplanted into a chronic ICH model in immunodeficient rats. Antibodies neutralizing brain-derived neurotrophic factor (BDNF) were used to elucidate its mode of action. Results. rCellSaics demonstrated a higher secretion potential of trophic factors and showed better cell proliferation in the OGD condition. Animals receiving rCellSaics displayed better neurological recovery, higher intracerebral BDNF, and better cell engraftment; they also showed a tendency for less brain atrophy and higher benzodiazepine receptor preservation. hCellSaics also promoted significant functional recovery, which was reversed by BDNF neutralization. Conclusion. Intracerebral transplantation of CellSaics enabled neurological recovery in a chronic ICH model and may be a good option for clinical application.
  • Masahito Kawabori, Hideo Shichinohe, Satoshi Kuroda, Kiyohiro Houkin
    STROKE 52 2021年03月 [査読有り]
  • Masahito Kawabori, Aki Tanimori, Shinri Kitta, Hideo Shichinohe, Kiyohiro Houkin
    Stem cells international 2021 9796010 - 9796010 2021年 [査読有り]
     
    [This corrects the article DOI: 10.1155/2020/4085617.].
  • 脳梗塞に対する新規治療の開発-血栓回収療法時代のアンメットメディカルニーズと医薬品開発戦略- 主幹動脈閉塞脳梗塞患者に対する幹細胞脳内移植治験(RAINBOW研究)
    川堀 真人, 七戸 秀夫, 黒田 敏, 宝金 清博
    脳循環代謝 32 1 52 - 52 (一社)日本脳循環代謝学会 2020年11月
  • 脳梗塞急性期患者への自家BMSC脳内投与による再生治療(Phase 1)とPhase 2に向けた取り組み
    川堀 真人, 七戸 秀夫, 黒田 敏, 宝金 清博
    脳循環代謝 32 1 60 - 60 (一社)日本脳循環代謝学会 2020年11月
  • 脳神経細胞治療・遺伝子治療の実践 脳出血慢性期モデルに対する骨髄幹細胞+足場材製剤(CellSaic)の有効性
    高宮 宗一朗, 川堀 真人, 七戸 秀夫, 中村 健太郎, 北橋 宗, 岩澤 玲子, 古川 友子, 宝金 清博
    脳循環代謝 32 1 68 - 68 (一社)日本脳循環代謝学会 2020年11月
  • Masahito Kawabori, Hideo Shichinohe, Satoshi Kuroda, Kiyohiro Houkin
    International journal of molecular sciences 21 19 2020年10月06日 
    Despite recent developments in innovative treatment strategies, stroke remains one of the leading causes of death and disability worldwide. Stem cell therapy is currently attracting much attention due to its potential for exerting significant therapeutic effects on stroke patients. Various types of cells, including bone marrow mononuclear cells, bone marrow/adipose-derived stem/stromal cells, umbilical cord blood cells, neural stem cells, and olfactory ensheathing cells have enhanced neurological outcomes in animal stroke models. These stem cells have also been tested via clinical trials involving stroke patients. In this article, the authors review potential molecular mechanisms underlying neural recovery associated with stem cell treatment, as well as recent advances in stem cell therapy, with particular reference to clinical trials and future prospects for such therapy in treating stroke.
  • Chengbo Tan, Zifeng Wang, Miao Zheng, Songji Zhao, Hideo Shichinohe, Kiyohiro Houkin
    Journal of Nippon Medical School = Nippon Ika Daigaku zasshi 88 3 228 - 237 2020年08月31日 
    BACKGROUND: Stroke is a leading cause of death and disability worldwide. Recently, brain secondary damage has been hypothesized to be a key aggravating element in an ischemic cascade. However, the interaction between cerebral infarction and immune organs has yet to be fully understood. In this study, we investigated the changes in the rat brain, spleen, thymus, mesenteric lymph node, and liver at 3, 7, and 13 days after transient middle cerebral artery occlusion (tMCAO) by immunohistochemistry. MATERIAL AND METHODS: Rat models of stroke were made by tMCAO. Functional assessment was performed 3 h, and 1, 3, 5, 7, 9, 11, and 13 days after MCAO. Rat organs were harvested for 2,3,5-triphenyltetrazolium chloride staining and Immunohistochemistry. RESULTS: The CD8α+ T cells was found to decrease in the spleen, thymus, mesenteric lymph node, and liver, whereas it increased in the brain. Those of Iba1+ and CD68+ macrophages were decreased in the spleen, thymus, and mesenteric lymph node, whereas they were elevated in the brain and liver. Ki67+ cells showed the same characteristics as macrophages, and increased numbers of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) positive apoptotic cells were found in the spleen, mesenteric lymph node, liver, and brain. CONCLUSIONS: The present results demonstrated that stroke is a systemic disease, which not only affects the brain, but also induces responses of immune organs. On the basis of these results, a systemic treatment might be a good strategy for clinical stroke care.
  • 川堀 真人, 七戸 秀夫, 黒田 敏, 寳金 清博
    臨床画像 36 4月増刊 141 - 150 (株)メジカルビュー社 2020年04月 [査読有り][通常論文]
     
    ＜文献概要＞脳梗塞・頭部外傷・脊髄損傷・Parkinson病など中枢神経疾患に対する細胞治療・再生医療が,基礎研究から治験の段階(bench to bed)に入っている。これらは既存治療ではなしえなかった中枢神経を再生・回復させる治療法として大きく期待されている。本報告では,脳血管障害に対する研究結果および今後の課題について概説する。
  • Masahito Kawabori, Aki Tanimori, Shinri Kitta, Hideo Shichinohe, Kiyohiro Houkin
    Stem cells international 2020 4085617 - 4085617 2020年 [査読有り][通常論文]
     
    Cell therapy for central nervous system (CNS) disorders is beginning to prove its safety and efficiency. Intraparenchymal transplantation can be an option for cell delivery; however, one concern regarding this method is that the transplantation cannula may cause additional brain injuries. These include vessel damage, which results in brain hemorrhage, and clogging of the cannula by brain debris and/or cell clusters, which requires replacement of the cannula or forced injection causing jet flow of the cell suspension. We compared cannulas for cell delivery used in clinical trials, the Pittsburg and Mizuho cannulas, to a newly designed one, MK01, to assess their usability. MK01 has a spherical-shaped tip with a fan-like open orifice on the side of the cannula, which prevents vessel damage, clogging of brain debris, and jet flow phenomenon. We compared the extent of rat cervical and abdominal arterial damage with the cannula, the amount of debris in the cannula, the force needed to cause jet flow, and cell viability. While the viability of cells passed through the cannulas was almost the same among cannulas (approximately 95%), the Pittsburg cannula caused cervical arterial injury and subsequent hemorrhage, as it required a significantly smaller force to penetrate the arterial wall. Moreover, the Pittsburg cannula, but not the Mizuho and MK01 cannulas, showed high frequency of brain debris in the needle tip (approximately 80%) after brain puncture. While jet flow of the injection liquid was observed even when using smaller forces in the Pittsburg and Mizuho cannulas, MK01 constantly showed low jet flow occurrence. Thus, MK01 seems to be safer than the previously reported cannulas, although further investigation is necessary to validate its safety for clinical use.
  • 脳梗塞亜急性期に対する自家骨髄間質細胞の脳内投与 第一相治験:RAINBOW研究
    川堀 真人, 七戸 秀夫, 黒田 敏, 寳金 清博
    脳循環代謝 31 1 83 - 83 (一社)日本脳循環代謝学会 2019年11月 [査読有り][通常論文]
  • 川堀 真人, 七戸 秀夫, 黒田 敏, 宝金 清博
    日本臨床 77 6 945 - 953 (株)日本臨床社 2019年06月
  • 川堀 真人, 七戸 秀夫, 黒田 敏, 寳金 清博
    神経治療学 36 6 S128 - S128 日本神経治療学会 2019年
  • Tan C, Zhao S, Higashikawa K, Wang Z, Kawabori M, Abumiya T, Nakayama N, Kazumata K, Ukon N, Yasui H, Tamaki N, Kuge Y, Shichinohe H, Houkin K
    EJNMMI research 8 1 35 - 35 2018年05月 [査読有り][通常論文]
     
    BACKGROUND: The potential application of bone marrow stromal cell (BMSC) therapy in stroke has been anticipated due to its immunomodulatory effects. Recently, positron emission tomography (PET) with [18F]DPA-714, a translocator protein (TSPO) ligand, has become available for use as a neural inflammatory indicator. We aimed to evaluate the effects of BMSC administration after transient middle cerebral artery occlusion (MCAO) using [18F]DPA-714 PET. The BMSCs or vehicle were administered intravenously to rat MCAO models at 3 h after the insult. Neurological deficits, body weight, infarct volume, and histology were analyzed. [18F]DPA-714 PET was performed 3 and 10 days after MCAO. RESULTS: Rats had severe neurological deficits and body weight loss after MCAO. Cell administration ameliorated these effects as well as the infarct volume. Although weight loss occurred in the spleen and thymus, cell administration suppressed it. In both vehicle and BMSC groups, [18F]DPA-714 PET showed a high standardized uptake value (SUV) around the ischemic area 3 days after MCAO. Although SUV was increased further 10 days after MCAO in both groups, the increase was inhibited in the BMSC group, significantly. Histological analysis showed that an inflammatory reaction occurred in the lymphoid organs and brain after MCAO, which was suppressed in the BMSC group. CONCLUSIONS: The present results suggest that BMSC therapy could be effective in ischemic stroke due to modulation of systemic inflammatory responses. The [18F]DPA-714 PET/CT system can accurately demonstrate brain inflammation and evaluate the BMSC therapeutic effect in an imaging context. It has great potential for clinical application.
  • Houkin K, Shichinohe H, Abe K, Arato T, Dezawa M, Honmou O, Horie N, Katayama Y, Kudo K, Kuroda S, Matsuyama T, Miyai I, Nagata I, Niizuma K, Sakushima K, Sasaki M, Sato N, Sawanobori K, Suda S, Taguchi A, Tominaga T, Yamamoto H, Yamashita T, Yoshimine T, Working Group, for Guidelines on Development of Cell-Based, Products for the Treatment of Cerebral Infarction
    Stroke 49 4 e145 - e152 2018年04月 [査読有り][通常論文]
  • Miyamoto M, Nakamura K, Shichinohe H, Yamauchi T, Ito M, Saito H, Kawabori M, Osanai T, Sasaki T, Houkin K, Kuroda S
    Stem cells international 2018 4829534 - 4829534 2018年 [査読有り][通常論文]
     
    Bone marrow stromal cell (BMSC) transplantation has the therapeutic potential for ischemic stroke. However, it is unclear which delivery routes would yield both safety and maximal therapeutic benefits. We assessed whether a novel recombinant peptide (RCP) sponge, that resembles human collagen, could act as a less invasive and beneficial scaffold in cell therapy for ischemic stroke. BMSCs from green fluorescent protein-transgenic rats were cultured and Sprague-Dawley rats were subjected to permanent middle cerebral artery occlusion (MCAo). A BMSC-RCP sponge construct was transplanted onto the ipsilateral intact neocortex 7 days after MCAo. A BMSC suspension or vehicle was transplanted into the ipsilateral striatum. Rat motor function was serially evaluated and histological analysis was performed 5 weeks after transplantation. The results showed that BMSCs could proliferate well in the RCP sponge and the BMSC-RCP sponge significantly promoted functional recovery, compared with the vehicle group. Histological analysis revealed that the RCP sponge provoked few inflammatory reactions in the host brain. Moreover, some BMSCs migrated to the peri-infarct area and differentiated into neurons in the BMSC-RCP sponge group. These findings suggest that the RCP sponge may be a promising candidate for animal protein-free scaffolds in cell therapy for ischemic stroke in humans.
  • Daisuke Shimbo, Takeo Abumiya, Kota Kurisu, Toshiya Osanai, Hideo Shichinohe, Naoki Nakayama, Ken Kazumata, Hideki Nakamura, Hiroshi Shimuzu, Kiyohiro Houkin
    JOURNAL OF STROKE & CEREBROVASCULAR DISEASES 26 12 2994 - 3003 2017年12月 [査読有り][通常論文]
     
    Background: The development of cerebral infarction after transient ischemia is attributed to postischemic delayed hypoperfusion in the microvascular region. In the present study, we assessed the microvascular perfusion capacity of infused liposome-encapsulated hemoglobin (LEH) in a therapeutic approach for transient middle cerebral artery occlusion (tMCAO). Methods: Two-hour middle cerebral artery occlusion rats were immediately subjected to intra-arterial infusion of LEH (LEH group) or saline (vehicle group) or no treatment (control group), and then to recanalization. Neurological findings, infarct and edema progression, microvascular endothelial dysfunction, and inflammatory reactions were compared between the 3 groups after 24 hours of reperfusion. Microvascular perfusion in the early phase of reperfusion was evaluated by hemoglobin immunohistochemistry and transmission electron microscopy. Results: The LEH group achieved significantly better results in all items evaluated than the other groups. Hemoglobin immunohistochemistry revealed that the number of hemoglobin-positive microvessels was significantly greater in the LEH group than in the other groups (P < .01), with microvascular perfusion being more likely in narrow microvessels (<= 5 mu m in diameter). An electron microscopic examination revealed that microvessels in the control group were compressed and narrowed by swollen astrocyte end-feet, whereas those in the LEH group had a less deformed appearance and contained LEH particles and erythrocytes. Conclusion: The results of the present study demonstrated that the infusion of LEH reduced infarctions after tMCAO with more hemoglobin-positive and less deformed microvessels at the early phase of reperfusion, suggesting that the superiority of the microvascular perfusion of LEH mediates its neuroprotective effects.
  • Hideo Shichinohe, Masahito Kawabori, Hiroaki Iijima, Tuyoshi Teramoto, Takeo Abumiya, Naoki Nakayama, Ken Kazumata, Shunsuke Terasaka, Teruyo Arato, Kiyohiro Houkin
    BMC NEUROLOGY 17 1 179  2017年09月 [査読有り][通常論文]
     
    Background: Stroke is a leading cause of death and disability, and despite intensive research, few treatment options exist. However, a recent breakthrough in cell therapy is expected to reverse the neurological sequelae of stroke. Although some pioneer studies on the use of cell therapy for treating stroke have been reported, certain problems remain unsolved. Recent studies have demonstrated that bone marrow stromal cells (BMSCs) have therapeutic potential against stroke. We investigated the use of autologous BMSC transplantation as a next-generation cell therapy for treating stroke. In this article, we introduce the protocol of a new clinical trial, the Research on Advanced Intervention using Novel Bone marrOW stem cell (RAINBOW). Methods/design: RAINBOW is a phase 1, open-label, uncontrolled, dose-response study, with the primary aim to determine the safety of the autologous BMSC product HUNS001-01 when administered to patients with acute ischemic stroke. Estimated enrollment is 6-10 patients suffering from moderate to severe neurological deficits. Approximately 50 mL of the bone marrow is extracted from the iliac bone of each patient 15 days or later from the onset. BMSCs are cultured with allogeneic human platelet lysate (PL) as a substitute for fetal calf serum and are labeled with superparamagnetic iron oxide for cell tracking using magnetic resonance imaging (MRI). HUNS00101 is stereotactically administered around the area of infarction in the subacute phase. Each patient will be administered a dose of 20 or 50 million cells. Neurological scoring, MRI for cell tracking, F-18-fuorodeoxyglucose positron emission tomography, and I-123-Iomazenil singlephoton emission computed tomography will be performed for 1 year after the administration. Discussion: This is a first-in-human trial for HUNS001-01 to the patients with acute ischemic stroke. We expect that intraparenchymal injection can be a more favorable method for cell delivery to the lesion and improvement of the motor function than intravenous infusion. Moreover, it is expected that the bio-imaging techniques can clarify the therapeutic mechanisms.
  • Ken Kazumata, Khin Khin Tha, Haruto Uchino, Tohru Shiga, Hideo Shichinohe, Masaki Ito, Naoki Nakayama, Takeo Abumiya
    JOURNAL OF NEUROSURGERY 127 2 260 - 269 2017年08月 [査読有り][通常論文]
     
    OBJECTIVE After revascularization surgery, hyperperfusion and ischemia are associated with morbidity and mortality in adult moyamoya disease (MMD). However, structural changes within the brain following revascularization surgery, especially in the early postsurgical period, have not been thoroughly studied. Such knowledge may enable improved monitoring and clinical management of hyperperfusion and ischemia in MMD. Thus, the objective of this study was to investigate the topographic and temporal profiles of cerebral perfusion and related white matter microstructural changes following revascularization surgery in adult MMD. METHODS The authors analyzed 20 consecutive surgeries performed in 17 adults. Diffusion imaging in parallel with serial measurements of regional cerebral blood flow (rCBF) using SPECT was performed. Both voxel-based and region of-interest analyses were performed, comparing neuroimaging parameters of postoperative hemispheres with those of preoperative hemispheres at 4 different time points within 2 weeks after surgery. RESULTS Voxel-based analysis showed a distinct topographic pattern of cerebral perfusion, characterized by increased rCBF in the basal ganglia for the first several days and gradually increased rCBF in the lateral prefrontal cortex over 1 week (p < 0.001). Decreased rCBF was also observed in the lateral prefrontal cortex, occipital lobe, and cerebellum contralateral to the surgical hemisphere (p < 0.001). Reduced fractional anisotropy (FA) and axial diffusivity (AD), as well as increased radial diffusivity (RD), were demonstrated in both the anterior and posterior limbs of the internal capsule (p < 0.001). Diffusion parameters demonstrated the greatest changes in both FA and RD on Days 1-2 and in AD on Days 3-6; FA, RD, and AD recovered to preoperative levels on Day 14. Patients with transient neurological deteriorations (TNDs), as compared with those without, demonstrated greater increases in rCBF in both the lateral prefrontal cortex and striatum as well as smaller FAs in the posterior limb of the internal capsule (p < 0.05). CONCLUSIONS The excessively increased rCBF and the recovery process were heterogeneous across brain regions, demonstrating a distinct topographic pattern during the initial 2 weeks following revascularization surgery in MMD. Temporary impairments in the deep white matter tract and immediate postoperative ischemia were also identified. The study results characterized postoperative brain perfusion as well as the impact of revascularization surgery on the brain microstructure. Notably, rCBF and white matter changes correlated to TNDs, suggesting that these changes represent potential neuroimaging markers for tracking tissue structural changes associated with hyperperfusion during the acute postoperative period following revascularization surgery for MMD.
  • Masaaki Hokari, Naoki Nakayama, Ken Kazumata, Toshiya Osanai, Hideo Shichinohe, Takeo Abumiya, Kiyohiro Houkin
    NEUROLOGIA MEDICO-CHIRURGICA 57 3 122 - 127 2017年03月 [査読有り][通常論文]
     
    There are no reports on the outcomes of clippings in patients who receive immunosuppressants, for example, due to connective tissue diseases or following organ transplantation. We thoroughly reviewed these cases focusing on the perioperative management phase. The study included 11 patients with intracranial aneurysms who were taking immunosuppressants; between 2007 and 2014. We performed 12 clipping surgeries. Their clinical records were reviewed for age and gender, aneurysms' location and size, perioperative management of the immunosuppressive drugs, and surgical complications. The study included nine females and two males, aged between 52 and 71 years (mean 60.1 +/- 8.5 years). The clinical presentation in five cases was subarachnoid hemorrhage (SAH); the aneurysm was incidentally diagnosed in six patients (7 aneurysms). The reasons for taking immunosuppressants were autoimmune disorder in nine patients and liver transplantation in two patients. Daily intake of oral immunosuppressants for the patients with liver transplantation was discontinued for 2-4 days, and no infectious complications were evidenced. The weekly course of immunosuppressive drugs for the patients with autoimmune disorder was continued in eight of nine patients. Caution must be exercised when considering the suitability of clipping for patients taking immunosuppressants, but surgery outcomes are generally favorable; when operative treatment is required, we believe it to be comparatively safe, if the perioperative management is conducted in close collaboration with the relevant departments.
  • Masaki Ito, Hideo Shichinohe, Kiyohiro Houkin, Satoshi Kuroda
    JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE 11 2 375 - 381 2017年02月 [査読有り][通常論文]
     
    Bone marrow stromal cells (BMSC) transplantation enhances functional recovery after cerebral infarct, but the optimal delivery route is undetermined. This study was aimed to assess whether a novel cell-sheet technology non-invasively serves therapeutic benefits to ischemic stroke. First, the monolayered cell sheet was engineered by culturing rat BMSCs on a temperature-responsive dish. The cell sheet was analysed histologically and then transplanted onto the ipsilateral neocortex of rats subjected to permanent middle cerebral artery occlusion at 7 days after the insult. Their behaviours and histology were compared with those in the animals treated with direct injection of BMSCs or vehicle over 4 weeks post-transplantation. The cell sheet was 27.9+/-8.0 mu m thick and was composed of 9.8+/-2.4x10(5) cells. Cell sheet transplantation significantly improved motor function when compared with the vehicle-injected animals. Histological analysis revealed that the BMSCs were densely distributed to the neocortex adjacent to the cerebral infarct and expressed neuronal phenotype in the cell sheet-transplanted animals. These findings were almost equal to those for the animals treated with direct BMSC injection. The attachment of the BMSC sheet to the brain surface did not induce reactive astrocytes in the adjacent neocortex, although direct injection of BMSCs profoundly induced reactive astrocytes around the injection site. These findings suggest that the BMSCs in cell sheets preserve their biological capacity of migration and neural differentiation. Cell-sheet technology may enhance functional recovery after ischaemic stroke, using a less invasive method. Copyright (C) 2014 John Wiley & Sons, Ltd.
  • Hisayasu Saito, Michiyuki Miyamoto, Hideo Shichinohe, Kiyohiro Houkin, Satoshi Kuroda
    Cell Therapy against Cerebral Stroke: Comprehensive Reviews for Translational Researches and Clinical Trials 111 - 119 2017年01月01日 [査読有り][通常論文]
     
    Cell transplantation therapy has been expected to promote functional recovery in various kinds of central nervous system (CNS) disorders, including cerebral stroke. However, there are several concerns to be resolved before clinical application of cell therapy for CNS disorders. The issues include the development of imaging techniques to monitor the response of the host CNS. It would be essential to establish functional bio-imaging technique serially and noninvasively validating the effects of cell therapy on the host CNS in order to achieve clinical application of cell therapy for cerebral stroke. Nuclear imaging technique is one of the most useful methods to assess the functional change in various kinds of CNS disorders, including cerebral stroke. Very recently, using a small-animal SPECT/CT apparatus, we could serially visualize the effects of BMSC transplantation on the distribution of 123I-IMZ in the infarct brain of the living rodents longitudinally and noninvasively. Furthermore, we serially assessed local glucose metabolism in the rats subjected to permanent MCA occlusion and found that BMSC transplantation significantly enhances the recovery in the peri-infarct area, using smallanimal 18F-FDG PET/CT system. The BMSCs may enhance the recovery of local glucose metabolism by improving neuronal integrity in the peri-infarct area, when directly transplanted into the infarct brain. Although there are few studies that indicate the utility of imaging techniques to monitor the response of the host CNS after cell therapy and further investigation is needed, 123I-IMZ SPECT and 18FFDG PET may be promising modalities to assess the therapeutic benefits of cell therapy for ischemic stroke without subjective bias in clinical situation.
  • Satoshi Kuroda, Hideo Shichinohe, Kiyohiro Houkin
    Cell Therapy against Cerebral Stroke: Comprehensive Reviews for Translational Researches and Clinical Trials 15 - 25 2017年01月01日 [査読有り][通常論文]
     
    In this article, the authors review recent advancements and perspective on cell therapy for ischemic stroke with bone marrow-derived cells, including bone marrow stromal cells (BMSCs) and multilineage-differentiating stress-enduring (Muse) cells. They can be easily isolated from the patients themselves and transplanted into them without any ethical and immunological problem. Animal experiments have shown that direct transplantation of these adult stem cells significantly enhances the recovery of motor function in various types of neurological disorders, including ischemic stroke. They aggressively migrate toward the damaged tissue and proliferate in the host brain. The BMSCs may contain heterogeneous subpopulations and contribute to functional recovery through multiple mechanisms, including neuroprotection, inflammatory modulation, cell fusion, and neural differentiation. On the other hands, Muse cells may promote functional recovery after ischemic stroke by reorganizing the infarct brain.
  • Kurisu K, Osanai T, Kazumata K, Nakayama N, Abumiya T, Shichinohe H, Shimoda Y, Houkin K
    Neurologia medico-chirurgica 56 12 745 - 752 2016年12月 [査読有り][通常論文]
     
    Although ultrasound (US) guidance for venous access is becoming the "standard of care" for preventing access site complications, its feasibility for arterial access has not been fully investigated, especially in the neuro-interventional population. We conducted the first prospective cohort study on US-guided femoral artery access during neuro-interventional procedure. This study included 64 consecutive patients who underwent US-guided femoral artery access through 66 arterial access sites for diagnostic and/or neuro-interventional purposes. The number of attempts required for both the sheath insertion and the success of anterior wall puncture were recorded. In addition, the occurrence of major complications and hematoma formation on the arterial access site examined by US were statistically analyzed. The median number of attempts was 1 (1-2) and first-pass success rate was 63.6%. Anterior wall puncture was achieved in 98.5%. In one case (1.5%), a pseudoaneurysm was observed. In all cases, US clearly depicted a common femoral artery (CFA) and its bifurcation. Post-procedural hematoma was detected in 13 cases (19.7%), most of which were "tiny" or "moderate" in size. Low body mass index and antiplatelet therapy were the independent risk factors for access site hematoma. The US-guided CFA access was feasible even in neuro-interventional procedure. The method was particularly helpful in the patients with un-palpable pulsation of femoral arteries. To prevent arterial access site hematoma, special care should be taken in patients with low body mass index and who are on antiplatelet therapy.
  • Furukawa K, Abumiya T, Sakai K, Hirano M, Osanai T, Shichinohe H, Nakayama N, Kazumata K, Hida K, Houkin K
    Journal of stroke and cerebrovascular diseases : the official journal of National Stroke Association 25 11 2762 - 2769 Elsevier 2016年11月 [査読有り][通常論文]
     
    Background and Purpose: High blood viscosity causes blood stagnation and subsequent pathological thrombotic events, resulting in the development of ischemic stroke. We hypothesize that the contribution of blood viscosity may differ among ischemic stroke subtypes based on specific pathological conditions. We tried to verify this hypothesis by measuring blood viscosity in acute ischemic stroke patients using a newly developed electromagnetic spinning sphere (EMS) viscometer. Methods: Measurements in acute ischemic stroke patients were performed 4 times during admission and data were compared with those obtained from 100 healthy outpatient volunteers. Results: We enrolled 92 patients (cardioembolism [CE]: 25, large-artery atherosclerosis [LAA]: 42, and small artery occlusion [SAO]: 25) in this study. Comparisons of blood viscosity between the ischemic stroke subgroups and control group revealed that blood viscosity at the date of admission was significantly higher in the SAO group (5.37±1.11 mPa・s) than in the control group (4.66±0.72 mPa・s) (p<0.01). Among all subtype groups showing a reduction in blood viscosity after 2 weeks, the SAO group showed the highest and most significant reduction, indicating that SAO patients had the most concentrated blood at the onset. Conclusions: Blood viscosity was significantly increased in the SAO group at the date of admission, which indicated the contribution of dehydration to the onset of ischemic stroke. The importance of dehydration needs to be emphasized more in the pathogenesis of SAO. The clinical application of the EMS viscometer is promising for understanding and differentiating the pathogenesis of ischemic stroke.
  • Kurisu K, Abumiya T, Ito M, Gekka M, Osanai T, Shichinohe H, Nakayama N, Kazumata K, Houkin K
    Brain research 1651 95 - 103 2016年11月 [査読有り][通常論文]
     
    The robust neuroprotective effects of transarterial regional hypothermia have been demonstrated in the typical transient middle cerebral artery occlusion (tMCAO) model, but have not yet been tested in other ischemic stroke models, even though clinical ischemic conditions are diverse. In order to clarify these effects in a different ischemic stroke model, we employed a rat model of permanent MCAO (pMCAO) with transient collateral hypoperfusion (tCHP), which was achieved by direct MCA ligation through craniotomy and 1-h bilateral common carotid artery occlusion at the beginning of pMCAO. The infusion of 20ml/kg of 4°C cold saline (CS) or 37°C warm saline (WS) into the ipsilateral internal carotid artery (ICA) was performed for 15min in intra- or post-tCHP. Neurological scores, infarct/edema volumes, and neuronal apoptosis and reactive gliosis were compared between the CS and WS groups and a non-infusion control group after 48h of reperfusion. Although brain temperatures were only reduced by 2-3°C for 15min, the CS group had significantly better neurological scores, smaller infarct/edema volumes, and less penumbral neuronal apoptosis and reactive gliosis than the control and WS groups. The post-tCHP CS group exhibited prominent neuroprotective effects, even though infarct volumes and neuronal apoptosis were reduced less than those in the intra-tCHP CS group. In conclusion, we demonstrated the neuroprotective effects of transarterial regional hypothermia in an ischemic model of pMCAO with tCHP. Even though MCAO is persistent, cold infusion via the ICA is neuroprotective for the penumbra, suggesting the wider therapeutic application of this therapy.
  • Shichinohe H, Houkin K
    Neurologia medico-chirurgica 56 10 592 - 596 2016年10月 [査読有り][通常論文]
     
    Stroke is still a leading cause of death and disability, and despite intensive research, few treatment options exist. A recent breakthrough in cell therapy is expected to reverse the neurological sequelae of stroke. Although some pioneer studies on the use of cell therapy for the treatment of stroke have been reported, certain problems still remain unsolved. We investigated the use of autologous bone marrow stromal cell (BMSC) transplantation for the treatment of stroke, to develop it as the next-generation cell therapy. In this study, we introduce the preparation of a new clinical trial, the Research on Advanced Intervention using Novel Bone marrow stem cell (RAINBOW) study. The trial will start in 2016, and we hope that it will not only be helpful for treating patients but also for clarifying the therapeutic mechanisms. Moreover, we review stem cell therapeutics as an emerging paradigm in stroke (STEPS) and the guidelines for the development of cell therapy for stroke in the United States as well as introduce the development of new guidelines in Japan. These guidelines are expected to encourage the development of cell therapy for stroke management.
  • Furukawa K, Abumiya T, Sakai K, Hirano M, Osanai T, Shichinohe H, Nakayama N, Kazumata K, Aida T, Houkin K
    Journal of medical engineering & technology 40 6 285 - 292 2016年08月 [査読有り][通常論文]
     
    We herein applied an electromagnetic spinning sphere (EMS) viscometer to the measurement of human blood viscosity for the first time. We collected blood samples from 100 healthy outpatient volunteers in order to analyse viscosity dependence on blood cell parameters and on the shear rate with a simple approximation formula [ηi (γ)\, = Ai γ(- pi) + η0]. Viscosity dependence on blood cell parameters was relatively high at a high shear rate, but became lower as the shear rate decreased. The approximation formula with appropriate parameters of Ai and pi nearly faithfully reproduced actual blood rheological behaviour with a standard deviation of 1.5%. The distributions of Ai and pi values were broad, suggesting that the pattern of viscosity dependence on the shear rate varied with individual differences. The results obtained using the EMS viscometer suggest that blood viscosity values are individual-specific and actual individual measurements are important for understanding rheological conditions.
  • Kurisu K, Abumiya T, Nakamura H, Shimbo D, Shichinohe H, Nakayama N, Kazumata K, Shimizu H, Houkin K
    Neurosurgery 79 1 125 - 134 2016年07月 [査読有り][通常論文]
     
    BACKGROUND: Although transarterial regional hypothermia is an attractive alternative to general hypothermia, its efficacy and underlying mechanisms remain unclear. OBJECTIVE: To confirm transarterial regional hypothermia therapeutic effects on ischemia/reperfusion (I/R) injury and to elucidate the mechanisms responsible. METHODS: The therapeutic effects of transarterial regional hypothermia were initially investigated in 2-hour middle cerebral artery occlusion rats regionally infused with 10°C saline (cold saline group) or 37°C saline (warm saline group) and untreated rats (control group) just before the onset of 24 hours of reperfusion. The time course of infarct and edema progression, inflammatory reactions, microvascular morphological changes, and aquaporin-4 (AQP4) expression was analyzed after 0, 2, 6, and 24 hours of reperfusion. RESULTS: Cold saline infusion only lowered brain temperatures for 30 minutes but mediated strong neuroprotective effects with infarct volume reductions of less than one-third. The time-course analysis revealed the following sequence of ischemia/reperfusion injury-related events in the control group: upregulated expression of AQP4 (2 hours); microvascular narrowing resulting from swollen astrocytic end-feet (2-6 hours); infarct and edema progression, blood-brain barrier disruption, and upregulated expression of intracellular adhesion molecule-1 (6-24 hours); and the activation of other inflammatory reactions (24 hours). These sequential events were inhibited in the cold saline group. CONCLUSION: Transarterial regional hypothermia initially inhibited the acute AQP4 surge and then attenuated microvascular narrowing, blood-brain barrier disruption, and activation of other inflammatory reactions, leading to strong neuroprotective effects. More direct and intensive cooling of the endothelium and its surroundings may contribute to these effects. ABBREVIATIONS: AQP4, aquaporin-4BBB, blood-brain barrierIba1, ionized calcium-binding adapter molecule 1ICA, internal carotid arteryICAM-1, intracellular adhesion molecule-1I/R, ischemia/reperfusionMCAO, middle cerebral artery occlusionMMP-9, matrix metalloproteinase-9.
  • Kazumata K, Tha K.K, Narita H, Shichinohe H, Ito M, Uchino H, Abumiya T
    World Neurosurgery 89 654 - 665.e2 2016年05月 [査読有り][通常論文]
     
    Chronic ischemia in adult moyamoya disease (MMD) reduces the integrity of normal-appearing white matter (WM). We investigated whether covert WM impairment alters large-scale brain networks and specific neural circuits associated with neurocognitive dysfunction in MMD. Forty-six participants (control, n = 23; MMD, n = 23) were examined using diffusion tensor imaging and streamline tractography. Structural connectivity among 90 cortical and subcortical brain regions was evaluated using the mean fractional anisotropy along the fiber tracts. Graph theoretical analysis was used to measure network parameters and inter-regional connectivity. Global network parameters were reduced in patients with MMD, including cluster coefficient (controls vs. MMD: 3.62 ± 0.24 vs. 3.26 ± 0.36; P < 0.0001), characteristic path length (controls vs. MMD: 1.20 ± 0.02 vs. 1.17 ± 0.01; P < 0.001), and small-world property (controls vs. MMD: 3.07 ± 0.18 vs. 2.83 ± 0.27; P < 0.001). Reduced pairwise connectivity was found in prefrontal neural circuits within the middle/inferior frontal gyrus; supplementary motor area; and insular, inferior temporal, and dorsal cingulate cortices. Covert WM microstructural changes in patients with MMD alter large-scale brain networks, as well as lateral prefrontal neural circuits. Evaluation of structural connectivity may be useful to assess the severity of chronic ischemic injury from a network perspective.
  • Shimoda Y, Osanai T, Nakayama N, Ushikoshi S, Hokari M, Shichinohe H, Abumiya T, Kazumata K, Houkin K
    Journal of neurosurgery. Pediatrics 17 3 330 - 335 2016年03月 [査読有り][通常論文]
     
    Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant systemic disorder characterized by the enlargement of capillaries, recurrent nosebleeds, and multiple arteriovenous malformations (AVMs). Although cerebral AVMs are traditionally considered to be congenital lesions, some reports have described de novo AVMs, which suggests that the authors believed them to be dynamic conditions. In this article, the authors describe the case of a 5-year-old boy with HHT in whom a de novo cerebral AVM was detected after a negative MRI result at 5 months. To the authors' knowledge, this is the first report of a de novo AVM in a patient with HHT. In patients with a family history of HHT, de novo AVMs are possible, even when no lesions are detected at the first screening. Therefore, regular screenings need to be performed, and the family should be informed that AVMs could still develop despite normal MRI results.
  • Shimoda Yusuke, Moriwaki Takuya, Nakayama Naoki, Abumiya Takeo, Kazumata Ken, Shichinohe Hideo, Houkin Kiyohiro
    STROKE 47 2016年02月 [査読有り][通常論文]
  • Abumiya Takeo, Furukawa Koji, Sakai Keiji, Hirano Miki, Osanai Toshiya, Shichinohe Hideo, Nakayama Naoki, Kazumata Ken, Aida Toshimitsu, Houkin Kiyohiro
    STROKE 47 2016年02月 [査読有り][通常論文]
  • Shichinohe Hideo, Tan Chengbo, Saito Hisayasu, Miyamoto Michiyuki, Hamauchi Shuji, Abumiya Takeo, Nakayama Naoki, Kazumata Ken, Houkin Kiyohiro, Kuroda Satoshi
    STROKE 47 2016年02月 [査読有り][通常論文]
  • Kurisu Kota, Abumiya Takeo, Ito Masaki, Shichinohe Hideo, Nakayama Naoki, Kazumata Ken, Osanai Toshiya, Houkin Kiyohiro
    STROKE 47 2016年02月 [査読有り][通常論文]
  • Chengbo Tan, Hideo Shichinohe, Zifeng Wang, Shuji Hamauchi, Takeo Abumiya, Naoki Nakayama, Ken Kazumata, Tsuneo Ito, Kohsuke Kudo, Shigeru Takamoto, Kiyohiro Houkin
    Stem Cells International 2016 2016年 [査読有り][通常論文]
     
    Currently, there is increasing interest in human bone marrow stromal cells (hBMSCs) as regeneration therapy against cerebral stroke. The aim of the present study was to evaluate the feasibility and validity of hBMSC cultures with allogeneic platelet lysates (PLs). Platelet concentrates (PC) were harvested from healthy volunteers and made into single donor-derived PL (sPL). The PL mixtures (mPL) were made from three different sPL. Some growth factors and platelet cell surface antigens were detected by enzyme-linked immunosorbent assay (ELISA). The hBMSCs cultured with 10% PL were analyzed for their proliferative potential, surface markers, and karyotypes. The cells were incubated with superparamagnetic iron oxide (SPIO) agents and injected into a pig brain. MRI and histological analysis were performed. Consequently, nine lots of sPL and three mPL were prepared. ELISA analysis showed that PL contained adequate growth factors and a particle of platelet surface antigens. Cell proliferation capacity of PLs was equivalent to or higher than that of fetal calf serum (FCS). No contradiction in cell surface markers and no chromosomal aberrations were found. The MRI detected the distribution of SPIO-labeled hBMSCs in the pig brain. In summary, the hBMSCs cultured with allogeneic PL are suitable for cell therapy against stroke.
  • Takamiya S, Osanai T, Ushikoshi S, Kurisu K, Shimoda Y, Ito Y, Ishi Y, Hokari M, Nakayama N, Kazumata K, Abumiya T, Shichinohe H, Houkin K
    No shinkei geka. Neurological surgery 44 1 39 - 45 2016年01月 [査読有り][通常論文]
     
    Systemic vascular diseases such as fibromuscular dysplasia, Ehlers-Danlos syndrome, Marfan syndrome, and Behçet's disease are known to cause spontaneous dissecting aneurysms of the cervical internal carotid artery. These diseases are generally associated with vascular fragility; therefore, invasive treatments are avoided in many cases of dissecting aneurysms, and a conservative approach is used for the primary disease. Surgical or intravascular treatment may be chosen when aneurysms are progressive or are associated with a high risk of hemorrhage; however, there is no consensus on which treatment is better. We report a case of a dissecting aneurysm of the cervical internal carotid artery in a patient with suspected Behçet's disease, which was treated using stent-assisted coil embolization. A man in his 40's, with suspected Behçet's disease, presented with an enlarged dissecting aneurysm of the right cervical internal carotid artery. The lesion was present for approximately 10 years. We performed stent-assisted coil embolization for the lesion. Post-surgery, no aneurysms were detected with carotid artery echography. Our case report suggests that stent-assisted coil embolization is a promising treatment for dissecting aneurysms of the cervical internal carotid artery. In addition, the procedure demonstrates the utility of carotid artery echograms for examining recanalization after stent-assisted coil embolization.
  • Hamauchi S, Shichinohe H, Uchino H, Yamaguchi S, Nakayama N, Kazumata K, Osanai T, Abumiya T, Houkin K, Era T
    PloS one 11 9 e0163561  2016年 [査読有り][通常論文]
     
    Backgroundand purpose Moyamoya disease (MMD) is a slow, progressive steno-occlusive disease, arising in the terminal portions of the cerebral internal carotid artery. However, the functions and characteristics of the endothelial cells (ECs) in MMD are unknown. We analyzed these features using induced pluripotent stem cell (iPSC)-derived ECs.MethodsiPSC lines were established from the peripheral blood of three patients with MMD carrying the variant RNF213 R4810K, and three healthy persons used as controls. After the endothelial differentiation of iPSCs, CD31(+)CD144(+) cells were purified as ECs using a cell sorter. We analyzed their proliferation, angiogenesis, and responses to some angiogenic factors, namely VEGF, bFGF, TGF-beta, and BMP4. The ECs were also analyzed using DNA microarray and proteomics to perform comprehensive gene and protein expression analysis.ResultsAngiogenesis was significantly impaired in MMD regardless of the presence of any angiogenic factor. On the contrary, endothelial proliferation was not significant between controland MMD-derived cells. Regarding DNA microarray, pathway analysis illustrated that extracellular matrix (ECM) receptor-related genes, including integrin beta 3, were significantly down-regulated in MMD. Proteomic analysis revealed that cytoskeleton-related proteins were downregulated and splicing regulation-related proteins were upregulated in MMD.ConclusionsDownregulation of ECM receptor-related genes may be associated with impaired angiogenic activity in ECs derived from iPSCs from patients with MMD. Upregulation of splicing regulation-related proteins implied differences in splicing patterns between control and MMD ECs.
  • Tan C, Shichinohe H, Wang Z, Hamauchi S, Abumiya T, Nakayama N, Kazumata K, Ito T, Kudo K, Takamoto S, Houkin K
    Stem cells international 2016 6104780 - 6104780 2016年 [査読有り][通常論文]
     
    Currently, there is increasing interest in human bone marrow stromal cells (hBMSCs) as regeneration therapy against cerebral stroke. The aim of the present study was to evaluate the feasibility and validity of hBMSC cultures with allogeneic platelet lysates (PLs). Platelet concentrates (PC) were harvested from healthy volunteers and made into single donor-derived PL (sPL). The PL mixtures (mPL) were made from three different sPL. Some growth factors and platelet cell surface antigens were detected by enzyme-linked immunosorbent assay (ELISA). The hBMSCs cultured with 10% PL were analyzed for their proliferative potential, surface markers, and karyotypes. The cells were incubated with superparamagnetic iron oxide (SPIO) agents and injected into a pig brain. MRI and histological analysis were performed. Consequently, nine lots of sPL and three mPL were prepared. ELISA analysis showed that PL contained adequate growth factors and a particle of platelet surface antigens. Cell proliferation capacity of PLs was equivalent to or higher than that of fetal calf serum (FCS). No contradiction in cell surface markers and no chromosomal aberrations were found. The MRI detected the distribution of SPIO-labeled hBMSCs in the pig brain. In summary, the hBMSCs cultured with allogeneic PL are suitable for cell therapy against stroke.
  • Kazumata K, Tha K.K, Narita H, Ito Y.M, Shichinohe H, Ito M, Uchino H, Abumiya T
    American Journal of Neuroradiology 37 8 1432 - 1439 2016年 [査読有り][通常論文]
     
    BACKGROUND AND PURPOSE: Detecting microstructural changes due to chronic ischemia potentially enables early identification of patients at risk of cognitive impairment. In this study, diffusional kurtosis imaging and diffusion tensor imaging were used to investigate whether the former provides additional information regarding microstructural changes in the gray and white matter of adult patients with Moyamoya disease. MATERIALS AND METHODS: MR imaging (diffusional kurtosis imaging and DTI) was performed in 23 adult patients with Moyamoya disease and 23 age-matched controls. Three parameters were extracted from diffusional kurtosis imaging (mean kurtosis, axial kurtosis, and radial kurtosis), and 4, from DTI (fractional anisotropy, radial diffusivity, mean diffusivity, and axial diffusivity). Voxelwise analysis for these parameters was performed in the normal-appearing brain parenchyma. The association of these parameters with neuropsychological performance was also evaluated. RESULTS: Voxelwise analysis revealed the greatest differences in fractional anisotropy, followed, in order, by radial diffusivity, mean diffusivity, and mean kurtosis. In patients, diffusional kurtosis imaging parameters were decreased in the dorsal deep white matter such as the corona radiata and superior longitudinal fasciculus (P < .01), including areas without DTI abnormality. Superior longitudinal fasciculus fiber-crossing areas showed weak correlations between diffusional kurtosis imaging and DTI parameters compared with tissues with a single-fiber direction (eg, the corpus callosum). Diffusional kurtosis imaging parameters were associated with general intelligence and frontal lobe performance. CONCLUSIONS: Although DTI revealed extensive white matter changes, diffusional kurtosis imaging additionally demonstrated microstructural changes in ischemia-prone deep white matter with abundant fiber crossings. Thus, diffusional kurtosis imaging may be a useful adjunct for detecting subtle chronic ischemic injuries.
  • Shimoda Yusuke, Nakayama Naoki, Hokari Masaaki, Abumiya Takeo, Shichinohe Hideo, Kazumata Ken, Houkin Kiyohiro
    JOURNAL OF NEUROSURGERY 123 2 A526  2015年08月 [査読有り][通常論文]
  • Hokari M, Nakayama N, Kazumata K, Osanai T, Nakamura T, Yasuda H, Ushikoshi S, Shichinohe H, Abumiya T, Kuroda S, Houkin K
    Journal of stroke and cerebrovascular diseases : the official journal of National Stroke Association 24 8 1768 - 1774 2015年08月 [査読有り][通常論文]
     
    BACKGROUND: Carotid endarterectomy (CEA) and carotid stenting (CAS) are beneficial procedures for patients with high-grade cervical carotid stenosis. However, it is sometimes difficult to manage patients with bilateral carotid stenosis. To decide the treatment strategy, one of the most important questions is whether contralateral stenosis increases the risk of patients undergoing CEA. METHODS: This retrospective study included 201 patients with carotid stenosis who underwent a total of 219 consecutive procedures (CEA 189/CAS 30). We retrospectively analyzed outcomes in patients with carotid stenosis who were treated with either CEA or CAS and evaluated whether or not contralateral lesions increases the risk of patients undergoing CEA or CAS. Furthermore, we retrospectively verified our treatment strategy for bilateral carotid stenosis. RESULTS: The incidences of perioperative complications were 5.3% in the CEA patients and 6.7% in the CAS patients, respectively. There was no significant difference between these 2 groups. The existences of contralateral occlusion and/or contralateral stenosis were not associated with perioperative complications in both the groups. There were 32 patients with bilateral severe carotid stenosis (>50%). Of those, 13 patients underwent bilateral revascularizations; CEA followed by CEA in 8, CEA followed by CAS in 3, CAS followed by CEA + coronary artery bpass grafting in 1, and CAS followed by CAS in 1. CONCLUSIONS: Our date showed that the existence of contralateral carotid lesion was not associated with perioperative complications, and most of our cases with bilateral carotid stenosis initially underwent CEA.
  • Shichinohe H, Tan C, Abumiya T, Nakayama N, Kazumata K, Hokari M, Houkin K, Kuroda S
    Brain research 1602 53 - 61 2015年03月 [査読有り][通常論文]
     
    The phosphodiesterase (PDE) 3 inhibitor cilostazol, used as an anti-platelet drug, reportedly can also ameliorate ischemic brain injury. Here, we investigated the effects of cilostazol in a permanent focal ischemia mice model. Male Balb/c mice were subjected to permanent middle cerebral artery occlusion. Mice were then treated with either cilostazol (10 or 20mg/kg) or vehicle administered at 30min and 24h post-ischemia, and infarct volumes were assessed at 48h post-ischemia. Mice treated with 20mg/kg of cilostazol or vehicle were sacrificed at 6h or 24h post-ischemia and immunohistochemistry was used for brain sections. Treatment with 20mg/kg of cilostazol significantly reduced infarct volumes to 70.1% of those with vehicle treatment. Immunohistochemistry results for 8-hydroxydeoxyguanosine (OHdG) expression showed that some neurons underwent oxidative stress around the ischemic boundary zone at 6h post-ischemia. Cilostazol treatment significantly reduced the percentage of 8-OHdG-positive neurons (65.8±33.5% with vehicle and 21.3±9.9% with cilostazol). Moreover, NADPH oxidase (NOX) 2-positive neurons were significantly reduced with cilostazol treatment. In contrast, immunohistochemistry results for phosphorylated cyclic-AMP response element binding protein (pCREB) showed that there were significantly more pCREB-positive neurons around the ischemic boundary zone of cilostazol-treated mice than in those of vehicle-treated mice at 24h post-ischemia. These results suggested that cilostazol might have multiple mechanisms of action to ameliorate ischemic tissue damage, by attenuating oxidative stress mediated by suppressing NOX2 expression by ischemic neurons and an anti-apoptotic effect mediated through the pCREB pathway.
  • Shichinohe Hideo, Kuroda Satoshi, Tan Chengbo, Abumiya Takeo, Nakayama Naoki, Kazumata Ken, Hokari Masaaki, Houkin Kiyohiro
    STROKE 46 2015年02月 [査読有り][通常論文]
  • Shimoda Yusuke, Nakayama Naoki, Hokari Masaaki, Abumiya Takeo, Shichinohe Hideo, Kazumata Ken, Houkin Kiyohiro
    STROKE 46 2015年02月 [査読有り][通常論文]
  • Kurisu Kota, Abumiya Takeo, Nakamura Hideki, Shimbo Daisuke, Kazumata Ken, Nakayama Naoki, Shichinohe Hideo, Hokari Masaaki, Osanai Toshiya, Shimizu Hiroshi, Houkin Kiyohiro
    STROKE 46 2015年02月 [査読有り][通常論文]
  • Abumiya Takeo, Fujima Noriyuki, Kudo Kohsuke, Ishi Yukitomo, Gekka Masayuki, Shichinohe Hideo, Kazumata Ken, Nakayama Naoki, Houkin Kiyohiro
    STROKE 46 2015年02月 [査読有り][通常論文]
  • Kazumata K, Tha KK, Narita H, Kusumi I, Shichinohe H, Ito M, Nakayama N, Houkin K
    Stroke 46 2 354 - 360 2015年02月 [査読有り][通常論文]
     
    BACKGROUND AND PURPOSE: The mechanisms underlying frontal lobe dysfunction in moyamoya disease (MMD) are unknown. We aimed to determine whether chronic ischemia induces subtle microstructural brain changes in adult MMD and evaluated the association of changes with neuropsychological performance. METHODS: MRI, including 3-dimensional T1-weighted imaging and diffusion tensor imaging, was performed in 23 adult patients with MMD and 23 age-matched controls and gray matter density and major diffusion tensor imaging indices were compared between them; any alterations in the patients were tested for associations with age, ischemic symptoms, hemodynamic compromise, and neuropsychological performance. RESULTS: Decrease in gray matter density, associated with hemodynamic compromise (P<0.05), was observed in the posterior cingulate cortex of patients with MMD. Widespread reduction in fractional anisotropy and increases in radial diffusivity and mean diffusivity in some areas were also observed in bilateral cerebral white matter. The fractional anisotropy (r=0.54; P<0.0001) and radial diffusivity (r=-0.41; P<0.01) of white matter significantly associated with gray matter density of the cingulate cortex. The mean fractional anisotropy of the white matter tracts of the lateral prefrontal, cingulate, and inferior parietal regions were significantly associated with processing speed, executive function/attention, and working memory. CONCLUSIONS: In adult MMD, there were more white matter abnormalities than gray matter changes. Disruption of white matter may play a pivotal role in the development of cognitive dysfunction.
  • Yamauchi T, Kuroda Y, Morita T, Shichinohe H, Houkin K, Dezawa M, Kuroda S
    PloS one 10 3 e0116009  3 2015年 [査読有り][通常論文]
     
    OBJECTIVE: Bone marrow stromal cells (BMSCs) are heterogeneous and their therapeutic effect is pleiotropic. Multilineage-differentiating stress enduring (Muse) cells are recently identified to comprise several percentages of BMSCs, being able to differentiate into triploblastic lineages including neuronal cells and act as tissue repair cells. This study was aimed to clarify how Muse and non-Muse cells in BMSCs contribute to functional recovery after ischemic stroke. METHODS: Human BMSCs were separated into stage specific embryonic antigen-3-positive Muse cells and -negative non-Muse cells. Immunodeficient mice were subjected to permanent middle cerebral artery occlusion and received transplantation of vehicle, Muse, non-Muse or BMSCs (2.5×104 cells) into the ipsilateral striatum 7 days later. RESULTS: Motor function recovery in BMSC and non-Muse groups became apparent at 21 days after transplantation, but reached the plateau thereafter. In Muse group, functional recovery was not observed for up to 28 days post-transplantation, but became apparent at 35 days post-transplantation. On immunohistochemistry, only Muse cells were integrated into peri-infarct cortex and differentiate into Tuj-1- and NeuN-expressing cells, while negligible number of BMSCs and non-Muse cells remained in the peri-infarct area at 42 days post-transplantation. CONCLUSIONS: These findings strongly suggest that Muse cells and non-Muse cells may contribute differently to tissue regeneration and functional recovery. Muse cells may be more responsible for replacement of the lost neurons through their integration into the peri-infarct cortex and spontaneous differentiation into neuronal marker-positive cells. Non-Muse cells do not remain in the host brain and may exhibit trophic effects rather than cell replacement.
  • Yamauchi T, Saito H, Ito M, Shichinohe H, Houkin K, Kuroda S
    Translational stroke research 5 6 701 - 710 6 2014年12月 [査読有り][通常論文]
     
    Autologous human bone marrow stromal cells (hBMSCs) should be expanded in the animal serum-free condition within clinically relevant periods in order to secure safe and effective cell therapy for ischemic stroke. This study was aimed to assess whether the hBMSCs enhance their proliferation capacity and provide beneficial effect in the infarct brain when cultured with platelet lysate (PL) and granulocyte-colony stimulating factor (G-CSF). The hBMSCs were cultured in the fetal calf serum (FCS)-, PL-, or PL/G-CSF-containing medium. Cell growth kinetics was analyzed. The hBMSCs-PL, hBMSC-PL/G-CSF, or vehicle was stereotactically transplanted into the ipsilateral striatum of the rats subjected to permanent middle cerebral artery occlusion 7 days after the insult. Motor function was assessed for 8 weeks, and the fate of transplanted hBMSCs was examined using immunohistochemistry. As the results, the hBMSCs-PL/G-CSF showed more enhanced proliferation than the hBMSCs-FCS and hBMSCs-PL. Transplantation of hBMSCs expanded with the PL- or PL/G-CSF-containing medium equally promoted functional recovery compared with the vehicle group. Histological analysis revealed that there were no significant differences in their migration, survival, and neural differentiation in the infarct brain between the hBMSCs-PL and hBMSCs-PL/G-CSF. These findings strongly suggest that the combination of PL and G-CSF may accelerate hBMSC expansion and serve safe cell therapy for patients with ischemic stroke at clinically relevant timing.
  • 脳卒中再生医療 脳梗塞をターゲットとした自己骨髄間質細胞の定位移植治療
    黒田 敏, 七戸 秀夫, 宝金 清博, 出澤 真理
    脳循環代謝 26 1 103 - 103 (一社)日本脳循環代謝学会 2014年11月
  • Tan C, Shichinohe H, Abumiya T, Nakayama N, Kazumata K, Hokari M, Hamauchi S, Houkin K
    Neuropathology : official journal of the Japanese Society of Neuropathology 35 3 197 - 208 2014年11月 [査読有り][通常論文]
     
    Recently, both basic and clinical studies demonstrated that bone marrow stromal cell (BMSC) transplantation therapy can promote functional recovery of patients with CNS disorders. A non-invasive method for cell tracking using MRI and superparamagnetic iron oxide (SPIO)-based labeling agents has been applied to elucidate the behavior of transplanted cells. However, the long-term safety of SPIO-labeled BMSCs still remains unclear. The aim of this study was to investigate the short-, middle- and long-term safety of the SPIO-labeled allogeneic BMSC transplantation. For this purpose, BMSCs were isolated from transgenic rats expressing green fluorescent protein (GFP) and were labeled with SPIO. The Na/K ATPase pump inhibitor ouabain or vehicle was stereotactically injected into the right striatum of wild-type rats to induce a lacunar lesion (n = 22). Seven days after the insult, either BMSCs or SPIO solution were stereotactically injected into the left striatum. A 7.0-Tesla MRI was performed to serially monitor the behavior of BMSCs in the host brain. The animals were sacrificed after 7 days (n = 7), 6 weeks (n = 6) or 10 months (n = 9) after the transplantation. MRI demonstrated that BMSCs migrated to the damage area through the corpus callosum. Histological analysis showed that activated microglia were present around the bolus of donor cells 7 days after the allogeneic cell transplantation, although an immunosuppressive drug was administered. The SPIO-labeled BMSCs resided and started to proliferate around the route of the cell transplantation. Within 6 weeks, large numbers of SPIO-labeled BMSCs reached the lacunar infarction area from the transplantation region through the corpus callosum. Some SPIO nanoparticles were phagocytized by microglia. After 10 months, the number of SPIO-positive cells was lower compared with the 7-day and 6-week groups. There was no tumorigenesis or severe injury observed in any of the animals. These findings suggest that BMSCs are safe after cell transplantation for the treatment of stroke.
  • Shichinohe H, Ishihara T, Takahashi K, Tanaka Y, Miyamoto M, Yamauchi T, Saito H, Takemoto H, Houkin K, Kuroda S
    Neurorehabilitation and neural repair 29 1 80 - 9 2014年03月 [査読有り][通常論文]
     
    Background. Transplantation of bone marrow stromal cells (BMSCs) may contribute to functional recovery after stroke. This study was designed to clarify their mechanisms, trophic effects of neurotrophic factors, and neural differentiation. Methods. Mouse neurons exposed to glutamate were cocultured with mouse BMSCs. Either neutralizing antibodies against brain-derived neurotrophic factor (BDNF) or nerve growth factor (NGF) or Trk inhibitor K252a was added to explore the mechanism of their protective effects. Fluorescence in situ hybridization (FISH) was used to assess BDNF or NGF mRNA expression in BMSCs. The mice were subjected to permanent focal ischemia, and 7 days later, either BMSCs or the vehicle was stereotactically transplanted into the ipsilateral striatum. The mouse brains were processed for FISH and immunostaining 2 or 4 weeks after transplantation. Results. BMSCs significantly ameliorated glutamate-induced neuronal death. Treatment with anti-BDNF antibody significantly reduced their protective effects. FISH analysis showed that the majority of BMSCs expressed BDNF and NGF mRNA in vitro. BMSC transplantation significantly improved the survival of neurons in peri-infarct areas. FISH analysis revealed that approximately half of BMSCs expressed BDNF and NGF mRNA 2 weeks after transplantation; however, the percentage of BDNF and NGF mRNA-positive cells decreased thereafter. Instead, the percentage of microtubule-associated protein 2-positive BMSCs gradually increased during 4 weeks after transplantation. Conclusions. These findings strongly suggest that BDNF may be a key factor underlying the trophic effects of BMSCs. BMSCs might exhibit the trophic effect in the early stage of cell therapy and the phenotypic change toward neural cells thereafter.
  • Shimbo D, Abumiya T, Shichinohe H, Nakayama N, Kazumata K, Houkin K
    Brain research 1554 59 - 66 Elsevier 2014年03月 [査読有り][通常論文]
     
    Despite successful revascularization, reperfusion after prolonged ischemia causes ischemia reperfusion (I/R) injury. Recruitment and activation of neutrophils is thought to be a key event causing I/R injury. We examined whether post-ischemic intra-arterial infusion of liposome-encapsulated hemoglobin (LEH), an artificial oxygen carrier without neutrophils, could reduce I/R injury in a rat transient middle cerebral artery occlusion (MCAO) model. Male Sprague-Dawley rats were subjected to 2-h MCAO and then were divided into three groups: (1) LEH group (n=7) infused with LEH (Hb concentration of 6 g/dl, 10 ml/kg/h) through the recanalized internal carotid artery for 2 h, (2) vehicle group (n=8) infused with saline (10 ml/kg/h) in the same manner as the LEH group, and (3) control group (n=9) subjected to recanalization only. After 24-h reperfusion, all rats were tested for neurological score and then sacrificed to examine infarct and edema volumes, myeloperoxidase (MPO) expression, matrix metalloproteinase-9 (MMP-9) expression and activity, and reactive oxygen species (ROS) production. Compared with the control group and the vehicle group, the LEH group showed a significantly better neurological score and significantly smaller infarct and edema volumes. MPO expression, MMP-9 expression and activity, and ROS production in the LEH group were also significantly lower than those in the control and vehicle groups. The results in the present study suggest that post-ischemic intra-arterial infusion of LEH can reduce I/R injury through reducing the effect of MMP-9, most likely produced by neutrophils. This therapeutic strategy may be a promising candidate to prevent I/R injury after thrombolysis and/or thromboectomy. (C) 2014 Elsevier B.V. All rights reserved.
  • Kazumata K, Shinbo D, Ito M, Shichinohe H, Kuroda S, Nakayama N, Houkin K
    Journal of stroke and cerebrovascular diseases : the official journal of National Stroke Association 23 6 1421 - 8 2014年02月 [査読有り][通常論文]
     
    BACKGROUND: Adult moyamoya disease (MMD) is known to have high incidence of cerebral microbleeds (cMBs); however, the clinical significance still remains unclear. We investigated the frequency of cMBs in a large number of patients and analyzed the patterns of MB distribution in association with the location of the hematoma and moyamoya vessels. METHODS: We studied 259 consecutive patients with MMD using prospectively collected database. One hundred ninety-one patients were eligible for the present study, and image analysis was performed retrospectively. The presence of cMBs and remains of hemorrhage were determined using gradient-echo T2*-weighted sequence (1.5 T). The development of moyamoya vessels was assessed on source images of time-of-flight magnetic resonance angiography. The analysis consists of descriptive assessment of the spatial relationship between cMB, remains of hemorrhage, and moyamoya vessels. Statistical analysis was performed to calculate relative risk ratio in the presence of cMBs in relation to the remains of hemorrhage (macrohematoma), age of onset, and the presence of concomitant moyamoya vessels. RESULTS: Thirty MBs were observed in 20 adult MMD patients (16.9%). MBs were located predominantly in the periventricular white matter (63.3%) followed by the basal ganglia/thalami (20%). Comparing the patients with cMBs from those without, hematoma was more frequently observed in patients with cMBs (odds ratio [OR] 4.29; 95% confidence interval [CI] 1.58-11.62; P=.0062). Patients with adult onset was more likely to demonstrate cMBs (14.4%) compared with the patients with pediatric onset (4.1%) (OR 3.93; 95% CI 1.11-13.91). Moyamoya vessels appeared in the lateral part of the trigon, and the periventricular white matter was significantly associated with the presence of cMBs (lateral part of the trigon; OR 3.29 [1.59-6.82], P=.0019, periventricle of the body of lateral ventricle; OR 2.40 [1.20-4.79], P=.0214, respectively). cMBs accompanied concomitant arteries in 23 (76.7%) lesions. The subependymal-leptomeningeal artery anastomosis was the most common pattern (n=20, 66.7%). CONCLUSIONS: Spatial relationship was demonstrated between the moyamoya vessels and perivascular hemosiderin deposition particularly around the subependymal-leptomeningeal anastomosis, suggesting the mechanism for the development of cMBs in MMD. Present study further supports previous findings that cMBs potentially serve as a marker for the bleeding-prone microangiopathy in MMD. The significance of the present study lies in selecting optimal surgical candidate for preventing future hemorrhage by the presence of the cMBs, whereas current surgical indication relying on the degree of ischemia frequently fails to detect patients with future hemorrhage.
  • Vuignier S, Ito M, Kurisu K, Kazumata K, Nakayama N, Shichinohe H, Shiga T, Kiss JZ, Tamaki N, Houkin K
    Acta neurochirurgica 155 11 2097 - 2104 2013年11月 [査読有り][通常論文]
     
    BACKGROUND: The prevalence of ivy sign on fluid-attenuated inversion recovery (FLAIR) imaging in patients with asymptomatic moyamoya disease is unclear. The aim of this study was to clarify the incidence of ivy sign in these patients, as well as the correlation between MRI and (15)O gas PET findings. METHODS: A retrospective analysis including 16 consecutive patients with asymptomatic moyamoya disease enrolled between 2001 and 2010 in a single center. FLAIR imaging at the initial visit was categorized as ivy sign present, negative, or equivocal. Hemodynamic and metabolic parameters were quantified in 11 of 16 patients by (15)O-gas positron emission tomography, and the relationship between ivy sign and (15)O-gas PET parameters was analyzed. Cerebrovascular events within the follow-up period (54 ± 28 months) were also examined. RESULTS: Five of 16 asymptomatic moyamoya patients (31.3 %) had positive ivy sign (6/30 hemispheres, 20 %). In (15)O-gas PET examinations, 18 % of 22 hemispheres had elevated oxygen extraction fraction values that were significantly associated with positive ivy sign. Of these 16 asymptomatic moyamoya patients, six patients (37.5 %) underwent combined surgical revascularization. In this series, no patients experienced ischemic stroke, but one had intraventricular bleeding 1 year after surgery. CONCLUSIONS: Ivy sign on FLAIR imaging is still not rare in patients with moyamoya disease, even when asymptomatic. Although optimal management is still under debate, ivy sign may be an indicator of misery perfusion, and patients with asymptomatic moyamoya disease and ivy sign on FLAIR imaging will benefit from more careful follow-up.
  • Saito H, Magota K, Zhao S, Kubo N, Kuge Y, Shichinohe H, Houkin K, Tamaki N, Kuroda S
    Stroke 44 10 2869 - 2874 2013年10月 [査読有り][通常論文]
     
    BACKGROUND AND PURPOSE: This study was aimed to assess whether (123)I-iomazenil (IMZ) single photon emission computed tomography can serially monitor the effects of bone marrow stromal cell (BMSC) transplantation on neuronal integrity in infarct brain of rats. METHODS: The BMSCs were harvested from green fluorescent protein-transgenic rats and were cultured. The rats were subjected to permanent middle cerebral artery occlusion. Their motor function was serially quantified throughout the experiments. The BMSCs or vehicle was stereotactically transplanted into the ipsilateral striatum at 7 days after the insult. Using small-animal single photon emission computed tomography/computed tomography apparatus, the (123)I-IMZ uptake was serially measured at 6 and 35 days after the insult. Finally, fluorescence immunohistochemistry was performed to evaluate the distribution of engrafted cells and their phenotypes. RESULTS: The distribution of (123)I-IMZ was markedly decreased in the ipsilateral neocortex at 6 days postischemia. The vehicle-transplanted animals did not show a significant change at 35 days postischemia. However, BMSC transplantation significantly improved the distribution of (123)I-IMZ in the peri-infarct neocortex as well as motor function. The engrafted BMSCs were densely distributed around cerebral infarct, and some of them expressed neuronal nuclear antigen and γ-aminobutyric acid type-A receptor. CONCLUSIONS: The present findings strongly suggest that the BMSCs may enhance functional recovery by improving the neuronal integrity in the peri-infarct area, when directly transplanted into the infarct brain at clinically relevant timing. (123)I-IMZ single photon emission computed tomography may be a promising modality to scientifically prove the beneficial effects of BMSC transplantation on the host brain in clinical situation.
  • Masahito Kawabori, Satoshi Kuroda, Masaki Ito, Hideo Shichinohe, Kiyohiro Houkin, Yuji Kuge, Nagara Tamaki
    NEUROPATHOLOGY 33 2 140 - 148 2013年04月 [査読有り][通常論文]
     
    Stereotactic transplantation of bone marrow stromal cells (BMSCs) enables efficient delivery to the infarct brain. This study was aimed to assess its optimal timing and cell dose for ischemic stroke. The BMSCs were harvested from the green fluorescent protein-transgenic rats and were labeled with quantum dots. The BMSCs (1x105 or 1x106) were stereotactically transplanted into the ipsilateral striatum of the rats subjected to permanent middle cerebral artery occlusion at 1 or 4 weeks post-ischemia. Motor function was serially assessed. Using in vivo near infrared (NIR) fluorescence imaging, the engrafted BMSCs were visualized at 3 weeks post-transplantation. Immunohistochemistry was performed to evaluate their fate. Functional recovery was significantly enhanced when both low and high doses of BMSCs were transplanted at 1 week post-ischemia, but such therapeutic effects were observed only when the high-dose BMSCs were transplanted at 4 weeks post-ischemia. Both optical imaging and immunohistochemistry revealed their better engraftment in the peri-infarct area when the high-dose BMSCs were transplanted at 1 or 4 weeks post-ischemia. These findings strongly suggest the importance of timing and cell dose to yield therapeutic effects of BMSC transplantation for ischemic stroke. Earlier transplantation requires a smaller number of donor cells for beneficial effects.
  • 七戸秀夫, 黒田敏, 宮本倫行, 山内朋裕, 斎藤久泰, 新保大輔, 数又研, 中山若樹, 寶金清博
    再生医療 12 237  2013年02月28日 [査読無し][通常論文]
  • Shimbo Daisuke, Abumiya Takeo, Shichinohe Hideo, Nakayama Naoki, Kazumata Ken, Houkin Kiyohiro, Ishizuka Takanobu
    STROKE 44 2 2013年02月 [査読有り][通常論文]
  • Ito Masaki, Houkin Kiyohiro, Niiya Yoshimasa, Uchino Haruto, Mabuchi Shoji, Nakayama Naoki, Kazumata Ken, Shichinohe Hideo, Sugiyama Taku, Ishii Nobuaki, Nomura Mikio
    STROKE 44 2 2013年02月 [査読有り][通常論文]
  • Michiyuki Miyamoto, Satoshi Kuroda, Songji Zhao, Keiichi Magota, Hideo Shichinohe, Kiyohiro Houkin, Yuji Kuge, Nagara Tamaki
    Journal of Nuclear Medicine 54 1 145 - 150 2013年01月 [査読有り][通常論文]
     
    This study aimed to assess whether 18F-FDG PET could serially monitor the beneficial effects of bone marrow stromal cells (BMSC) on cerebral glucose metabolism when transplanted into the infarct brain of rats. Methods: The BMSC from green fluorescent protein transgenic rats or vehicle was stereotactically transplanted into the ipsilateral striatum at 7 d after permanent middle cerebral artery occlusion of rats. Local glucose metabolism was semiquantitatively measured at 6 and 35 d after ischemia using 18F-FDG PET. Motor function was serially evaluated throughout the experiments. At 35 d after ischemia, immunohistochemistry was performed to evaluate the phenotype of BMSC and their effects on the expression of brain-type glucose transporters. Results: BMSC transplantation not only enhanced functional recovery but also promoted the recovery of glucose utilization in the periinfarct area when stereotactically transplanted at 1 wk after ischemia. The engrafted cells were widely distributed, and most expressed a neuron-specific protein, NeuN. BMSC transplantation also prevented the pathologic upregulation of glucose transporters in the periinfarct neocortex. Conclusion: The present findings strongly suggest that the BMSC may enhance functional recovery by promoting the recovery of local glucose metabolism in the periinfarct area when directly transplanted into the infarct brain at clinically relevant timing. The BMSC also inhibit the pathologic upregulation of brain-isoform glucose transporters type 1 and 3. 18F-FDG PET may be a valuable modality to scientifically prove the beneficial effects of BMSC transplantation on the host brain in clinical situations. COPYRIGHT © 2013 by the Society of Nuclear Medicine and Molecular Imaging, Inc.
  • Shichinohe H, Yamauchi T, Saito H, Houkin K, Kuroda S
    Acta neurobiologiae experimentalis 73 3 354 - 363 3 2013年 [査読有り][通常論文]
     
    This study was aimed to clarify if the bone marrow stromal cells (BMSCs) significantly improve functional outcome after lacunar stroke when stereotactically transplanted into the brain. Ouabain, a Na/K ATPase pump inhibitor, was stereotactically injected into the right striatum of Wistar rats. One week later, the superparamagnetic iron oxide (SPIO)-labeled rat BMSCs (n=7) or vehicle (n=8) were stereotactically transplanted into the left striatum. Using rotarod test, motor function was serially evaluated through the experiment. A 7.0-T MR apparatus was employed to serially monitor the migration of BMSCs in the host brain. Histological analysis was performed at 7 weeks after ouabain injection, i.e., 6 weeks after BMSC transplantation. Ouabain injection yielded the reproducible, focal lesion in the right striatum, causing continuous motor dysfunction throughout the experiment. BMSC transplantation significantly enhanced the recovery of motor function after ouabain injection. MR imaging demonstrated that the BMSCs aggressively migrated towards the lesion through the corpus callosum. Histological analysis supported the findings on MRI. The BMSCs significantly enhanced the neurogenesis in the subventricular zone (SVZ) on both sides. Some of them also expressed neuronal or astrocytic phenotypes in the neocortex, SVZ, corpus callosum, and peri-lesion area. These findings strongly suggest that the BMSCs may serve therapeutic impacts on lacunar stroke when stereotactically transplanted at clinically relevant timing.
  • 骨髄間質細胞の動脈内送達の治療効果(Therapeutic effects of intra-arterial delivery of bone marrow stromal cells)
    長内 俊也, 黒田 敏, 七戸 秀夫, 久下 裕司, 玉木 長良, 宝金 清博
    脳循環代謝 24 1 147 - 147 (一社)日本脳循環代謝学会 2012年11月
  • Masahito Kawabori, Satoshi Kuroda, Taku Sugiyama, Masaki Ito, Hideo Shichinohe, Kiyohiro Houkin, Yuji Kuge, Nagara Tamaki
    NEUROPATHOLOGY 32 3 217 - 226 2012年06月 [査読有り][通常論文]
     
    Recent studies have indicated that bone marrow stromal cells (BMSC) may improve neurological function when transplanted into an animal model of CNS disorders, including cerebral infarct. However, there are few studies that evaluate the therapeutic benefits of intracerebral and intravenous BMSC transplantation for cerebral infarct. This study was aimed to clarify the favorable route of cell delivery for cerebral infarct in rats. The rats were subjected to permanent middle cerebral artery occlusion. The BMSC were labeled with near infrared (NIR)-emitting quantum dots and were transplanted stereotactically (1 x 10(6) cells) or intravenously (3 x 10(6) cells) at 7 days after the insult. Using in vivo NIR fluorescence imaging technique, the behaviors of BMSC were serially visualized during 4 weeks after transplantation. Motor function was also assessed. Immunohistochemistry was performed to evaluate the fate of the engrafted BMSC. Intracerebral, but not intravenous, transplantation of BMSC significantly enhanced functional recovery. In vivo NIR fluorescence imaging could clearly visualize their migration toward the cerebral infarct during 4 weeks after transplantation in the intracerebral group, but not in the intravenous, group. The BMSC were widely distributed in the ischemic brain and some of them expressed neural cell markers in the intracerebral group, but not in the intravenous group. These findings strongly suggest that intravenous administration of BMSC has limited effectiveness at clinically relevant timing and intracerebral administration should be chosen for patients with ischemic stroke, although further studies would be warranted to establish the treatment protocol.
  • Kiyohiro Houkin, Masaki Ito, Taku Sugiyama, Hideo Shichinohe, Naoki Nakayama, Ken Kazumata, Satoshi Kuroda
    NEUROLOGIA MEDICO-CHIRURGICA 52 5 267 - 277 2012年05月 [査読有り][通常論文]
     
    Research on moyamoya disease has progressed remarkably in the past several decades. Indeed, many new facts concerning the epidemiology of the disease have been revealed and surgical treatments have been drastically improved. However, despite extensive research, the mechanism of moyamoya disease is still unknown. Consequently, the cardinal treatment of this disease has not yet been developed. For further clarification of its etiology, innovative studies are therefore indispensable. The aim of this paper is to review research on the pathogenesis of moyamoya disease to identify milestones in the direction of its true solution. Many hypotheses of the pathogenesis of moyamoya disease have been proposed in the past half century, including infection (viral and bacterial), autoimmune disorders, proteins abnormality, and gene abnormality. Some of these are now considered to be historical achievements. Others, however, can be still subjected to contemporary research. Currently, several genetic abnormalities are considered to offer the most probable hypothesis. In addition, interesting papers have been presented on the role of the endothelial progenitor cell on the pathogenesis of moyamoya disease. Intuitively, however, it appears that a single theory cannot always explain the pathogenesis of this disease adequately. In other words, the complex mechanism of several factors may comprehensively explain the formation of moyamoya disease. The "double hit hypothesis" is probably the best explanation for the complicated pathology and epidemiology of this disease.
  • Yasuhiro Chiba, Satoshi Kuroda, Toshiya Osanai, Hideo Shichinohe, Kiyohiro Houkin, Yoshinobu Iwasaki
    Neuropathology : official journal of the Japanese Society of Neuropathology 32 2 139 - 48 2012年04月 [査読有り][通常論文]
     
    This study was designed to clarify the effects of donor age on biological features of bone marrow stromal cells (BMSC), one of the candidates for cell transplantation therapy for CNS disorders, because many aged patients might require such therapy. This study was also aimed to test whether ex vivo treatments with granulocyte-colony stimulating factor (G-CSF) could modify biological properties of BMSC from aged donors and enhance its therapeutic effects in an animal model of traumatic brain injury. The BMSC were harvested from young (6-week-old) and aged (100-week-old) rats. The ageing significantly increased the senescence-associated β-galactosidase (SA-β-gal) activity of the cultured BMSC, and decreased their proliferative capacity and production of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF). As the next step, the rats were subjected to brain freezing injury by applying liquid nitrogen onto the neocortex through the thinned skull. The 6-week BMSC, 100-week BMSC, G-CSF-treated 100-week BMSC or vehicle were stereotactically injected into the ipsilateral striatum at 7 days post-injury. Transplantation of the 6-week BMSC, but not 100-week BMSC, significantly improved locomotor function. However, treatment of the 100-week BMSC with 0.1 µmol of G-CSF significantly improved their proliferation activity and growth factor production, and recovered therapeutic effects in the injured brain. In conclusion, donor age may largely determine biological aspects of BMSC. G-CSF may contribute to improve the outcome of BMSC transplantation therapy for CNS disorders in aged patients.
  • Shichinohe H, Kuroda S, Kudo K, Ito M, Kawabori M, Miyamoto M, Nakanishi M, Terae S, Houkin K
    Translational stroke research 3 1 99 - 106 2012年03月 [査読有り][通常論文]
     
    Recent studies have elucidated that transplantation of the bone marrow stromal cells (BMSC) has therapeutic potential for the central nervous system (CNS) disorders. However, no imaging modalities have been established to track the engrafted cells in the CNS in clinical situation. This study aimed to investigate the ability of magnetic resonance imaging (MRI) to visualize the BMSC labeled with superparamagnetic iron oxide (SPIO). The BMSC of mice were labeled with SPIO. Various numbers of the cells were injected into the agar phantom and were visualized using a 3.0-T MR apparatus. The SPIO-labeled cells were injected into the temperature-sensitive gelation polymer (TGP) hydrogel and were cultured for 7 days. They were also visualized just after the injection and at 7 days postinjection. After a 7-day culture, they were stained with Turnbull blue technique. T2-, T2*-, and susceptibility-weighted imaging could identify minimally 1,000 cells in the agar or TGP hydrogel, although it was difficult to quantify their number on MRI. All of these sequences could track the SPIO-labeled BMSC for at least 7 days when injected into the TGP. Turnbull blue staining revealed the survival and proliferation of the SPIO-labeled BMSC in the TGP for 7 days. The findings strongly suggest that the SPIO labeling may enable to track minimally 1,000 cells engrafted in the CNS on clinical MR apparatus. These data would be valuable to consider the application of imaging technique into cell transplantation therapy for CNS disorders.
  • Toshiya Osanai, Satoshi Kuroda, Taku Sugiyama, Masahito Kawabori, Masaki Ito, Hideo Shichinohe, Yuji Kuge, Kiyohiro Houkin, Nagara Tamaki, Yoshinobu Iwasaki
    Neurosurgery 70 2 435 - 44 2012年02月 [査読有り][通常論文]
     
    BACKGROUND: A noninvasive and effective route of cell delivery should be established to yield maximal therapeutic effects for central nervous system (CNS) disorders. OBJECTIVE: To elucidate whether intra-arterial delivery of bone marrow stromal cells (BMSCs) significantly promotes functional recovery in traumatic brain injury (TBI) in rats. METHODS: Rat BMSCs were transplanted through the ipsilateral internal carotid artery 7 days after the onset of cortical freezing injury. The BMSCs were labeled with fluorescent dye, and in vivo optical imaging was employed to monitor the behaviors of cells for 4 weeks after transplantation. Motor function was assessed for 4 weeks, and the transplanted BMSCs were examined using immunohistochemistry. RESULTS: In vivo optical imaging and histologic analysis clearly demonstrated that the intra-arterially injected BMSCs were engrafted during the first pass without systemic circulation, and the transplanted BMSCs started to migrate from the cerebral capillary bed to the injured CNS tissue within 3 hours. Intra-arterial BMSC transplantation significantly promoted functional recovery after cortical freezing injury. A subgroup of BMSCs expressed the phenotypes of neurons, astrocytes, and endothelial cells around the injured neocortex 4 weeks after transplantation. CONCLUSION: Intra-arterial transplantation may be a valuable option for prompt, noninvasive delivery of BMSCs to the injured CNS tissue, enhancing functional recovery after TBI. In vivo optical imaging may provide important information on the intracerebral behaviors of donor cells by noninvasive, serial visualization.
  • Kazumata Ken, Houkin Kiyohiro, Nakayama Naoki, Shichinohe Hideo, Osanai Toshiya, Shinpo Daisuke, Saito Hisayasu
    CEREBROVASCULAR DISEASES 34 32  2012年 [査読有り][通常論文]
  • Ito Masaki, Niiya Yoshimasa, Uchino Haruto, Itosaka Hiroyuki, Shichinohe Hideo, Kazumata Ken, Nakayama Naoki, Mabuchi Syoji, Houkin Kiyohiro
    CEREBROVASCULAR DISEASES 34 102  2012年 [査読有り][通常論文]
  • 新保大輔, 鐙谷武雄, 七戸秀夫, 中山若樹, 数又研, 宝金清博, 石塚隆伸
    脳循環代謝 24 1 178  2012年 [査読無し][通常論文]
  • 骨髄間質細胞移植は脳梗塞周囲の神経受容体機能を改善させる―123I‐iomazenil SPECTによる検討
    齋藤久泰, 黒田敏, 宮本倫行, 山内朋裕, 七戸秀夫, 趙松吉, 孫田惠一, 久保直樹, 久下裕司, 玉木長良, 寶金清博
    脳循環代謝 24 1 216  2012年 [査読無し][通常論文]
  • 骨髄間質細胞移植は脳梗塞後の局所糖代謝を改善する―小動物用PET/CTによる検討
    宮本倫行, 黒田敏, 趙松吉, 孫田恵一, 伊東雅基, 川堀真人, 丸一勝彦, 七戸秀夫, 宝金清博, 久下裕司, 玉木長良
    再生医療 11 191  2012年 [査読無し][通常論文]
  • 脳梗塞をターゲットとした骨髄間質細胞移植―新たな培養・移植・イメージング技術による展開
    黒田敏, 宮本倫行, 山内朋裕, 斉藤久泰, 伊東雅基, 川堀真人, 杉山拓, 千葉泰弘, 七戸秀夫, 宝金清博, 久下裕司, 趙松吉, 玉木長良
    再生医療 11 136  2012年 [査読無し][通常論文]
  • Ito M, Kuroda S, Sugiyama T, Shichinohe H, Takeda Y, Nishio M, Koike T, Houkin K
    Translational stroke research 2 3 294 - 306 2011年09月 [査読有り][通常論文]
     
    This study was aimed to test the hypothesis that human bone marrow stromal cells (hBMSC) expanded in fetal calf serum (FCS)-free, platelet lysate (PL)-containing medium would retain their capacity of migration, survival, and neural differentiation when transplanted into the infarct brain, using serial in vivo magnetic resonance imaging (MRI). Cell growth kinetic analysis revealed that hBMSC maintain their proliferative activity when cultured either in conventional FCS-containing medium or FCS-free, PL-containing medium. Subsequently, hBMSC were labeled with a superparamagnetic iron oxide agent and were stereotactically transplanted into the ipsilateral striatum of rats at 7 days after permanent middle cerebral artery occlusion. Serial MRI performed over 8 weeks revealed that they retain their migratory capacity towards the cerebral infarct. Moreover, double fluorescence immunohistochemistry also revealed that they preserve their capacity of differentiation into the neural cells in the peri-infarct area. The hBMSC expanded in the FCS-free, PL-containing medium retain their capacity of migration, survival, and differentiation when stereotactically transplanted into the infarct brain. The present findings strongly suggest the clinical utility of PL as a substitute to expand autologous hBMSC for cerebral infarct in the future.
  • Shichinohe H, Kuroda S, Sugiyama T, Ito M, Kawabori M, Nishio M, Takeda Y, Koike T, Houkin K
    Translational stroke research 2 3 307 - 315 2011年09月 [査読有り][通常論文]
     
    The donor cell culture in animal serum-free medium is quite important for the clinical application of cell transplantation therapy. This study was aimed to test the hypothesis that the human bone marrow stromal cells (hBMSC) expanded with fetal calf serum (FCS)-free, platelet lysate (PL)-containing medium retain their biological features favoring central nervous system regeneration. The hBMSC were cultured with 5% PL or 10% FCS. Their phenotypes were analyzed with flow cytometry, and their production of growth factors was quantified with enzyme-linked immunosorbent assay. Their capacity of neural differentiation was verified by immunocytochemistry. There was no significant difference in morphology and cell surface marker between the hBMSC-FCS and hBMSC-PL. Both of them were positive for CD44, CD90, CD105, and CD166 and were negative for CD34, CD45, and CD271. The production of human brain-derived neurotrophic factor, human hepatocyte growth factor, human β-nerve growth factor, and human platelet-derived growth factor-BB did not differ between the two groups, although the hBMSC-PL produced significantly more amount of TGF-β1 than the hBMSC-FCS. There was no significant difference in their in vitro differentiation into the neurons and astrocytes between the two groups. The hBMSC expanded with PL-containing medium retain their biological capacity of neural differentiation and neuroprotection. The PL may be a clinically valuable and safe substitute for FCS in expanding the hBMSC for cell therapy.
  • Taku Sugiyama, Satoshi Kuroda, Yukari Takeda, Mitsufumi Nishio, Masaki Ito, Hideo Shichinohe, Takao Koike
    NEUROSURGERY 68 6 1733 - 1742 2011年06月 [査読有り][通常論文]
     
    BACKGROUND: The donor cell culture in animal serum-free medium is important for the clinical application of cell transplantation therapy. Recently, human-derived platelet lysate (PL) gained interest as a substitute for fetal calf serum (FCS), but there are no studies that evaluate the validity of human bone marrow stromal cells (hBMSCs) expanded with PL-containing medium for central nervous system disorders. OBJECTIVE: To test the hypothesis that hBMSCs expanded with FCS-free, PL-containing medium can promote functional recovery after cerebral infarct. METHODS: hBMSCs were cultured in the FCS-or PL-containing medium. Cell-growth kinetics were analyzed. The vehicle or hBMSCs was stereotactically transplanted into the ipsilateral striatum of the rats subjected to permanent middle cerebral artery occlusion 7 days after the insult. Motor function was assessed for 8 weeks, and the fate of transplanted hBMSCs was examined using immunohistochemistry. RESULTS: There was no significant difference in hBMSC expansion between the 2 groups. Transplantation of hBMSCs expanded with the FCS-or PL-containing medium equally promoted functional recovery compared with the vehicle group. Histological analysis revealed that there were no significant differences in their migration, survival, and neural differentiation in the infarct brain between the 2 groups. CONCLUSION: hBMSCs expanded with PL-containing medium retained their capacity of migration, survival, and differentiation and significantly promoted functional recovery when stereotactically transplanted into the infarct brain. The PL may be a clinically valuable and safe substitute for FCS in expanding hBMSCs to regenerate the infarct brain.
  • Taku Sugiyama, Satoshi Kuroda, Toshiya Osanai, Hideo Shichinohe, Yuji Kuge, Masaki Ito, Masahito Kawabori, Yoshinobu Iwasaki
    NEUROSURGERY 68 4 1036 - 1047 2011年04月 [査読有り][通常論文]
     
    BACKGROUND: Noninvasive imaging techniques would be needed to validate the therapeutic benefits of cell transplantation therapy for central nervous system disorders. OBJECTIVE: To evaluate whether near-infrared (NIR)-emitting fluorescence tracer, quantum dots, would be useful to noninvasively visualize the bone marrow stromal cells (BMSC) transplanted into the infarct brain in living animals. METHODS: Rat BMSCs were labeled with QD800. In vitro and in vivo conditions to visualize NIR fluorescence were precisely optimized. The QD800-labeled BMSCs were stereotactically transplanted into the ipsilateral striatum of the rats subjected to permanent middle cerebral artery occlusion 7 days after the insult. Using the NIR fluorescence imaging technique, the behaviors of BMSCs were serially visualized during the 8 weeks after transplantation. RESULTS: NIR fluorescence imaging could noninvasively detect the NIR fluorescence emitted from the transplanted BMSCs engrafted in the peri-infarct neocortex through the scalp up to 8 weeks after transplantation. The intensity gradually increased and reached the peak at 4 weeks. The results were supported by the findings on ex vivo NIR fluorescence imaging and histological analysis. CONCLUSION: NIR fluorescence imaging is valuable in monitoring the behaviors of donor cells in the rodent brain. The results would allow new opportunities to develop noninvasive NIR fluorescence imaging as a modality to track the BMSCs transplanted into the brain.
  • 骨髄間質細胞移植は脳梗塞後の局所糖代謝を改善する―小動物用PET/CTによる検討
    宮本倫行, 黒田敏, 七戸秀夫, 伊東雅基, 川堀真人, 趙松吉, 孫田恵一, 久下裕司, 玉木長良, 宝金清博
    脳循環代謝 23 1 142  2011年 [査読無し][通常論文]
  • Yasuhiro Chiba, Satoshi Kuroda, Hideo Shichinohe, Masaaki Hokari, Toshiya Osanai, Katsuhiko Maruichi, Shunsuke Yano, Kazutoshi Hida, Yoshinobu Iwasaki
    Neuropathology : official journal of the Japanese Society of Neuropathology 30 3 241 - 50 2010年06月 [査読有り][通常論文]
     
    Transplanted bone marrow stromal cells (BMSC) promote functional recovery after spinal cord injury (SCI) through multiple mechanisms. A Rho kinase inhibitor, Fasudil also enhances axonal regeneration. This study was aimed to evaluate whether combination therapy of BMSC transplantation and Fasudil further enhances axonal regeneration and functional recovery in rats subjected to SCI. Fasudil or vehicle was injected for 2 weeks. BMSC or vehicle transplantation into the rostral site of SCI was performed at 7 days after injury. Neurological symptoms were assessed throughout the experiments. Fluoro-Ruby was injected into the dorsal funiculus of the rostral site of SCI at 63 days after injury. The fate of the transplanted BMSC was examined using immunohistochemistry. BMSC transplantation significantly increased the number of Fluoro-Ruby -labeled fibers of the dorsal corticospinal tracts at the caudal site of SCI, enhancing functional recovery of the hind limbs. Some of the engrafted BMSC were positive for Fluoro-Ruby, neuronal specific nuclear protein and microtubule-associated protein-2, suggesting that they acquired neuronal phenotypes and built synaptic connection with the host's neural circuits. Fasudil treatment also improved axonal continuity, but did not promote functional recovery. Combination therapy dramatically increased the number of Fluoro-Ruby-labeled fibers of the dorsal corticospinal tracts at the caudal site of SCI, but did not further boost the therapeutic effects on locomotor function by BMSC transplantation. The findings suggest that BMSC transplantation and Fasudil provide synergistic effects on axon regeneration after SCI, although further studies would be necessary to further enhance functional recovery.
  • Toshiya Osanai, Satoshi Kuroda, Hiroshi Yasuda, Yasuhiro Chiba, Katsuhiko Maruichi, Masaaki Hokari, Taku Sugiyama, Hideo Shichinohe, Yoshinobu Iwasaki
    Neurosurgery 66 6 1140 - 7 2010年06月 [査読有り][通常論文]
     
    OBJECTIVE: Recent studies have indicated that bone marrow stromal cells (BMSCs) have the potential to improve neurological function when transplanted into animal models of cerebral infarct. However, it is still undetermined how the BMSCs should be transplanted to obtain the most efficient therapeutic benefits safely. The aim of this study was to assess whether a thermoreversible gelation polymer (TGP) hydrogel acts as a noninvasive, valuable scaffold in BMSC transplantation for infarct brain. METHODS: The mice were subjected to permanent middle cerebral artery occlusion. Vehicle, BMSC suspension, or the BMSC-TGP construct was transplanted onto the ipsilateral intact neocortex at 7 days after the insult. Neurological symptoms were assessed throughout the experiments. The fate of the transplanted BMSC was examined 8 weeks after transplantation with immunohistochemistry. RESULTS: TGP hydrogel completely disappeared and provoked no inflammation in the host brain. Many transplanted cells were widely engrafted in the ipsilateral cerebrum, including the dorsal neocortex adjacent to the cerebral infarct in the BMSC-TGP construct-treated mice. Their number was significantly larger than in the BMSC-treated mice. The majority were positive for both NeuN and MAP2 and morphologically simulated the neurons. CONCLUSION: The findings suggest that surgical transplantation of tissue-engineered BMSCs onto the intact neocortex enhances the engraftment of donor cells around the cerebral infarct. These data may be useful in developing a noninvasive but efficient paradigm in neural tissue engineering. TGP hydrogel can be a promising candidate for valuable scaffolds in BMSC transplantation for central nervous system disorders because of its unique biochemical properties.
  • 中枢神経再生におけるバイオイメージング
    杉山 拓, 伊東 雅基, 七戸 秀夫, 黒田 敏
    再生医療 9 2 257 - 263 (株)メディカルレビュー社 2010年05月 [査読有り][通常論文]
     
    細胞移植を中心とした中枢神経再生は臨床応用の段階を迎えつつある。同一個体内で経時的に評価しうるバイオイメージングは、移植細胞のモニタリングやホスト側の神経機能評価を行う上で、不可欠なものであり、今後も更なる研究の展開が必要である。本稿では、これまでに展開されてきた移植細胞の標識、モニタリングの手法を、それぞれのモダリティの特徴とともに、最新の知見を織り交ぜて紹介する。(著者抄録)
  • Hideo Shichinohe, Satoshi Kuroda, Katsuhiko Maruichi, Toshiya Osanai, Taku Sugiyama, Yasuhiro Chiba, Ayumi Yamaguchi, Yoshinobu Iwasaki
    Neuropathology : official journal of the Japanese Society of Neuropathology 30 2 113 - 22 2010年04月 [査読有り][通常論文]
     
    There are few studies that denote whether bone marrow stromal cells (BMSC) and bone marrow-derived mononuclear cells (MNC) show the same therapeutic effects, when directly transplanted into the infarct brain. This study therefore aimed to compare their biological properties and behaviors in the infarct brain. Mouse BMSC were harvested and cultured. Mouse MNC were obtained through centrifugation techniques. Their cell markers were analyzed with FACS analysis. The MNC (10(6) cells; n = 10) or BMSC (2 x 10(5) cells; n = 10) were stereotactically transplanted into the ipsilateral striatum of the mice subjected to permanent middle cerebral artery occlusion at 7 days after the insult. Their survival, migration, and differentiation in the infarct brain were precisely analyzed using immunohistochemistry 4 weeks after transplantation. The MNC were positive for CD34, CD45, CD90, but were negative for Sca-1. The BMSC were positive for CD90 and Sca-1. The transplanted BMSC, but not MNC, extensively migrated into the peri-infarct area. Approximately 20% of the transplanted BMSC expressed a neuronal marker, NeuN in the infarct brain, although only 1.4% of the transplanted MNC expressed NeuN. These findings strongly suggest that there are large, biological differences between MNC and BMSC as cell sources of regenerative medicine for ischemic stroke.
  • Hiroshi Yasuda, Satoshi Kuroda, Hideo Shichinohe, Shintaro Kamei, Ryoichi Kawamura, Yoshinobu Iwasaki
    JOURNAL OF NEUROSURGERY 112 2 336 - 344 2010年02月 [査読有り][通常論文]
     
    Object. In this study the authors' aim was to assess whether fibrin matrix could act as an injectable, valuable scaffold in bone marrow stromal cell (BMSC) transplantation for injured CNS tissue. Methods. Both clotting time and 31) structure of fibrin matrix were analyzed with various concentrations of fibrinogen and CaCl(2). The BMSCs were harvested from green fluorescent protein-transgenic mice and cultured. A cortical lesion was produced in rats by application of a very cold rod to the right cerebral hemisphere. The BMSCs, fibrin matrix, or BMSC-fibrin matrix complex was transplanted into the lesion though a small bur hole 7 days after the insult. Using immunohistochemical analysis, the authors evaluated the survival, migration, and differentiation of the transplanted cells 4 weeks after transplantation. Results. Based on in vitro observations, the concentrations of fibrinogen and CaCl(2) were fixed at 2.5 mg/ml and 2 mu M in animal experiments, respectively. Fibrin matrix almost completely disappeared 4 weeks after transplantation. However, immunohistochemical analysis revealed that fibrin matrix exclusively enhanced the retention of the transplanted cells within the lesion. migration toward the lesion boundary zone, and differentiation into the neurons and perivascular cells. Conclusions. Injectable fibrin matrix enhanced the Survival, migration, and differentiation of the BMSCs transplanted into the cortical lesion in rats. The authors believe that it is one of the promising candidates for a potential, minimally invasive scaffold for CNS disorders. The present findings strongly suggest that such a strategy of tissue engineering could be a therapeutic option for CNS regeneration in patients with CNS injuries. (DOI: 10.3171/10.3171/2009.2.JNS08495)
  • Hideo Shichinohe, Satoshi Kuroda, Taku Sugiyama, Masaki Ito, Masahito Kawabori
    DEMENTIA AND GERIATRIC COGNITIVE DISORDERS 30 4 293 - 301 2010年 [査読有り][通常論文]
     
    Aims: This study was aimed to elucidate if bone marrow stromal cells (BMSC) could ameliorate cognitive dysfunction due to chronic cerebral ischemia when transplanted into the brain. Methods: The BMSC were harvested from green fluorescence protein (GFP)-expressing mice. Wistar rats were subjected to bilateral common carotid artery (CCA) ligation. The BMSC (4 x 10(5) cells) or vehicle were stereotactically injected into the right striatum 24 h after the insult. Cognitive function was evaluated with the Morris water maze task after 3 and 5 weeks. Histological analysis was performed after 6 weeks. Results: Cognitive function was significantly impaired in the vehicle-transplanted animals, when compared with the non-CCA-ligation animals. BMSC transplantation significantly improved it. The BMSC were widely distributed in the ischemic brain, including the neocortex, white matter and hippocampus, and some of them expressed the phenotypes of neurons, astrocytes and endothelium. They also significantly ameliorated white matter damage. Conclusions: These findings strongly suggest that the BMSC may have the potential to attenuate white matter injury and improve cognitive dysfunction due to chronic cerebral ischemia. The present results would shed light on the potential of a novel strategy, cell therapy against ischemia-related cognitive dysfunction. Copyright (C) 2010 S. Karger AG, Basel
  • Katsuhiko Maruichi, Satoshi Kuroda, Yasuhiro Chiba, Masaaki Hokari, Hideo Shichinohe, Kazutoshi Hida, Yoshinobu Iwasaki
    NEUROPATHOLOGY 29 4 422 - 432 2009年08月 [査読有り][通常論文]
     
    Diffuse axonal injury (DAI) often leads to persistent cognitive dysfunction in spite of the lack of gross lesions on MRI. Therefore, this study was aimed to evaluate whether transplanted bone marrow stromal cells (BMSC) can improve DAI-induced cognitive dysfunction or not. The rats were subjected to impact acceleration head injury, using a pneumatic high-velocity impactor. The BMSC were harvested from the mice and were cultured. The BMSC (4.0 x 10(5) cells) or vehicle were stereotactically transplanted into the right striatum at 10 days post-injury. Cognitive function analysis was repeated at 1, 2, and 4 weeks post-injury, using the Morris water maze test. Histological analysis was performed at 2, 8 and 20 weeks post-injury, using double fluorescence immunohistochemistry. Transplanted BMSC were widely distributed in the injured brain and gradually acquired the phenotypes of neurons and astrocytes over 20 weeks. In addition, they significantly improved DAI-induced cognitive dysfunction as early as 2 weeks post-injury, although their processes of neuronal differentiation were not completed at this time point. The findings suggest that the engrafted BMSC may exhibit this early beneficial effect on cognitive function by producing neuroprotective or neurotrophic factors. In conclusion, direct transplantation of BMSC may serve as a novel therapeutic strategy to enhance the recovery from DAI-induced cognitive impairment.
  • Hiroyuki Itosaka, Satoshi Kuroda, Hideo Shichinohe, Hiroshi Yasuda, Shunsuke Yano, Shintaro Kamei, Ryoichi Kawamura, Kazutoshi Hida, Yoshinobu Iwasaki
    NEUROPATHOLOGY 29 3 248 - 257 2009年06月 [査読有り][通常論文]
     
    Recent basic experiments have strongly suggested that cell transplantation therapy may promote functional recovery in patients with spinal cord injury (SCI). However, a safe and efficient transplantation technique still remains undetermined. This study, therefore, was aimed to clarify whether fibrin matrix could be a useful scaffold in bone marrow stromal cell (BMSC) transplantation for the injured spinal cord. To clarify the issue, three-dimensional structure of fibrin matrix was assessed and the green fluorescent protein (GFP)-expressing BMSC were cultured in fibrin matrix. The rats were subjected to spinal cord hemisection at T8 level, and the vehicle, BMSC or BMSC-fibrin matrix construct was implanted into the cavity. Neurologic function was serially evaluated. Using immunohistochemistry, we evaluated the survival, migration and differentiation of the transplanted cells at 4 weeks after transplantation. In the initial in vitro study, the BMSC could survive in fibrin matrix for 2 weeks. The animals treated with the BMSC-fibrin matrix construct showed significantly more pronounced recovery of neurologic function than vehicle- or BMSC-treated animals. Fibrin scaffold markedly improved the survival and migration of the transplanted cells. There was no significant difference in the percentage of cells doubly positive for GFP and microtubule-associated protein 2 between the animals treated with BMSC-fibrin matrix construct and those treated with BMSC, but a certain subpopulation of GFP-positive cells morphologically simulated the neurons in the animals treated with BMSC-fibrin matrix construct. These findings strongly suggest that fibrin matrix may be one of the promising candidates for a potential, minimally invasive scaffold for injured spinal cord, and that such strategy of tissue engineering could be a hopeful option in regeneration therapy for patients with SCI.
  • Yasuhiro Chiba, Satoshi Kuroda, Katsuhiko Maruichi, Toshiya Osanai, Masaaki Hokari, Shunsuke Yano, Hideo Shichinohe, Kazutoshi Hida, Yoshinobu Iwasaki
    Neurosurgery 64 5 991 - 9 2009年05月 [査読有り][通常論文]
     
    OBJECTIVE: Recent studies have indicated that bone marrow stromal cells (BMSCs) have the potential to improve neurological function when transplanted into animal models of spinal cord injury (SCI). However, it is still unclear how the transplanted BMSCs promote functional recovery after SCI. In this study, therefore, we evaluated how the transplanted BMSCs restore the function of the dorsal corticospinal tracts in the injured spinal cord. METHODS: The rats were subjected to incomplete SCI by means of a pneumatic impact device. BMSC or vehicle transplantation into the rostral site of SCI was performed at 7 days after injury. Neurological symptoms were assessed throughout the experiments. Fluoro-Ruby was injected into the dorsal funiculus of the rostral site of SCI at 63 days after injury. The fate of the transplanted BMSCs was examined using immunohistochemistry. RESULTS: BMSC transplantation significantly enhanced functional recovery of the hind limbs. The number of Fluoro-Ruby-labeled fibers of the dorsal corticospinal tracts at the caudal site of SCI was significantly higher in the BMSC-transplanted animals than in the vehicle-transplanted animals. Some of the engrafted BMSCs were positive for Fluoro-Ruby, NeuN, and MAP2 in the gray matter, suggesting that they acquired neuronal phenotypes and built synaptic connection with the host's neural circuits. Others in the white matter morphologically simulated the astrocytes and were also positive for glial fibrillary acidic protein. CONCLUSION: The findings suggest that the transplanted BMSCs acquire neural cell phenotypes around the injury site and contribute to rebuild the neural circuits, including the corticospinal tract, promoting functional recovery of the hind limbs.
  • Katsuhiko Maruichi, Satoshi Kuroda, Yasuhiro Chiba, Masaaki Hokari, Hideo Shichinohe, Kazutoshi Hida, Yoshinobu Iwasaki
    NEUROPATHOLOGY 29 2 132 - 139 2009年04月 [査読有り][通常論文]
     
    Diffuse axonal injury (DAI) plays a major role in the development of cognitive dysfunction, emotional difficulties and behavioral disturbances in patients following closed head injury, even when they have no definite abnormalities on conventional MRI. This study aimed to develop a highly controlled and reproducible model for DAI that simulates post-traumatic cognitive dysfunction in humans. Sprague-Dawley (SD) rats were subjected to impact acceleration head injury, using a pneumatic impact targeted to a steel disc centered onto their skull. The severity of injury was graded as three levels by adjusting the driving pressure at 60, 70 or 80 pounds per square inch. In vivo MRI was obtained 2 days post-injury. Cognitive function was evaluated using the Morris water maze at 1 and 2 weeks post-injury. HE staining and immunohistochemistry were performed to assess neuronal and axonal damages after 2 weeks. MRI demonstrated that this model induced no gross structural modification in the brain. The degree and duration of cognitive dysfunction were dependent on the force of impact. Histological analysis revealed the force-dependent damage of the neurons and microtubule-associated protein 2-positive axons in the neocortex. Hippocampal damage was much less pronounced and was not linked to cognitive dysfunction. This is the first report that precisely evaluates the threshold of impact energy to lead to neocortical damage and cognitive dysfunction in rodents. This model would be suitable for clarifying the complex mechanisms of post-traumatic brain damage and testing novel therapeutic approaches against post-traumatic cognitive dysfunction due to diffuse axonal damage.
  • Takeshi Aoyama, Kazutoshi Hida, Satoshi Kuroda, Toshitaka Seki, Shunsuke Yano, Hideo Shichinohe, Yoshinobu Iwasaki
    NEUROLOGIA MEDICO-CHIRURGICA 48 12 539 - 545 2008年12月 [査読有り][通常論文]
     
    The present study evaluated the effect of the free radical scavenger edaravone on lesion volume and neurological dysfunction after spinal cord injury (SCI) in mice, and investigated its protective effects on superoxide generation. Female C57BL/6 mice were subjected to SCI using a pneumatic impact device and were treated with 3 mg/kg of edaravone or vehicle 30 minutes before the insult. Motor functions were quantitatively evaluated. Lesion volume was assessed by Dohrmann's two-cone method after one week. In situ detection of superoxide in the injured cord was carried out using the superoxide-sensitive dye dihydroethidium (DHE) staining technique. Pretreatment with edaravone significantly improved motor dysfunction and reduced the lesion volume to about 63% of the control (p < 0.05). Semi-quantitative measurements of red fluorescence emitted from DHE revealed that the superoxide concentration increased in the lesion periphery at 1 and 3 hours after the insult, and that pretreatment with edaravone significantly inhibited the increase of superoxide concentration in the lesion periphery at both time points (p < 0.0001). Double staining with DHE and monoclonal antibody against MAP2 showed that most cells positive for DHE were also positive for MAP2. These findings suggest that edaravone ameliorates tissue damage by scavenging reactive oxygen species, especially in the neurons, after SCI.
  • Hideo Shichinohe, Satoshi Kuroda, Sachiko Tsuji, Satoshi Yamaguchi, Shunsuke Yano, Jang-Bo Lee, Hiroyuki Kobayashi, Seiji Kikuchi, Kazutoshi Hida, Yoshinobu Iwasaki
    NEUROREHABILITATION AND NEURAL REPAIR 22 5 447 - 457 2008年09月 [査読有り][通常論文]
     
    Objective. Recent reports have indicated that bone marrow stromal cells (BMSCs) have the potential to improve neurological function when transplanted into models of central nervous System (CNS) disorders, including traumatic spinal cord injury. In this study, the authors aimed to clarify the underlying mechanism through which BMSCs supported CNS regeneration in the spinal cord. Methods. The authors topically applied mouse BMSCs expressing green fluorescence protein (0.4-4 x 10(4) cells) on the organotypic spinal cord slice culture prepared front 6-day-old rat pups (n = 17). They were co-cultured for 3 weeks after the Slice Culture started, and the behavior of the applied BMSCs was serially observed using a fluorescence bioimaging technique. The authors completed a histological analysis at the end of the co-cultures and evaluated the profiles of the Cultured BMSCs using microarray and immunocytochemistry techniques. Results. The fluorescence bioimaging showed that the BMSCs Survived and made a cluster on the slice during the experiments. They also induced a morphological change in the slice within 48 hours of co-culture. Immunohistochemistry analysis showed that the BMSCs promoted a marked neurite extension toward their Cluster and some of the BMSCs expressed Tuj-1, an early neuronal marker. Analysis by microarray and immunocytochemistry revealed that BMSCs highly expressed the matrix metalloproteinases (MMPs), stromal cell-derived factor-1, and its specific receptor CXCR4. Conclusions. These findings Suggest that the donor BMSCs can support CNS regeneration due to their acquisition of a suitable environment for differentiation and promotion of neurite extension via MMPs and chemokines.
  • Nobutaka Horie, Arme-Lise Maag, Scott A. Hamilton, Hideo Shichinohe, Tonya M. Bliss, Gary K. Steinberg
    JOURNAL OF NEUROSCIENCE METHODS 173 2 286 - 290 2008年08月 [査読有り][通常論文]
     
    intracerebral injection of the vasoconstrictor peptide, endothelin-1 (ET-1), has been used as a method to induce focal ischemia in rats. The relative technical simplicity of this model makes it attractive for use in mice. However, the effect of ETA on mouse brains has not been firmly established. In this study, we determined the ability of ET-1 to induce focal cerebral ischemia in four different mouse strains (CD1, C57/BL6, NOD/SCID, and FVB). In contrast to rats, intracerebral injection of ET-1 did not produce a lesion in any mouse strain tested. A combination of ET-1 injection with either CCA occlusion or N(G)-nitro-L-arginine methyl ester (L-NAME) injection produced only a small infarct and its size was strain-dependent. A triple combination of CCA occlusion with co-injection of ETA and L-NAME produced a lesion in all mouse strains tested, and this resulted in a significant motor deficit. However, lesion size was still relatively small and strain-dependent. This study shows that ET-1 has a much less potent effect for producing an infarct in mice than rats. (c) 2008 Elsevier B.V. All rights reserved.
  • Tomoko Matoba, Yasuko Orba, Tadaki Suzuki, Yoshinori Makino, Hideo Shichinohe, Satoshi Kuroda, Takahiro Ochiya, Hiroshi Itoh, Shinya Tanaka, Kazuo Nagashima, Hirofumi Sawa
    NEUROPATHOLOGY 28 3 286 - 294 2008年06月 [査読有り][通常論文]
     
    JC virus (JCV) is the etiological agent of the demyelinating disease progressive multifocal leukoencephalopathy (PML). Because JCV has a very narrow host range, it has been difficult to develop an animal model of JCV infection; as a result, no effective therapy for PML has been established. In this study, we have tried to create an animal model that replaces an in vivo JCV infection. As a result, we have obtained a stable persistence of JCV-infected human cells in the mouse brain by inoculating the virus-infected cells into the nude mice brains. In this model, the JCV-infected cells were well preserved in the nude mouse brains for 2 weeks. We then treated JCV-injected brains with an siRNA against the JCV agnoprotein that is known to be an effective inhibitor of JCV infection in vitro. A highly purified type I collagen, atelocollagen, was used as a carrier for the siRNA. The siRNA inhibited the expression of JCV protein in inoculated JCV-infected cells in the mouse brain, compared to the medium containing only atelocollagen used as a placebo. Thus, the combination of siRNA and atelocollagen might be a candidate therapeutic agent for the treatment of JCV infection.
  • Masaaki Hokari, Satoshi Kuroda, Hideo Shichinohe, Shunsuke Yano, Kazutoshi Hida, Yoshinobu Iwasaki
    JOURNAL OF NEUROSCIENCE RESEARCH 86 5 1024 - 1035 2008年04月 [査読有り][通常論文]
     
    A surprising shortage of information surrounds the mechanism by which bone marrow stromal cells (BMSC) restore lost neurologic functions when transplanted into the damaged central nervous system. To clarify the issue, the BMSC were cocultured with the neurons using two paradigms: the cell-mixing coculture technique and three-dimensional coculture technique. The green fluorescent protein (GFP)-expressing BMSC were cocultured with the PKH-26-labelled neurons, using cell mixing coculture technique. GFP-positive, PKH-26-negative cells morphologically simulated the neurons and significantly increased the expression of AP-2, Tuj-1, nestin, and GFAP GFP/nestin-positive, PKH-26-negative cells increased from 13.6% +/- 6.7% to 32.1 % +/- 15.5% over 7 days of coculture. They further enhanced Tuj-1 expression when cocultured with neurons exposed to 100 mu M of glutamate for 10 min. About 20-30% of GFP-positive cells became positive for PKH-26 through coculture with the neurons, but the doubly positive cells did not increase when cocultured with glutamate-exposed neurons. Alternatively, the BMSC significantly ameliorated glutamate-induced neuronal damage when cocultured with the three-dimensional coculture technique. The protective effect was more prominent when coculture was started prior to glutamate exposure than when coculture was started just after glutamate exposure. ELISA analysis revealed that the BMSC physiologically produce NGF, BDNF, SDF-1 alpha, HGF, TGF beta-1, and IGF-1 and significantly enhanced the production of NGF and BDNF when cocultured with glutamate-exposed neurons. These findings strongly suggest that the BMSC may protect and repair the damaged neurons through multiple mechanisms, including transdifferentiation, cell fusion, and production of growth factors. (C) 2007 Wiley-Liss, Inc.
  • Hideo Shichinohe, Satoshi Kuroda, Shunsuke Yano, Kazutoshi Hida, Yoshinobu Iwasaki
    BRAIN RESEARCH 1183 138 - 147 2007年12月 [査読有り][通常論文]
     
    Recent studies have indicated that bone marrow stromal cells (BMSC) have the potential to improve neurological function when transplanted into animal models of cerebral infarction. However, it is still obscure how the transplanted BMSC restore the lost neurological function. in this study, therefore, we aimed to elucidate the role of stromal cell-derived factor-1 (SDF-1) and its specific receptor, CXCR4, in BMSC transplantation into the brain subjected to cerebral infarction. The BMSC were harvested from the wild type (WT) and CXCR4-knockout (CXCR4-KO) mice and were cultured. The mice were subjected to permanent middle cerebral artery occlusion. The WT or CXCR4-KO BMSC was injected into the ipsilateral striatum 7 days after the insult. Motor function of the animals was serially evaluated, using a rotarod treadmill. Using fluorescence immunohistochemistry, we evaluated the distribution and phenotype of the transplanted cells 4 weeks after transplantation. Recovery of motor function in the WT BMSC-transplanted mice was more pronounced than in the CXCR4-KO-transplanted mice and the vehicle-treated ones. SDF-1 was extensively expressed in peri-infarct area. In the WT BMSC-transplanted mice, the transplanted cells were extensively distributed in the ipsilateral hemisphere, and many of them migrated towards the peri-infarct area and expressed the proteins specific for neurons and astrocytes, although these findings were not observed in the CXCR4-KO-transplanted mice. The results suggest that the SDF-1/CXCR4 system may play a critical role in the survival, proliferation and migration of the transplanted BMSC and contribute to recovery of neurological function. (c) 2007 Elsevier B.V. All rights reserved.
  • Shunsuke Yano, Satoshi Kuroda, Hideo Shichinohe, Toshitaka Seki, Takako Ohnishi, Hiroshi Tamagami, Kazutoshi Hida, Yoshinobu Iwasaki
    JOURNAL OF NEUROTRAUMA 23 11 1682 - 1692 2006年11月 [査読有り][通常論文]
     
    A surprising shortage of information surrounds the mechanisms by which bone marrow stromal cells (BMSC) restore lost neurologic functions when transplanted into the damaged central nervous system. In the present study, we sought to elucidate whether BMSCs express the neuron-specific gamma-aminobutyric acid (GABA) receptor when transplanted into injured spinal cord. To examine this, we harvested and cultured rat femoral BMSCs. We then subjected Sprague-Dawley rats to thoracic spinal cord injury (SCI) with a pneumatic impact device. Fluorescence-labeled BMSCs (n = 7) were transplanted stereotactically or the vehicle in which these cells were cultured (n = 4) was introduced stereotactically into the rostral site of SCI at 7 days after injury. We evaluated GABA receptor function by measuring the binding potential for I-125-iomazenil (I-121-IMZ) through in vitro autoradiography at 4 weeks after BMSC transplantation and simultaneously examined the fate of the transplanted BMSCs by immunocytochemistry. We found that the transplanted BMSC migrated toward the core of the injury and were densely distributed in the marginal region at 4 weeks after transplantation. BMSC transplantation significantly increased the binding potential for I-121-IMZ (p = 0.0376) and increased the number of GABA receptor-positive cells (P = 0.0077) in the marginal region of the injury site. Some of the transplanted BMSCs were positive for microtubule-associated protein-2 and the alpha 1 subunit of GABA(A) receptor in the region of injury. These findings suggest that BMSCs have the potential to support the survival of neurons in the marginal region of SCI and can partly differentiate into neurons, regenerating spinal cord tissue at the site of injury.
  • Yoshimasa Niiya, Takeo Abumiya, Hideo Shichinohe, Satoshi Kuroda, Seiji Kikuchi, Masahiro Ieko, Sho-ichi Yamagishi, Masayoshi Takeuchi, Takashi Sato, Yoshinobu Iwasaki
    BRAIN RESEARCH 1108 179 - 187 2006年09月 [査読有り][通常論文]
     
    There is accumulating evidence that advanced glycation end products (AGES) are relevant to the formation of vascular complications in diabetes mellitus. The aim of this study was to investigate whether AGES have a significant effect on tissue factor (TF) expression in brain microvascular endothelial cells compared with that in other arterial endothelial cells. Cultured bovine brain microvascular endothelial cells (BBMECs) and aortic endothelial cells (BAECs) were incubated in medium containing glyceraldehyde-derived AGE (glycer-AGE). TF mRNA expression, protein expression, and activity were measured at multiple time points after glycer-AGE incubation. Participation of reactive oxygen species (ROS) in the effect of glycer-AGE on TF expression was investigated by treatment with a free radical scavenger, edaravone, and intracellular ROS measurements with dihydroethidium (DHE). Basic TF mRNA expression was greater in BBMECs than in BAECs. Glycer-AGE significantly upregulated TF mRNA expression in both cells, and the upregulation was more prominent in BBMECs than in BAECs. TF protein expression and activity were also upregulated with a pattern of being greater in BBMECs than in BAECs. Edaravone significantly attenuated the AGE-induced upregulation of TF mRNA expression, protein expression, and activity. Intracellular ROS levels measured with DHE-stained fluorescent intensity were significantly upregulated by glycer-AGE with a pattern of being greater in BBMECs than in BAECs. AGE-induced ROS upregulation was attenuated by edaravone like AGE-induced TF upregulation. These results suggest that brain microvascular endothelial cells are more susceptible to AGE-induced TF upregulation than aortic endothelial cells, and that this susceptibility is associated with levels of intracellular ROS. (c) 2006 Elsevier B.V. All rights reserved.
  • S Yamaguchi, S Kuroda, H Kobayashi, H Shichinohe, S Yano, K Hida, K Shinpo, S Kikuchi, Y Iwasaki
    BRAIN RESEARCH 1087 15 - 27 2006年05月 [査読有り][通常論文]
     
    Bone marrow stromal cells (BMSC) have been anticipated as a donor for cell type for transplantation therapy in various neurological disorders. However, their neurogenic capacity still remains undetermined. In this study, we aimed to clarify whether in vitro chemical treatment promotes their neuronal differentiation on the level of gene expression. Mice BMSC were cultured with medium supplemented with DMSO, retinoic acid, and basic fibroblast growth factor, and their morphology and expression of neuronal markers were evaluated. Subsequently, using microarray and RT-PCR techniques, the treatment-induced changes in the gene expression profile were analyzed. After exposure to the medium, the BMSC simulated a neuron-like appearance and increased their immunoreactivity for nestin and Tuj-1. Microarray analysis revealed that the BMSC per se express the multilineage cellular genes, including those associated with the neuron. Chemical treatment significantly decreased the expression of genes related to mesenchymal cells and increased the expression of 5 neuron-associated genes. Microarray and RT-PCR analyses also demonstrated that the BMSC express the genes for several growth factors including NGF-beta and BDNF, indicating their therapeutic role in protecting the injured central nervous system. The present results suggest that at least a certain subpopulation of the BMSC have the potential to alter their gene expression profile in response to the surrounding environment and may possibly protect the host tissue by secreting neuroprotective factors. (c) 2006 Elsevier B.V. All rights reserved.
  • Hideo Shichinohe, Satoshi Kuroda, Shunsuke Yano, Takako Ohnishi, Hiroshi Tamagami, Kazutoshi Hida, Yoshinobu Iwasaki
    JOURNAL OF NUCLEAR MEDICINE 47 3 486 - 491 2006年03月 [査読有り][通常論文]
     
    Recent studies have indicated that bone marrow stromal cells (BMSC) have the potential to improve neurologic function when transplanted into animal models of central nervous system disorders. However, how the transplanted BMSC restore the lost neurologic function is not clear. In the present study, therefore, we aimed to elucidate whether BMSC express the neuron-specific gamma-aminobutyric acid (GABA) receptor when transplanted into brain that has been subjected to cerebral infarction. Methods: The BMSC were harvested from green fluorescent protein-transgenic mice and were cultured. The mice were subjected to permanent middle cerebral artery occlusion. The BMSC or vehicle was transplanted into the ipsilateral striatum 7 d after the insult. Using autoradiography and fluorescence immunohis- to chemistry, we evaluated the binding of I-125-iomazenil and the expression of GABA receptor protein in and around the cerebral infarct 4 wk after transplantation. Results: Binding of I-125-iomazenil was significantly higher in the periinfarct neocortex in the BMSC-transplanted animals than in the vehicle-transplanted animals. Likewise, the number of the GABAA receptor-positive cells was significantly higher in the periinfarct neocortex in the BMSC-transplanted animals than in the vehicle-transplanted animals. A certain subpopulation of the transplanted BMSC expressed a neuron-specific marker, microtubule-associated protein 2, and the marker protein specific for GABAA receptor in the periinfarct area. Conclusion: These findings suggest that BMSC may contribute to neural tissue regeneration through migrating toward the periinfarct area and acquiring the neuron-specific receptor function.
  • 鐙谷武雄, 七戸秀夫, 中山若樹, 黒田敏, 寺坂俊介, 岩崎喜信
    脳卒中 28 1 194  2006年03月01日 [査読無し][通常論文]
  • S Yano, S Kuroda, H Shichinohe, K Hida, Y Iwasaki
    BRAIN RESEARCH 1065 1-2 60 - 67 2005年12月 [査読有り][通常論文]
     
    The present study was aimed to clarify the proliferation capacity of the bone marrow stromal cells (BMSC) transplanted into the brain. The BMSC were harvested from green fluorescence protein (GFP)-transgenic mice, grown to the confluency and passed three times. They were labeled by co-culture with Ferucarbotran, a superparamagnetic iron oxide (SPIO) agent. The proportions of the SPIO-positive cells were evaluated from P3 to P7, using Turnbull blue staining. The GFP-BMSC labeled by Ferucarbotran were transplanted into the ipsilateral striatum of the mice brain subjected to permanent focal ischemia at 7 days after the insult. The distribution and differentiation of GFP- and SPIO-positive cells in the brain were studied 3 months after transplantation, using immunohistochemistry and Turnbull blue staining. As the results, the proportions of the SPIO-positive cells gradually decreased from 93.6% at P3 to 6.5% at P7. Fluorescence immunohistochemistry revealed that the GFP-positive cells were widely distributed around infarct and partially expressed MAP2 and NeuN 3 months after transplantation. However, only a smaller number of SPIO-positive cells could be detected on Turnbull blue staining. The ratio of the SPIO- to GFP-positive cells was approximately 2.7%. The results strongly suggest that the BMSC repeat proliferation many times, migrate into the lesion, and partially express the neuronal phenotype in the host brain during 3 months after transplantation. The double labeling technique would be valuable to prove the proliferation of the transplanted cells in the host tissue because GFP gene and SPIO nanoparticles have different inheritance characteristics. (C) 2005 Elsevier B.V. All rights reserved.
  • S Yano, S Kuroda, JB Lee, H Shichinohe, T Seki, J Ikeda, G Nishimura, K Hida, M Tamura, Y Iwasaki
    JOURNAL OF NEUROTRAUMA 22 8 907 - 918 2005年08月 [査読有り][通常論文]
     
    Recent experimental studies have shown that bone marrow stromal cells (BMSC) differentiate into neural cells and reduce neurological deficits when transplanted into traumatized spinal cord. These findings have been derived primarily from histological analyses. We conducted a study directed chiefly at developing a non-invasive system for tracking BMSC transplanted into the spinal cord of living animals. In this study, we induced spinal cord injury (SCI) in rats with a pneumatic device. BMSC were harvested from transgenic mice expressing green fluorescence protein (BMSC-GFP), and were transplanted stereotactically into a control group of rats without SCI (n = 6) and a group with SCI (n = 3). At 2 and 4 weeks after transplantation, the dura mater was exposed and green fluorescence derived from the transplanted BMSC-GFP was observed. The distribution and differentiation of the transplanted cells were subsequently evaluated with immunohistochemistry. Green fluorescence could be detected around the transplantation site in three of six of the control rats. In all three rats subjected to SCI, green fluorescence was shown to spread from the site of BMSC-GFP injection toward the injury site, suggesting that the transplanted cells had migrated toward the lesion within the 4-week post-transplantation period. Histological evaluation suggested that the detected green fluorescence was emitted by cells that had distributed in the dorsal white matter, and demonstrated that some of the transplanted cells expressed neuronal or astrocytic markers. These results suggest the possibility of tracking BMSC transplanted into the spinal cord in living animals. Such noninvasive bioimaging techniques would be valuable for monitoring the fate of these transplanted cells and assessing the safety and efficacy of their transplantation.
  • H Shichinohe, S Kuroda, T Asano, S Ushikoshi, T Ishikawa, K Houkin, T Murashita, Y Iwasaki
    NEUROLOGICAL SURGERY 33 2 149 - 153 2005年02月 [査読有り][通常論文]
     
    Recently, there are increasing numbers of patients with occlusive carotid artery disease and coronary artery disease. Simultaneous or two-staged surgery for both lesions has been recommended for these patients to reduce the incidence of perioperative complications. However, therapeutic options for the patients with bilateral carotid artery stenosis and coronary artery disease have not been established. In this report, we describe two patients who a successfully underwent carotid endarterectomy (CEA) and carotid artery stenting (CAS) on each carotid artery in parallel with coronary artery bypass grafts (CABG). A 49-year-old male with severe stenosis of the bilateral internal carotid artery (ICA) and heart failure underwent CAS on the right side. Next day, he successfully under-went CABG and CEA on the left side at the same time. A 62-year-old male with severe stenosis of the bilateral ICA and coronary artery disease underwent CAS on the right side and CEA on the left side with an interval of 7 days. Subsequently, CABG was performed uneventfully. No perioperative complication occurred in either patient. The results suggest that combination therapy of CAS and CEA would be a valuable option for patients with complex carotid/coronary artery diseases.
  • 七戸 秀夫, 黒田 敏, 浅野 剛, 牛越 聡, 石川 達哉, 宝金 清博, 村下 十志文, 岩崎 喜信
    Neurological Surgery 33 2 149 - 153 (株)医学書院 2005年02月 
    症例1:49歳男.構音障害と右不全麻痺で発症し,両側内頸動脈に90%狭窄を認めると共に,慢性心不全,冠動脈三枝疾患の合併が判明した.冠動脈バイパス術(CABG)単独では両側大脳半球の灌流圧が著明に低下する危険が高いと考え,まず無症候性の右内頸動脈起始部の狭窄病変に対してstentを用いた血管形成術を施行した.翌日に左頸動脈内膜剥離術(CEA)および冠動脈バイパス術(CABG)を同時に行った.術後頸動脈の狭窄は改善し,両側性に脳血流量の増加を認め,5年経過し再発はない.症例2:62歳男.2年前,労作性狭心症でstentを併用した血管形成術を施行された.しかし,冠動脈狭窄が再発してCABGの適応となり,術前スクリーニングで両側の頸部内頸動脈に狭窄を指摘された.早期にCABGを実施する必要から,まず狭窄が比較的穏やかな右内頸動脈起始部の狭窄病変に対してSMART stentを留置し,1週間後に左CEAを施行した.その3週間後にCABGを施行し,周術期合併症はなかった
  • D Kashiwazaki, S Kuroda, S Terasaka, T Ishikawa, H Shichinohe, T Aoyama, S Ushikoshi, M Nunomura, Y Iwasaki
    NEUROLOGICAL SURGERY 33 1 29 - 34 2005年01月 [査読有り][通常論文]
     
    The authors report 9 patients who presented with loss of consciousness (syncope) due to occlusive carotid artery diseases. All patients were males, and their age ranged from 59 to 83 years. The attack was associated with dehydration or hypotension in 5 patients. MRI demonstrated fresh cerebral infarction in the watershed zone. Cerebral angiography revealed occlusion of the unilateral internal carotid artery (ICA) in 7 patients, severe stenosis of the bilateral ICA in one, and occlusion;of the unilateral ICA and severe stenosis of the contralateral ICA in one. Single photon emission tomography (SPECT) or positron emission tomography (PET) suggested reduced cerebral perfusion reserve because of inappropriate development of collateral circulation,in 4 out of 9 patients. These 4 patients underwent superficial temporal artery to middle cerebral artery anastomosis and/or carotid endarterectomy. Other 5 patients were medically treated. No further episode of syncope occurred in all 9 patients during follow-up periods. The results suggest that occlusive carotid artery diseases should be taken in considerations as a cause of syncope attack.
  • H Yasuda, H Shichinohe, S Kuroda, T Ishikawa, Y Iwasaki
    BRAIN RESEARCH 1032 1-2 176 - 182 2005年01月 [査読有り][通常論文]
     
    Previous studies have strongly suggested that heat shock protein 70 (HSP70) has protective effects in ischemia/reperfusion in tissues such as brain, heart, and liver. This study was performed to assess the efficacy of the HSP70 inducer geranylgeranylacetone (GGA) in experiments involving permanent middle cerebral artery (MCA) occlusion. Male Balb/c mice were subjected to permanent MCA occlusion by direct occlusion through small craniectomy. Vehicle or GGA (200 or 1000 mg/kg) was injected intraperitoneally 1 h prior to the onset of ischemia. Infarct volumes were evaluated at 24 h of ischemia by using 2,3,5-triphenyltetrazolium chloride (TTC) staining. The effect of GGA on the induction of HSP70 was studied at 3 h after ischemia with fluorescence immunocytochemistry. The percentage of infarct volume in the control mice (n=10) was 23.0 +/- 4.0% (mean +/- SD) of the contralateral hemisphere, while those in the treated groups were 22.6 +/- 7.3% (200 mg/kg group; n=5, P > 0.05) and 15.7 +/- 3.8% (1000 mg/kg group; n=5, P < 0.05). Pretreatments with 1000 mg/kg of GGA enhanced the ischemia-related induction of HSP in the neurons and astrocytes in the boundary zone of infarct. The results demonstrate that GGA significantly reduces infarct volume due to permanent MCA occlusion when given 1 h prior to the induction of ischemia. (C) 2004 Elsevier B.V. All rights reserved.
  • H Shichinohe, S Kuroda, H Yasuda, T Ishikawa, M Iwai, M Horiuchi, Y Iwasaki
    BRAIN RESEARCH 1029 2 200 - 206 2004年12月 [査読有り][通常論文]
     
    The present study was aimed to evaluate the effect of the free radical scavenger Edaravone on infarct volume due to permanent MCA occlusion in mice and, if so, to elucidate the mechanism of its neuroprotective effects. Male Balb/c mice were subjected to permanent middle cerebral artery occlusion and were treated with 3.0 mg/kg of Edaravone or vehicle 30 min before ischemia. Infarct volume was assessed by 2,3,5-triphenyltetrazolium chloride (TTC) method after 24 h. Furthermore, in situ detection of superoxide in the ipsilateral neocortex was carried out using the superoxide-sensitive dye dihydroethidium (DHE) staining technique. Pretreatment with 3.0 mg/kg of Edaravone ameliorated the tissue damage in the infarct rim and significantly reduced infarct volume to about 77% of the control (p<0.05). Semiquantitative measurement of red fluorescence emitted from DHE revealed that the superoxide increased in the ischemic core at 1 h after the onset of ischemia and extended towards the infarct rim at 3 and 6 h, and that pretreatment with 3.0 mg/kg of Edaravone significantly inhibited the increase of superoxide in the infarct rim at 3 and 6 h (p<0.01). Double staining with DHE and monoclonal antibody against NeuN showed that the majority of the nuclei positive for DHE were also positive for NeuN. These findings suggest that Edaravone salvages the boundary zone of infarct by scavenging reactive oxygen species especially in the neurons during permanent focal cerebral ischemia. (C) 2004 Elsevier B.V. All rights reserved.
  • JB Lee, S Kuroda, H Shichinohe, S Yano, H Kobayashi, K Hida, Y Iwasaki
    BRAIN RESEARCH PROTOCOLS 14 1 37 - 44 2004年11月 [査読有り][通常論文]
     
    In order to verify the biological aspects of 'autogeneic' bone marrow stromal cells (BMSC) transplantation for neurological disorders, we aimed our study towards the assessment of the survival, distribution, and differentiation of autologous BMSC in the central nervous system (CNS). We harvested rat BMSC from femur bones, and the nuclei were then fluorescently labeled by a 24-h co-culture with bis-benzimide. These BMSC were stereotactically injected into the striatum (n=6) or thoracic cord (n=8) of each animal. We evaluated the distribution and differentiation of 'autogeneic' BMSC in the brain and spinal cord after 4 weeks, using the immunohistochemistry technique. We found some injected cells in the ipsilateral striatum, hippocampus, neocortex, and bilateral corpus callosum, and approximately 20% and 15% of the engrafted cells expressed neuronal and astrocytic markers, respectively. Other injected cells were distributed in the dorsal funiculus and adjacent gray matter, and about 10% and 15% of these cells expressed neuronal and astrocytic markers, respectively. Although the precise mechanism of BMSC transdifferentiation still remains unclear, the present results show that 'autogeneic' BMSC could highly differentiate into their own CNS neural cells, suggesting that they are surrounded by favorable conditions. (C) 2004 Elsevier B.V. All rights reserved.
  • H Shichinohe, S Kuroda, JB Lee, G Nishimura, S Yano, T Seki, J Ikeda, M Tamura, Y Iwasaki
    BRAIN RESEARCH PROTOCOLS 13 3 166 - 175 2004年08月 [査読有り][通常論文]
     
    Recent experimental studies have indicated that bone marrow stromal cells (BMSC) improve neurological deficits when transplanted into the animal models of various neurological disorders, although precise mechanism still remains unclear. In this study, we developed a new in vivo fluorescence optical imaging protocol to sequentially track the transplanted into the brain of the living animals subjected to cerebral infarct. Mice BMSC were harvested from transgenic mice expressing green fluorescent protein (BMSC-GFP). They were stereotactically transplanted into the ipsilateral striatum of mice subjected to permanent middle cerebral artery occlusion after 7 days of ischemia (n = 12). During 12 weeks after transplantation, the skull was exposed and the green fluorescence emitted from the brain surface was sequentially observed, using in vivo fluorescence optical microscopy. As the results, regional green fluorescence was detected in the ipsilateral parietal region 4-12 weeks after transplantation in all animals and became more apparent over the time. The images obtained through the skull were very similar to those acquired by thinning or removing the skull. Immunohistochemistry evaluation revealed that the transplanted cells migrated towards the ischemic boundary zone and expressed the neuronal or astrocytic marker, supporting the findings on fluorescence optical images. Sequential visualization of the BMSC transplanted into the brain of living animals would be valuable for monitoring the migration, growth and differentiation of the transplanted cells to explore the fate and safety of stem cell transplantation for various neurological disorders. (C) 2004 Elsevier B.V. All rights reserved.
  • H Shichinohe, S Kuroda, T Abumiya, J Ikeda, T Kobayashi, T Yoshimoto, Y Iwasaki
    BRAIN RESEARCH 1001 1-2 51 - 59 2004年03月 [査読有り][通常論文]
     
    it has been reported that immunosuppressant FK506 inhibited ischemic neuronal injury in forebrain ischemia or transient focal cerebral ischemia, but the mechanisms of the neuroprotective effect have not been clarified. In permanent focal cerebral ischemia, we investigated whether FK506 caused remission of brain infarction, and how mechanism was concerned. Mate Balb/c mice were subjected to permanent middle cerebral artery (MCA) occlusion. They were treated with 1.0 or 3.0 mg/kg FK506 or vehicle 30 min before ischemia. Infarct volume was assessed by 2,3,5-triphenyltetrazolium chloride (TTC) method after 24 h. Cytochrome c release from mitochondria was evaluated by Western blotting and immunocytochemistry after ischemia. Simultaneously, the immunoreactivity of total and phosphorylated BAD was also studied using immunocytochemistry. We demonstrated that pretreatment with 3.0 mg/kg FK506 salvaged the tissue damage in the infarct rim and significantly reduced infarct volume to 75.5% (P < 0.05), and FK506 inhibited cytochrome e release on 6 h after ischemia for Western blot analysis (P < 0.05). Immunocytochemical study showed that permanent MCA occlusion increased the amount of cytochrome c and total BAD in the cytosol, but not phosphorylated BAD, in the ischemic core and the infarct rim as early as I h after ischemia, and FK506 inhibited the increases in the infarct rim. The results suggest that FK506 may, at least in part, ameliorate tissue damage due to permanent focal cerebral ischemia in the infarct rim through maintaining BAD turnover and inhibiting cytochrome c release from mitochondria. (C) 2004 Elsevier B.V. All rights reserved.
  • 黒田 敏, 七戸 秀夫, 小林 徹, 吉本 哲之, 鐙谷 武雄, 石川 達哉, 多田 光宏, 中井 章人, 宝金 清博, 岩崎 喜信
    Pharma Medica 22 2 130 - 130 (株)メディカルレビュー社 2004年02月
  • H Shichinohe, S Kuroda, T Ishikawa, Y Iwasaki
    NEUROLOGICAL SURGERY 32 1 75 - 78 2004年01月 [査読有り][通常論文]
     
    We present a rare case of jugular phlebectasia, which was able to be diagnosed by a color Doppler sonography. A 69-year-old female patient complained of a painless neck mass on the left side and consulted our hospital. MRI showed a round and regular mass with a diameter of 2.5cm. B-mode ultrasonography showed a soft, tubular mass in addition to the common carotid artery. In addition, color Doppler sonography revealed a very slow (about 6cm/sec) turbulent flow in the mass. Valsalva maneuver immediately stopped the slow flow in the mass and increased the diameter. These findings were immediately resolved after the termination of the Valsalva maneuver. The patient was diagnosed as having jugular phlebectasia, and was conservatively followed up. The specific findings of ultrasonography and color Doppler sonography were very useful to diagnose, non-invasively, jugular phlebectasia, a very rare disorder.
  • R Nanba, S Kuroda, M Takeda, H Shichinohe, N Nakayama, T Ishikawa, K Houkin, Y Iwasaki
    NEUROLOGICAL SURGERY 31 12 1291 - 1295 2003年12月 [査読有り][通常論文]
     
    Recent development of non-invasive diagnostic technology, such as magnetic resonance imaging (MRI) and angiography (MRA), is believed to have made possible on increase in the diagnoses of asymptomatic moyamoya disease. However, no criteria have been established for the management of such cases. The present study aimed to clarify the natural history of asymptomatic moyamoya disease retrospectively. Ten patients were included in this study. None of them had experienced any episode due to moyamoya disease and were only incidentally diagnosed as having moyamoya disease. There were 4 males and 6 females. Their ages ranged from 30 to 67 years, with the mean age of 46.2. Cerebral angiography showed there was the tendency of disease progression in elder patients. MRI detected cerebral infarction in 3 of 10 patients (30%). Hemodynamic ischemia, such as impaired reactivity to acetazolamide and/or elevated oxygen extraction fraction, was observed in 4 of 10 patients. Only one patient under-went surgical revascularization. Antiplatelet or anticonvulsant medication was administered in 5 of 10 patients. The mean follow-up period was 4.1 years, ranging from 0.5 to 13 years. During follow-up periods, the moyamoya lesion markedly progressed and caused cerebral infarction in one patient. However, neither ischemic nor hemorrhagic stroke occurred in the other 9 patients. Multi-center nation-wide study should be planned to clarify the natural course of asymptomatic moyamoya disease and establish the management guidelines for patients with asymptomatic moyamoya disease.
  • D Kashiwazaki, S Kuroda, S Ushikoshi, H Shichinohe, T Ishikawa, T Asano, T Shiga, N Tamaki, Y Iwasaki
    NEUROLOGICAL SURGERY 31 12 1315 - 1320 2003年12月 [査読有り][通常論文]
     
    There is increasing evidence that stenting is a useful strategy for internal carotid artery (ICA) stenosis in patients unfit for drastic surgery. However, it should be remembered that perioperative complications including seizure or intracerebral hemorrhage due to hyperperfusion are not so rare. The authors describe a case with severe ICA stenosis, who successfully underwent stenting as a result of intensive medical care for postoperative hyperperfusion. A 77-year-old man with a recent history of angina pectoris and transient ischemic attack was referred to our hospital. Cerebral angiography showed subtotal occlusion of the left ICA. SPECT/PET studies revealed a disturbed reactivity to acetazolamide and an increase in regional oxygen extraction fraction in the left hemisphere, suggesting a marked reduction in cerebral perfusion pressure. He successfully underwent carotid stenting. Intraoperative near-infrared monitoring showed an increase in the concentration of total and oxidized hemoglobin in the left frontal area after stenting. A SPECT study just after stenting also demonstrated hyperperfusion in the left middle cerebral artery territory. His blood pressure was carefully controlled to avoid "hyperperfusion syndrome" including headache, seizure and intracerebral hemorrhage. Follow-up SPECT/PET studies showed a normalization of hemodynamic and metabolic parameters. SPECT/PET studies are quite valuable to predict and prevent hyperperfusion syndrome after carotid stenting, and result in good clinical outcome.
  • 難波 理奈, 黒田 敏, 竹田 誠, 七戸 秀夫, 中山 若樹, 石川 達哉, 宝金 清博, 岩崎 喜信
    Neurological Surgery 31 12 1291 - 1295 (株)医学書院 2003年12月 
    過去10年間に著者らが経験した成人無症候性もやもや病10例(男性4例,女性6例・平均年46.2歳)の臨床像,予後について検討した.脳MRIでは脳梗塞を3例に認め,梗塞部位はMCA-ACAあるいはMCA-PCA境界域,尾状核であった.DSAでは,全例病変は両側に存在し,鈴木の病期分類は30歳代で1〜3期が多く,50歳以降では4〜6期の例が多い傾向にあった.SPECTやPETによる脳循環代謝パラメータでは,内頸動脈領域の脳虚血を4例に認め,うち2例は脳梗塞を有した.平均4.1年間の経過観察中,1例でもやもや病の病期が進行し,脳梗塞を発症したが,その他の9例は経過観察期間中に脳虚血発作を呈した例はなく,うち1例は診断2年目に妊娠し,帝王切開にて無事出産した
  • JB Lee, S Kuroda, H Shichinohe, J Ikeda, T Seki, K Hida, M Tada, K Sawada, Y Iwasaki
    NEUROPATHOLOGY 23 3 169 - 180 2003年09月 [査読有り][通常論文]
     
    There is increasing evidence that bone marrow stromal cells (BMSC) have the potential to migrate into the injured neural tissue and to differentiate into the CNS cells, indicating the possibility of autograft transplantation therapy. The present study was aimed to clarify whether the mouse BMSC can migrate into the lesion and differentiate into the CNS cells when transplanted into the mice subjected to focal cerebral infarct or spinal cord injury. The BMSC were harvested from mice and characterized by flow cytometry. Then, the BMSC were labeled by bis-benzimide, a nuclear fluorescence dye, over 24 h, and were stereotactically transplanted into the brain or spinal cord of the mice. The cultured BMSC expressed low levels of CD45 and high levels of CD90 and Sca-1 on flow cytometry. A large number of grafted cells survived in the normal brain 4 weeks after transplantation, many of which were located close to the transplanted sites. They expressed the neuronal marker including NeuN, MAP2, and doublecortin on fluorescent immunohistochemistry. However, when the BMSC were transplanted into the ipsilateral striatum of the mice subjected to middle cerebral artery occlusion, many of the grafted cells migrated into the corpus callosum and injured cortex, and also expressed the neuronal markers 4 weeks after transplantation. In particular, NeuN was very useful to validate the differentiation of the grafted cells, because the marker was expressed in the nuclei and was overlapped with bis-benzimide. Similar results were obtained in the mice subjected to spinal cord injury. However, many of the transplanted BMSC expressed GFAP, an astrocytic protein, in injured spinal cord. The present results indicate that the mouse BMSC can migrate into the CNS lesion and differentiate into the neurons or astrocytes, and that bisbenzimide is a simple and useful marker to label the donor cells and to evaluate their migration and differentiation in the host neural tissues over a long period.
  • J Kitamura, S Kuroda, S Ushikoshi, K Furukawa, T Asano, H Shichinohe, K Houkin, Y Iwasaki, H Saitoh, K Mitsumori
    NEUROLOGICAL SURGERY 30 10 1097 - 1102 2002年10月 [査読有り][通常論文]
     
    We report three cases of radiation-induced carotid arterial stenosis that underwent successful angioplasty with stenting. The patients had received radiation therapy for tongue or laryngeal cancers and developed minor completed strokes 6 to 14 years after irradiation. All patients had multiple and bilateral stenosis, measuring more than 50%, of the carotid arteries. The stenosis was located in the internal, external, and common carotid arteries. We performed percutaneous transluminal angioplasty with stenting. All interventions were successful and carotid stenosis decreased to less than 28%. No permanent complications occurred. During follow-up periods of Lip to 26 months, all of these cases were free from ischemic symptoms. Neither carotid angiography nor ultrasound sonography showed evidence of restenosis. The present results suggest the usefulness of angioplasty with stenting for radiation-induced carotid arterial steriosis.
  • 北村 淳, 黒田 敏, 牛越 聡, 古川 浩司, 浅野 剛, 七戸 秀夫, 宝金 清博, 岩崎 喜信, 斉藤 久壽, 三森 研自
    Neurological Surgery 30 10 1097 - 1102 (株)医学書院 2002年10月 
    舌癌或いは喉頭癌に対して放射線治療の既往があり,頸部内頸動脈狭窄症を認めた3例を対象とし,ステントを併用した血管形成術を施行した.ステントを併用した経皮的血管形成術(PTA)により,全ての病変において十分な狭窄の改善が得られた.狭窄率は,いずれも28%以下と著明に改善した.周術期に脳虚血症状は出現しなかった.経過観察期間中に,DSA又は頸動脈エコーにおいて明らかな再狭窄は認められず,新たな脳梗塞の再発も観察されなかった.合併症は1例で見られ,PTAを行った直後に痙攣が出現したが,その後,消失した
  • もやもや病の脳循環代謝 SPECT/PETによる検討
    七戸 秀夫, 黒田 敏, 宝金 清博, 岩崎 喜信, 志賀 哲, 加藤 千恵次, 玉木 長良
    脳循環代謝 14 1 81 - 81 (一社)日本脳循環代謝学会 2002年03月
  • Y Ohta, H Shichinohe, K Nagashima
    NEUROLOGIA MEDICO-CHIRURGICA 42 1 40 - 43 2002年01月 [査読有り][通常論文]
     
    A 69-year-old woman with a 14-year history of polycythemia vera suffered progressive paraparesis due to epidural involvement of hematopoietic tissue. Magnetic resonance (MR) imaging demonstrated extensive epidural masses. Decompressive surgery and radiotherapy were performed and she made an almost complete clinical recovery. Serial MR imaging showed no regrowth of the other epidural masses. Extramedullary hematopoiesis occurs in patients with various hematologic disorders involving a chronic increase in the production of red blood cells, and is often associated with thalassemia, but is less common with polycythemia vera. The most frequent sites are the spleen, liver, and kidney. Extramedullary hematopoietic tissue occurring within the spinal canal and causing cord compression is very rare. Total surgical excision is not usually feasible because of the diffuse nature of extramedullary hematopoietic tissue and the possibility of recurrence, but acute neurological deterioration does require emergency surgery. Extramedullary hematopoiesis is radiosensitive and displays a rapid response to low dosages, so radiation therapy is recommended for residual tumors. Considering the possibility of central nervous system extramedullary hematopoiesis in patients with polycythemia vera, an early diagnosis is necessary for a favorable prognosis.
  • 七戸 秀夫, 黒田 敏, 宝金 清博, 牛越 聡, 岩崎 喜信, 阿部 弘
    Neurological Surgery 29 9 871 - 876 (株)医学書院 2001年09月 
    視野障害を合併する内頸動脈高度狭窄症に対して,内頸動脈stent留置術を施行し,術前に低下していた後大脳動脈領域の脳血流量が増加し,症状が改善した症例を2例経験した.2例に共通する病態として,脳血管撮影では内頸動脈に高度狭窄が存在し,同側の後交通動脈が太く,これによって内頸動脈領域への側副血行路がよく発達していた.この為,SPECTでは反対側の後大脳動脈の脳血流量やacetazolamide反応性が低下し,その後頭葉に起因する視野狭窄を認めた.症例1(64歳男)では後頭葉に小さな脳梗塞もみられた.頸動脈狭窄症に対してstentを併用した経皮的血管形成術を実施したところ,後頭葉の脳循環が改善すると共に視野障害が軽快した.特に,症状の改善は脳梗塞を有さなかった症例2(64歳男)において顕著であった.いずれの症例においても椎骨動脈の起始部やその遠位部に閉塞性病変は認められなかった
  • H Shichinohe, S Kuroda, K Houkin, S Ushikoshi, Y Iwasaki, H Abe
    NEUROLOGICAL SURGERY 29 9 871 - 876 2001年09月 [査読有り][通常論文]
     
    Although previous reports have suggested "steal VBI" due to occlusive carotid artery diseases, there have been no reports that clearly define "steal VBI" from the viewpoint of cerebral hemodynamics. The authors presented two cases with "steal VBI" due to severe stenosis of the internal carotid artery. Both patients had well-developed collateral circulation through the ipsilateral posterior communicating artery. Although no occlusive lesion was found in the vertebrobasilar system, blood flow studies revealed impaired hemodynamics in the contralateral occipital lobe, which fact correlated with their neurological deficit, visual field disturbance. Carotid stenting markedly corrected the stenotic lesions, leading to neurological improvement. Follow-up blood studies showed normalization of hemodynamics in the contralateral occipital lobe. The findings strongly suggest that carotid surgery or stenting can improve cerebral hemodynamics in the carotid systems, resolving "steal VBI" due to developed collaterals from the posterior to the anterior circulation.
  • 七戸 秀夫, 黒田 敏, 宝金 清博, 岩崎 慶信, 布村 充
    脳卒中 23 1 136 - 136 (一社)日本脳卒中学会 2001年03月
  • 七戸 秀夫, 黒田 敏, 宝金 清博, 阿部 弘, 菊池 陽一, 牛越 聡
    脳卒中 22 1 282 - 282 (一社)日本脳卒中学会 2000年04月
  • 七戸 秀夫, 黒田 敏, 石川 達哉, 宝金 清博, 阿部 弘, 松居 喜郎, 牛越 聡, 菊池 陽一
    新潟医学会雑誌 113 11〜12 575 - 575 新潟医学会 1999年12月
  • 宝金 清博, 七戸 秀夫, 黒田 敏, 松居 喜朗, 牛越 聡
    治療学 33 5 582 - 584 ライフサイエンス出版(株) 1999年05月
  • 脳動脈瘤破裂による急性期くも膜下出血のMRI,MRA診断
    青樹 毅, 金子 貞男, 七戸 秀夫, 宝金 清博, 阿部 弘, 三森 研自
    日本脳神経外科学会総会抄録集 56回 28 - 28 (一社)日本脳神経外科学会 1997年10月

その他活動・業績

共同研究・競争的資金等の研究課題

  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2020年04月 -2023年03月 
    代表者 : 川堀 真人, 七戸 秀夫
     
    昨年度製法が確立したラット脳出血モデル（SDラットおよび免疫抑制ラット）に対して出血が吸収された2週後に出血腔内へのセルザイク投与（ラット由来およびヒト由来）を行い、その後1か月間の運動機能改善を評価した。またIn-vitroにおいてセルザイクとBMSCの栄養因子の分泌について評価した。In-vivoでセルザイク投与群の方がBMSC/生理食塩水より神経機能改善効果が有意に高く、脳萎縮率もCellSaic群で低い傾向であった。また、セルザイク投与群での血腫腔周囲の脳由来神経栄養因子（BDNF）量は生理食塩水群よりも有意に多く、BDNF抑制セルザイクは通常のセルザイクよりも神経機能改善の程度が小さかった。OGD環境では、CellSaicの方がBMSCより栄養因子分泌能および細胞増殖能が有意に高かった。
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2016年04月 -2019年03月 
    代表者 : 鐙谷 武雄, 小松 晃之, 中山 若樹, 数又 研, 七戸 秀夫, 寳金 清博, 長内 俊也
     
    2時間脳虚血後に10℃に冷却した人工酸素運搬体HemoActを再開通動脈から投与した。この人工酸素運搬体投与＋冷却灌流のcombination therapy群を、10℃生理食塩水の同量投与、37℃HemoActの同量投与の二つmonotherapy群と、未治療コントロール群と比較して検討した。その結果、combination therapyの方が梗塞体積を縮小し、脳血流量低下を抑制する傾向にあった。この治療効果は、少なくとも治療7日後まで継続した。また、therapeutic time windowの延長効果の検討では、虚血時間が5時間までは治療効果があることが分かった。
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2015年04月 -2018年03月 
    代表者 : 関 俊隆, 笹森 徹, 岩崎 素之, 七戸 秀夫, 寳金 清博
     
    脊髄癒着性くも膜炎は，くも膜・軟膜に慢性炎症性変化をきたし，慢性進行性に脊髄を障害する．確立された治療法はなく，きわめて治療困難である．難治性疼痛を認めることは稀ではなく，患者の日常生活を著しく損なう．本研究は脊髄損傷後の脊髄癒着性くも膜炎に起因する難治性疼痛の治療法の確立を目的とした．本実験の前に安定した脊髄損傷動物実験モデルを作製することとした．既存の動物脊髄損傷用の脳動脈瘤クリップは高価で継続的な実験が困難であったため，本邦の企業に依頼し動物脊髄損傷用の脳動脈瘤クリップを作製した．既存のクリップと比較して統計学的にも有意差のない脊髄損傷モデルを作製することできた．
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2015年04月 -2018年03月 
    代表者 : 数又 研, 中山 若樹, 鐙谷 武雄, 七戸 秀夫, 寳金 清博
     
    iPS細胞より血管内皮細胞を分化誘導した。マイクロアレイを行った結果、もやもや病血管内皮細胞において、tight junction関連遺伝子であるCLDN1, Focal adhesions and hemidesmosomes関連遺伝子であるDST, NEXN, ITGB3が低下していた。RT-PCRでは、ITGB3はもやもや病血管内皮細胞において発現が低下していたが、DST, NEXNは有意な差はなく、CLDN1は明らかなバンドが確認できなかった。血管透過性アッセイにおいてY-27632を添加した際に、健常群では透過性が低下したのに対し、もやもや病においては亢進した。
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2015年04月 -2018年03月 
    代表者 : 穂刈 正昭, 七戸 秀夫, 中山 若樹, 寳金 清博
     
    くも膜下出血急性期の炎症反応を制御し、神経損傷を軽減することが本研究の目的である。重症くも膜下出血モデルを作成し、それに対して低体温療法・ミノサイクリン投与を行うことによって、急性期炎症反応の制御および予後改善が可能であるか検討する。本研究を通じてくも膜下出血後の炎症反応に早期低体温療法は運動機能改善を伴う炎症反応の低下を認めたが、minocyclineは炎症反応の低下は認めるものの運動機能回復は認められないという結果であった。
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2014年04月 -2018年03月 
    代表者 : 久下 裕司, 東川 桂, 川井 恵一, 浅田 祐士郎, 鐙谷 武雄, 七戸 秀夫, 山下 篤, 玉木 長良, 西嶋 剣一, 志水 陽一
     
    本研究では、インビトロ、インビボ実験により以下の成果を得た。 ①インテグリンαvβ3イメージング剤18F-RGD-K5の自動合成法を確立した。②低酸素イメージング剤18F-FMISOが、18F-FDGとは異なり、低酸下のM2マクロファージに集積することを見出した。③18F-FMISOの母体構造である2-nitroimidazole系化合物が動脈硬化病変に集積することを確認した。 これらの成果は、臨床診断に役立つ動脈硬化イメージング法の構築に資するものである。
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2015年04月 -2017年03月 
    代表者 : 寳金 清博, 中山 若樹, 数又 研, 鐙谷 武雄, 七戸 秀夫
     
    もやもや病の病態を解析するため、もやもや病特異的iPS細胞から平滑筋細胞を分化誘導した。まず、iPS細胞から神経堤細胞を分化誘導し、それらの細胞は99%以上でp75陽性であることを確認した。それらの細胞に対して平滑筋細胞へと分化誘導を行ったが、分化誘導の途中で細胞の老化、増殖能力の低下が認められた。最終的に残った細胞に対して免疫染色を行ったところ、ほぼ全ての細胞でα-SMA、SM22陽性であり、平滑筋細胞へと分化していることが示された。しかし、細胞の増殖能力低下のため、解析に必要な十分な細胞数を得ることが困難であった。今後、細胞の老化を防ぐために分化誘導方法を改良する必要があると考えられる。
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2013年04月 -2016年03月 
    代表者 : 吉本 哲之, 中山 若樹, 数又 研, 七戸 秀夫, 寳金 清博, 佐々木 秀直, 伊東 雅基
     
    成人発症もやもや病の親から、小児もやもや病の子が生まれた場合（臨床的表現促進現象）、子はより顕著なもやもや病の遺伝的素因を保有している可能性がある。もやもや病親子例を対象として、全ゲノム網羅的マイクロアレイ遺伝子コピー数多型(CNV)解析を実施した。もやもや病親子15組中、表現促進を認めた10組では、16番染色体短腕上(16p13.3)の410kbpの領域にCNVを認める頻度が高く、遺伝子コピー数が、親世代より子世代で増幅している傾向を認めた。遺伝子コピー数の世代を超えた増幅が臨床的表現促進現象の背景となりうることが示唆され、次世代シーケンス解析による、より詳細な検証が必要である。
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2013年04月 -2015年03月 
    代表者 : 寳金 清博, 中山 若樹, 数又 研, 鐙谷 武雄, 七戸 秀夫
     
    本研究では、ウシBMECの過継代による老化モデル実験を行った。過継代P15～20 BMECにおいて巨細胞が出現し、β-galactosidase活性、活性酸素産生が増大していた。次いで、P5 BMECとP15 BMECで細胞の生死、アポトーシス、SIRT1発現を比較した。未刺激では差がなかったが、OGD/再酸素化の負荷刺激をすると、P15 BMECで細胞死、アポトーシスが増加した。一方、SIRT1発現は、P5 BMECでのみ負荷刺激後に増加した。この結果はSIRT1が細胞保護的に働いている可能性を示し、老化細胞でSIRT1の誘導がかかりにくいことが細胞の脆弱性に関連していることを想起させた。
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2012年04月 -2015年03月 
    代表者 : 寳金 清博, 黒田 敏, 七戸 秀夫
     
    1) 患者血中EPCのFACS解析：患者33名、健康対照14名から採血を行った。術前では小児患者、成人患者ともにEPCの有意な細胞数低下が示された。また、術後は有意なCD34+/CD133+細胞の低下がみられた。これら所見は、病変部でのEPC消費亢進を示唆していると考えられた。 2) もやもや病患者由来iPS細胞の樹立：患者3名、正常対照3名からiPS細胞を作成した。血管内皮細胞への分化誘導を行い検討したところ、患者細胞では管腔形成能の低下がみられた。マイクロアレイとプロテオミクス解析では、患者で発現が低下している遺伝子/タンパク質に血管新生に関与するものが見出され、病因との関連が示唆された。
  • 文部科学省：科学研究費補助金(基盤研究(B))
    研究期間 : 2011年 -2012年 
    代表者 : 七戸 秀夫
     
    ボランティア由来のBMSC(bone marrow stromal cells)を使用した。血小板濃厚液からPL(platelet lysate)を作成し、基本培地に加えてBMSCを培養した。PLで培養したBMSCは培養速度、栄養因子産生においてFCS(仔ウシ血清)で培養したものとほぼ同等であった。またPLで培養したBMSCは、ラット脳梗塞モデルへの移植後の運動機能回復においてもFCSで培養したものと同等以上であった。また小動物用MRIや18F-FDG PETで、ラット脳梗塞モデルへの移植後の細胞追跡や脳機能回復の画像化を試みた。これらの結果は、次世代の臨床試験プロトコールに取り入れられる。
  • 文部科学省：科学研究費補助金(基盤研究(B))
    研究期間 : 2009年 -2011年 
    代表者 : 黒田 敏, 飛騨 一利, 小林 浩之, 七戸 秀夫
     
    ラット大脳脳挫傷モデルにおいてフィブリンがBMSC移植の際の有用なバイオマテリアルであることを証明した(J Neurosurg誌2010)。新規ポリマーを用いることでマウス脳梗塞モデルへのBMSC移植がより効率的に実施できることを確認した(Neurosurgery誌2010)。骨髄から得られる単核球分画にはBMSCのような脳梗塞に対する再生能力が乏しいことを発見した(Neuropathology誌2010)。慢性脳虚血モデルにおける認知機能の低下をBMSCの脳内移植によって抑制できることを世界で初めて発見した(Dement Geriatr Cogn Disord誌2010)。長波長蛍光物質でBMSCを標識してラット脳梗塞モデルに移植することで頭皮上から移植細胞を可視化した(Neurosurgery誌2011)。ラット脳挫傷モデルにBMSCを頚動脈から移植することで神経症状の改善を認めた(Neurosurgery誌投稿中)。老齢ラットのBMSCをG-CSFとともに培養することでその増殖や移植による治療効果を増強することができることを証明した(Cytokine誌投稿中)。ヒトBMSCを動物蛋白を用いない方法で培養してラット脳梗塞モデルに移植し、その治療効果を証明した(Neurosurgery誌2011)。同様の効果を小動物用MRIにて生体内でも確認した(Cell Transpla...
  • 文部科学省：科学研究費補助金(基盤研究(B))
    研究期間 : 2008年 -2010年 
    代表者 : 七戸 秀夫, 矢野 俊介, 黒田 敏, 岩崎 喜信, 飛騨 一利, 小林 浩之
     
    ヒト骨髄間質細胞(BMSC)を利用した中枢神経再生を臨床応用させるため、その効率的な手法の確立と安全性の追求を目指した研究を行い、以下の成果を得た。(1)バイオマトリックスを用いたBMSC培養に成功した。(2)G-CSF(顆粒球コロニー刺激因子)を培養液中に添加すると、BMSCが活性化された。またヒトPL(血小板溶解物)を添加し、動物由来成分を排除した培養を行いその有用性を示した。(3)フィブリンマトリックスやメビオールジェルRはスキャフォールドとして有用であり、病変部へBMSCを効果的にデリバリーすることを示した。(4)移植後のヒトBMSCの挙動を、光イメージング技術やMRIにて非侵襲的、経時的にモニタリングできた。
  • 文部科学省：科学研究費補助金(基盤研究(C))
    研究期間 : 2008年 -2010年 
    代表者 : 矢野 俊介, 飛騨 一利, 黒田 敏, 七戸 秀夫, 岩崎 喜信
     
    この3年間に表記の研究課題に関して数多くの知見を得ることができた。すなわち、自己骨髄から分離した骨髄間質細胞(bone marrow stromal cells;BMSC)の生物学的特性を解明するとともに、MRI、光イメージングなどを多用することで、臨床応用を視野に入れた安全かつ効率的な細胞の培養法、中枢神経への投与法、治療効果判定のための画像診断法を確立するための基礎データを得ることができた。
  • 文部科学省：科学研究費補助金(基盤研究(B))
    研究期間 : 2005年 -2007年 
    代表者 : 飛騨 一利, 七戸 秀夫, 黒田 敏, 岩崎 喜信, 矢野 俊介, 小林 浩之, 中山 若樹
     
    蛍光標識した骨髄間質細胞(BMSC)をラット脊髄損傷モデルに移植して、生体において移植されたBMSCを可視化する技術を開発した(J Neurotrauma誌2005)。GFP遺伝子と鉄製剤で標識したBMSCをマウス脳梗塞モデルに移植して、BMSCが脳内で増殖を繰り返していることを証明した(Brain Res誌2005)。ラット脊髄損傷モデル、マウス脳梗塞モデルに移植されたBMSCが神経細胞に分化することで、病変周囲のGABA受容体機能を改善させることをオートラジオグラフィを用いて証明した(J Nucl Med誌2006,J Neurotrauma誌2006)。BMSCの遺伝子プロファイリングを解析してBMSCが神経細胞に分化する能力を有していることを証明した(Brain Res誌2006)。BMSCが有するCXCR4受容体が、脳梗塞周囲で産生されるSDF-1を感知して、脳梗塞周囲に遊走することをノックアウトマウスを用いて証明した(Brain Res誌2007)。BMSCは神経細胞と共存することで神経細胞に分化する能力を有していること、BMSCはさまざまな神経栄養因子を産生することで興奮性アミノ酸に暴露された神経細胞を保護することを、新たに開発した共培養システムを用いて証明した(J Neurosci Res誌2008)。ラット脊髄スライスに移植したBMSCが脊髄の基質を溶解させ...
  • 文部科学省：科学研究費補助金(基盤研究(C))
    研究期間 : 2005年 -2006年 
    代表者 : 七戸 秀夫, 小林 浩之, 小林 浩之, 黒田 敏, 飛騨 一利, 矢野 俊介, 岩崎 喜信, 菊地 誠志
     
    マウス脳梗塞モデル、および、ラット脊髄損傷モデルに骨髄間質細胞(bone marrow stromal cell ; BMSC)を定位的に移植すると、BMSCが病変周囲に遊走して神経細胞特有の表現型を獲得するのみならず、神経細胞にのみ存在する受容体の機能を獲得して、脳梗塞や脊髄損傷周囲における受容体機能を改善させることを明らかとした。これらの結果は、それぞれJ Nucl Med誌(2006)、J Neurotrauma誌(2006)に掲載された。BMSCが神経細胞に分化する際の分子生物学的背景を明らかにする目的で、培養BMSCのマイクロアレイ解析を実施した。その結果、BMSCは神経細胞への分化誘導によって、その遺伝子表現型を変化させることが判明した。この結果はBrain Res誌(2006)に掲載された。BMSCが病変周囲に遊走するためには病変周囲で産生されるSDF-1、および、BMSCの膜表面に存在する3DF-1に特異的受容体であるCXCR4が必須の役割を果たしていることを、CXCR4ノックアウトマウスを用いた実験を実施することで明らかとした。現在、J Cereb Blood Flow Metab誌に投稿中である。BMSCと神経細胞の共培養実験系を確立した。この実験系を活用することにより、BMSCの一部が神経細胞そのものへ分化すること、一部は神経細胞と細胞融合することを初め...
  • 文部科学省：科学研究費補助金(萌芽研究)
    研究期間 : 2005年 -2006年 
    代表者 : 岩崎 喜信, 七戸 秀夫, 鐙谷 武雄, 石川 達哉, 黒田 敏, 中山 若樹
     
    マウス脳梗塞モデル、および、ラット脊髄損傷モデルに骨髄間質細胞(bone marrow stromal cell; BMSC)を定位的に移植すると、BMSCが病変周囲に遊走して神経細胞特有の表現型を獲得するのみならず、神経細胞にのみ存在する受容体の機能を獲得して、脳梗塞や脊髄損傷周囲における受容体機能を改善させることを明らかとした。これらの結果は、それぞれJ Nucl Med誌(2006)、J Neurotrauma誌(2006)に掲載された。BMSCが神経細胞に分化する際の分子生物学的背景を明らかにする目的で、培養BMSCのマイクロアレイ解析を実施した。その結果、BMSCは神経細胞への分化誘導によって、その遺伝子表現型を変化させることが判明した。この結果はBrain Res誌(2006)に掲載された。BMSCが病変周囲に遊走するためには病変周囲で産生されるSDF-1、および、BMSCの膜表面に存在するSDF-1に特異的受容体であるCXCR4が必須の役割を果たしていることを、CXCR4ノックアウトマウスを用いた実験を実施することで明らかとした。現在、J Cereb Blood Flow Metab誌に投稿中である。BMSCと神経細胞の共培養実験系を確立した。この実験系を活用することにより、BMSCの一部が神経細胞そのものへ分化すること、一部は神経細胞と細胞融合することを初めて...
  • 文部科学省：科学研究費補助金(基盤研究(B))
    研究期間 : 2004年 -2006年 
    代表者 : 石井 伸明, 岩崎 喜信, 岩崎 喜信, 小林 浩之, 黒田 敏, 七戸 秀夫, 矢野 俊介, 吉野 雅美
     
    ラットおよびマウス骨髄からBMSCを分離培養する実験系を確立した。FACSを用いた解析ではCD34(-)、CD45(+/-)、CD90(+)、Sca1(+)であり、間葉系細胞であることがわかった。Brain Res Protoc誌(2004)に掲載。移植されたBMSCの挙動に関しマウス・ラット脳梗塞・脊髄損傷モデルで検討した。移植細胞は活発な細胞分裂を繰り返し生着することが明らかとなった。Brain Res誌(2005)に掲載。マウスから分離・培養したBMSCの培養液にDMSO, retinoic acid, bFGFを添加して分化誘導を試みた。その結果、BMSCは形態学的には神経細胞に類似したものに変化し、nestinやTuj-1など未分化な神経細胞に特異的なマーカーを発現することが免疫染色により確認された。また、DNAマイクロアレイにより、分化誘導前後におけるBMSCの遺伝子発現プロファイルを検討したところ、BMSCは外的刺激によりその遺伝子発現を変化させ得ること、様々な成長因子を産生することで損傷された中枢神経を保護する役割を有することが示唆された。Brain Res誌(2006)に掲載。細胞移植を促進するためのバイオマテリアルに関して検討したところフィブリンが有用な素材となり得ることが明らかとなった。(投稿準備中)BMSCと神経細胞の融合能および分化能の評価をin v...
  • 文部科学省：科学研究費補助金(基盤研究(B))
    研究期間 : 2003年 -2005年 
    代表者 : 岩崎 喜信, 関 俊隆, 池田 潤, 小林 浩之, 黒田 敏, 七戸 秀夫, 飛騨 一利, 田村 守, 矢野 俊介
     
    マウスおよびラット大腿骨より骨髄間葉系細胞(BMSC)を分離し、マウス脳梗塞あるいは脊髄損傷モデルに移植する実験系を確立し、Neuropathology誌(2003)、Brain Res Protoc誌(2004)に掲載された。GFPトランスジェニック・マウスからBMSCを採取してマウス脳梗塞あるいはラット脊髄損傷モデルに移植した。移植した細胞が脳や脊髄の中を病変部に向かって遊走する様子を動物が生きたままの状態で観察する実験系を確立した。Brain Res Protoc誌(2004)、J Neurotrauma誌(2005)に掲載された。移植されたBMSCが神経細胞に特異的に存在する受容体を保護するかどうかを、マウス脳梗塞モデル、ラット脊髄損傷モデルを用いてautoradiographyにより解析した。細胞移植が病変周囲における神経受容体を保持する役割を有していることが判明した。J Nucl Med誌(2006)に掲載された。さらにJ Neurotrauma誌に投稿中である。移植されたBMSCが神経回路網を再構築する様子を生体内で観察する目的で、MRIによるマウス脳梗塞モデル、ラット脊髄損傷モデルの画像化システムを確立した。移植されたBMSCが中枢神経内部で活発な細胞分裂を繰り返しながら生着することを明らかとした。Brain Res誌(2005)に掲載された。仔ラット脊髄スラ...
  • 文部科学省：科学研究費補助金(基盤研究(B))
    研究期間 : 2002年 -2005年 
    代表者 : 黒田 敏, 飛騨 一利, 田村 守, 澤田 賢一, 池田 潤, 多田 光宏, 矢野 俊介, 関 俊隆, 七戸 秀夫, 岩崎 喜信, 飛騨 一利, 田村 守
     
    マウスおよびラット大腿骨より骨髄間葉系細胞(BMSC)を分離し、マウス脳梗塞あるいは脊髄損傷モデルに移植する実験系を確立し、Neuropathology誌(2003)、Brain Res Protoc誌(2004)に掲載された。GFPトランスジェニック・マウスからBMSCを採取してマウス脳梗塞あるいはラット脊髄損傷モデルに移植した。移植した細胞が脳や脊髄の中を病変部に向かって遊走する様子を動物が生きたままの状態で観察する実験系を確立した。Brain Res Protoc誌で2004)、J Neurotrauma誌(2005)に掲載された。移植されたBMSCが神経細胞に特異的に存在する受容体を保護するかどうかを、マウス脳梗塞モデル、ラット脊髄損傷モデルを用いてautoradiographyにより解析した。細胞移植が病変周囲における神経受容体を保持する役割を有していることが判明した。J Nucl Med誌(2006)に掲載された。さらにJ Neurotrauma誌に投稿中である。移植されたBMSCが神経回路網を再構築する様子を生体内で観察する目的で、MRIによるマウス脳梗塞モデル、ラット脊髄損傷モデルの画像化システムを確立した。移植されたBMSCが中枢神経内部で活発な細胞分裂を繰り返しながら生着することを明らかとした。Brain Res誌(2005)に掲載された。仔ラット脊髄スラ...

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  • 医倫理学序論
    開講年度 : 2021年
    課程区分 : 修士課程
    開講学部 : 医学院
    キーワード : 生命倫理、ヘルシンキ宣言、研究倫理、研究不正、疫学研究、臨床研究、医療情報、遺伝子解析、遺伝子治療 bioethics, Declaration of Helsinki, ethics of science and technology, research misconduct, epidemiological studies, clinical trials, medical informatics, human genome and genetic sequencing research, genome editing.
  • 医倫理学
    開講年度 : 2021年
    課程区分 : 博士後期課程
    開講学部 : 医学院
    キーワード : 生命倫理、ヘルシンキ宣言、研究倫理、研究不正、疫学研究、臨床研究、医療情報、遺伝子解析、遺伝子治療 bioethics, Declaration of Helsinki, ethics of science and technology, research misconduct, epidemiological studies, clinical trials, medical informatics, human genome and genetic sequencing research, genome editing.
  • トランスレーショナルリサーチ概論
    開講年度 : 2021年
    課程区分 : 学士課程
    開講学部 : 医学部
    キーワード : トランスレーショナルリサーチ 臨床研究

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