研究者詳細 - 緒方　藍歌

Bone marrow-derived mesenchymal stromal cells regress aortic aneurysm via the NF-kB, Smad3 and Akt signaling pathways. 査読有り

＜Editors' Choice＞ Effects of exosomes derived from the induced pluripotent stem cells on skin wound healing. 招待有り査読有り

Kobayashi H, Ebisawa K, Kambe M, Kasai T, Suga H, Nakamura K, Narita Y, Ogata A, Kamei Y

Nagoya journal of medical science 80 巻 ( 2 ) 頁： 141 - 153 2018年5月

Yamawaki-Ogata A, Oshima H, Usui A, Narita Y

Cytotherapy 19 巻 ( 10 ) 頁： 1167 - 1175 2017年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Cytotherapy

Background aims We have confirmed that aortic aneurysm (AA) can be regressed by the administration of bone marrow–derived mesenchymal stem cells (BM-MSCs). We investigated the kinetics of signaling pathways in AA following treatment with BM-MSCs. Methods Angiotensin II-infused apolipoprotein E–deficient mice were treated by intravenous injection of 1 × 106 BM-MSCs in 0.2 mL saline (BM-MSCs group, n = 5) or 0.2 mL saline (saline group, n = 5). Mice were sacrificed 2 weeks after injection and subjected to measurements of the incidence of AA and levels of phosphorylated proteins. Levels of proteins in conditioned media of BM-MSCs were also measured. Results The incidence of AA in the BM-MSCs group was reduced (BM-MSC 40% versus saline 100%, P < 0.05). Levels of pNF-kB and pSTAT1 were reduced (pNF-kB: 0.28 versus 0.45 unit/mL, P < 0.05, pSTAT1: 0.16 versus 0.34, P < 0.05), whereas levels of pAkt and pSmad3 were elevated (pAkt: 0.13 versus 0.07, P < 0.01, pSmad3: 1.07 versus 0.47, P < 0.05) in the BM-MSCs group. The levels of pNF-kB, pAkt, and pSmad3 were correlated with aortic diameters. Trophic factors including IGFPB-3, NRF, Activin A and PDGF-AA were secreted from BM-MSCs (IGFBP-3: 35.2 pg/mL, NRF: 3.1 pg/mL, Activin A: 3.1 pg/mL, PDGF-AA: 0.45 pg/mL). Conclusions Our findings suggested that the therapeutic mechanism of BM-MSC–mediated AA regression could contribute to regulation of the NF-kB, Smad3 and Akt signaling pathways. In addition, paracrine actions by factors including NRF, IGFBP-3, Activin A and PDGF-AA might have affected these signaling pathways.

DOI： 10.1016/j.jcyt.2017.07.010

Clarithromycinの大動脈瘤発生抑制効果の作用機序の解明招待有り査読有り

内田亘, 成田裕司, 緒方藍歌, 伊藤英樹, 山名孝治, 寺澤幸枝, 徳田順之, 六鹿雅登, 藤本和朗, 阿部知伸, 大島英揮, 碓氷章彦

日本心臓血管外科学会学術総会抄録集 47回巻頁： 850 - 850 2017年2月

Isolation and characterisation of peripheral blood-derived feline mesenchymal stem cells. 査読有り

Sato K, Yamawaki-Ogata A, Kanemoto I, Usui A, Narita Y

Veterinary journal (London, England : 1997) 216 巻頁： 183 - 8 2016年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Veterinary Journal

The aim of this study was to isolate mesenchymal stem cells (MSCs) from feline peripheral blood (fPB-MSCs) and to characterise the cells’ in vitro properties. The mononuclear cell fractions were isolated from venous blood of cats by density gradient centrifugation and cultured on plastic dishes under various culture conditions to isolate MSCs. When these cells were cultured with 5% autologous plasma (AP) and 10% foetal bovine serum (FBS), adherent spindle shaped fibroblast-like cells (fPB-MSCs) were obtained from 15/22 (68%) cats. These cells were isolated only from medium containing both AP and FBS. The morphology of these MSCs was similar to those isolated from other species and from other feline tissues. fPB-MSCs expanded steadily up to 5–6 passages, but had increased population doubling time during passaging and almost all cells stopped proliferation at passages 7–9. These cells expressed CD44 and CD90, and were mostly negative for major histocompatibility class II and CD4. The cells could be induced to differentiate into adipogenic, osteogenic and chondrogenic cell lineages. These findings indicate that fPB-MSCs can be generated but appear to require specific culture conditions.

DOI： 10.1016/j.tvjl.2016.08.009

大動脈瘤に対する幹細胞療法効果のメカニズムに関して招待有り査読有り

成田裕司, 緒方藍歌, 小坂井基史, 藤本和朗, 六鹿雅登, 阿部知伸, 大島英揮, 碓氷章彦

日本心臓血管外科学会学術総会抄録集 46回巻頁： OP10 - 2 2016年2月

Exosomes Obtained from Mesenchymal Stem Cells as a Novel Therapeutic Tool for Aortic Aneurysm 招待有り査読有り

Yamawaki-ogata, A; Kosakai, M; Usui, A; Narita, Y

TISSUE ENGINEERING PART A 21 巻頁： S211 - S212 2015年9月

Skin-derived precursor cells promote wound healing in diabetic mice. 査読有り

Sato H, Ebisawa K, Takanari K, Yagi S, Toriyama K, Yamawaki-Ogata A, Kamei Y

Annals of plastic surgery 74 巻 ( 1 ) 頁： 114 - 20 2015年1月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Annals of Plastic Surgery

Background: Impaired wound healing as one of the complications arising from diabetes mellitus is a serious clinical issue. Recently, various cell therapies have been reported for promotion of wound healing. Skin-derived precursor cells (SKPs) are multipotent adult stem cells with the tendency to differentiate into neurons. We investigated the potency of promoting diabetic wound healing by the application of SKPs. Methods: Skin-derived precursor cells isolated from diabetic murine skin were cultured in sphere formation medium. At passage 2, they were suspended in phosphate-buffered saline (PBS), and applied topically to full-thickness excisional cutaneous wounds in diabetic mice. Application of PBS served as controls (n = 21 for each group; n = 42 total). Time to closure and percentage closure were calculated by morphometry. Wounds were harvested at 10 and 28 days and then processed, sectioned, and stained (CD31, α-smooth muscle actin, and neurofilament heavy chain) to quantify vascularity and neurofilaments. Results: Wounds treated with SKPs demonstrated a significantly decreased time to closure (18.63 days) compared with PBS-control wounds (21.72 days, P < 0.01), and a significant improvement in percentage closure at 7, 10, 14, and 18 days compared with PBS-control wounds (P < 0.01). Histological analysis showed that the Capillary Score (the number of vessels/mm2) was significantly higher in SKP-treated wounds at day 10 but not at day 28. Nerve Density (the number of neurofilaments/mm2) had increased significantly in SKP-treated wounds at day 28 compared with control group. Some applied SKPs were stained by neurofilament heavy chain, which demonstrates that SKPs directly differentiated into neurons. Conclusions: Skin-derived precursor cells promoted diabetic wound healings through vasculogenesis at the early stage of wound healing. Skin-derived precursor cells are a possible therapeutic tool for diabetic impaired wound healing.

DOI： 10.1097/SAP.0000000000000342

Mesenchymal stem cells for treatment of aortic aneurysms. 査読有り

Yamawaki-Ogata A, Hashizume R, Fu XM, Usui A, Narita Y

World journal of stem cells 6 巻 ( 3 ) 頁： 278 - 87 2014年7月

Therapeutic potential of bone marrow-derived mesenchymal stem cells in formed aortic aneurysms of a mouse model. 査読有り

Yamawaki-Ogata A, Fu X, Hashizume R, Fujimoto KL, Araki Y, Oshima H, Narita Y, Usui A

European journal of cardio-thoracic surgery : official journal of the European Association for Cardio-thoracic Surgery 45 巻 ( 5 ) 頁： e156 - 65 2014年5月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：European Journal of Cardio-thoracic Surgery

OBJECTIVES: An aortic aneurysm (AA) is caused by atherosclerosis with chronic inflammation. Mesenchymal stem cells (MSCs) have potential anti-inflammatory properties. In this study, we examined whether an already-formed AA can be treated by intravenous injection of bone marrow-derived (BM)-MSCs in a mouse model. METHODS: AA was induced in apolipoprotein E-deficient mice by angiotensin II-infusion for 28 days through sub-cutaneous osmotic mini-pumps. After that, 1 × 106 BM-MSCs (in 0.2 ml saline) or 0.2 ml saline as a control was injected via the tail vein. Mice were sacrificed at 2 (saline group n = 10, BM-MSC group n = 10), 4 (saline group n = 6, BM-MSC group n = 7) or 8 weeks (saline group n = 5, BM-MSC group n = 6) after injection. The aortic tissues of each group were dissected. Aortic diameter, elastin content, matrix metalloproteinase (MMP)-2 and -9 enzymatic activity and cytokine concentrations were measured, as was macrophage infiltration, which was also evaluated histologically. RESULTS: The incidence of AA in the BM-MSC group was reduced at 2 weeks (BM-MSC 40% vs saline 100%, P < 0.05), and aortic diameter was reduced at 2 and 4 weeks (2 weeks: 1.40 vs 2.29 mm, P < 0.001; 4 weeks: 1.73 vs 2.32 mm, P < 0.05). The enzymatic activities of MMP-2 and -9 were reduced in the BM-MSC group at 2 weeks (active-MMP-2: 0.28 vs 0.45 unit/ml, P < 0.05; active-MMP-9: 0.16 vs 0.34 unit/ml, P < 0.05). Inflammatory cytokines were down-regulated in the BM-MSC group (interleukin-6: 2 weeks: 1475.6 vs 3399.5 pg/ml, P < 0.05; 4 weeks: 2184.7 vs 3712.8 pg/ml, P < 0.05 and monocyte chemotactic protein-1: 2 weeks: 208.0 vs 352.7 pg/ml, P < 0.05) and insulin-like growth factor (IGF)-1 and tissue inhibitor of metalloproteinase (TIMP)-2 were up-regulated in the BM-MSC group at 2 weeks (IGF-1: 4.7 vs 2.0 ng/ml, P < 0.05; TIMP-2: 9.5 vs 4.0 ng/ml, P < 0.001). BM-MSC injection inhibited infiltration of M1 macrophages and preserved the construction of elastin. CONCLUSIONS: Our results suggest that BM-MSCs might be an effective treatment for AA. Further investigation is necessary to optimize the injected dosage and the frequency of BM-MSCs to prevent a transient effect. © The Author 2014. Published by Oxford University Press on behalf of the European Association for Cardio-Thoracic Surgery. All rights reserved.

DOI： 10.1093/ejcts/ezu018

Intravenous administration of mesenchymal stem cells prevents angiotensin II-induced aortic aneurysm formation in apolipoprotein E-deficient mouse. 査読有り

大動脈瘤に対する幹細胞療法投与法による大動脈瘤進展抑制効果の違い招待有り査読有り

成田裕司, 緒方藍歌, 符顕明, 荒木善盛, 阿部知伸, 大島英揮, 碓氷章彦

日本心臓血管外科学会雑誌 43 巻 ( Suppl. ) 頁： 263 - 263 2014年1月

Fu XM, Yamawaki-Ogata A, Oshima H, Ueda Y, Usui A, Narita Y

Journal of translational medicine 11 巻 ( 1 ) 頁： 175 2013年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Translational Medicine

Background: Mesenchymal stem cells (MSCs) are known to be capable of suppressing inflammatory responses. We previously reported that intra-abdominal implantation of bone marrow-derived MSCs (BM-MSCs) sheet by laparotomy attenuated angiotensin II (AngII)-induced aortic aneurysm (AA) growth in apolipoprotein E-deficient (apoE-/-) mice through anti-inflammation effects. However, cell delivery by laparotomy is invasive; we here demonstrated the effects of multiple intravenous administrations of BM-MSCs on AngII-induced AA formation.Methods: BM-MSCs were isolated from femurs and tibiae of male apoE-/- mice. Experimental AA was induced by AngII infusion for 28 days in apoE-/- mice. Mice received weekly intravenous administration of BM-MSCs (n=12) or saline (n=10). After 4 weeks, AA formation incidence, aortic diameter, macrophage accumulation, matrix metalloproteinase (MMP)' activity, elastin content, and cytokines were evaluated.Results: AngII induced AA formation in 100% of the mice in the saline group and 50% in the BM-MSCs treatment group (P < 0.05). A significant decrease of aortic diameter was observed in the BM-MSCs treatment group at ascending and infrarenal levels, which was associated with decreased macrophage infiltration and suppressed activities of MMP-2 and MMP-9 in aortic tissues, as well as a preservation of elastin content of aortic tissues. In addition, interleukin (IL)-1β, IL-6, and monocyte chemotactic protein-1 significantly decreased while insulin-like growth factor-1 and tissue inhibitor of metalloproteinases-2 increased in the aortic tissues of BM-MSCs treatment group.Conclusions: Multiple intravenous administrations of BM-MSCs attenuated the development of AngII-induced AA in apoE-/- mice and may become a promising alternative therapeutic strategy for AA progression. © 2013 Fu et al.; licensee BioMed Central Ltd.

DOI： 10.1186/1479-5876-11-175

Prevention of arterial graft spasm in rats using a vasodilator-eluting biodegradable nano-scaled fibre. 査読有り

Yagami K, Yamawaki-Ogata A, Satake M, Kaneko H, Oshima H, Usui A, Ueda Y, Narita Y

Interactive cardiovascular and thoracic surgery 17 巻 ( 1 ) 頁： 16 - 22 2013年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Interactive Cardiovascular and Thoracic Surgery

OBJECTIVESArterial graft spasm occasionally causes circulatory collapse immediately following coronary artery bypass graft. The aim of this study is to evaluate the efficacy of our developed materials, which were composed of milrinone (phosphodiesterase III inhibitor) or diltiazem (calcium-channel blocker), with nano-scaled fibre made of biodegradable polymer to prevent arterial spasm.METHODSMilrinone- or diltiazem-releasing biodegradable nano-scaled fibres were fabricated by an electrospinning procedure. In vivo milrinone- or diltiazem-releasing tests were performed to confirm the sustained release of the drugs. An in vivo arterial spasm model was established by subcutaneous injection of noradrenalin around the rat femoral artery. Rats were randomly divided into four groups as follows: those that received 5 mg of milrinone-releasing biodegradable nano-scaled fibre (group M, n = 14); 5 mg of diltiazem-releasing biodegradable nano-scaled fibre (group D, n = 12); or those that received fibre without drugs (as a control; group C, n = 14) implanted into the rat femoral artery. In the fourth group, sham operation was performed (group S, n = 10). One day after the implantation, noradrenalin was injected in all groups. The femoral arterial blood flow was measured continuously before and after noradrenalin injection. The maximum blood flow before noradrenalin injection and minimum blood flow after noradrenalin injection were measured.RESULTSIn vivo drug-releasing test revealed that milrinone-releasing biodegradable nano-scaled fibre released 78% of milrinone and diltiazem-releasing biodegradable nano-scaled fibre released 50% diltiazem on the first day. The ratios of rat femoral artery blood flow after/before noradrenalin injection in groups M (0.74 ± 0.16) and D (0.72 ± 0.05) were significantly higher than those of groups C (0.54 ± 0.09) and S (0.55 ± 0.16) (P < 0.05).CONCLUSIONNoradrenalin-induced rat femoral artery spasm was inhibited by the implantation of milrinone-releasing biodegradable nano-scaled fibre or diltiazem-releasing biodegradable nano-scaled fibre. These results suggested that our materials might be effective for the prevention of arterial graft spasm after coronary artery bypass graft. © 2013 The Author. Published by Oxford University Press on behalf of the European Association for Cardio-Thoracic Surgery. All rights reserved.

DOI： 10.1093/icvts/ivt092

Long-term results of tissue-engineered small-caliber vascular grafts in a rat carotid arterial replacement model 査読有り

KUWABARA Fumiaki, NARITA Yuji, YAMAWAKI OGATA Aika, SATAKE Makoto, KANEKO Hiroaki, OSHIMA Hideki, USUI Akihiko, UEDA Yuichi

15 巻 ( 4 ) 頁： 399 - 405 2012年12月

Long-term results of tissue-engineered small-caliber vascular grafts in a rat carotid arterial replacement model. 査読有り

Kuwabara F, Narita Y, Yamawaki-Ogata A, Satake M, Kaneko H, Oshima H, Usui A, Ueda Y

Journal of artificial organs : the official journal of the Japanese Society for Artificial Organs 15 巻 ( 4 ) 頁： 399 - 405 2012年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Artificial Organs

The concept of tissue engineered small-caliber vascular grafts (TE-SCVGs) is theoretically ideal. In this study, we evaluated the long-term (more than 1 year) course of TE-SCVGs using a rat carotid arterial replacement model. We fabricated a TE-SCVG scaffold (0.7 mm in diameter) with electrospun nano-scale fibers. Poly-ε-caprolactone was used as a biodegradable polymer. These artificial vessels were then used in carotid arterial replacement performed on Sprague-Dawley rats. The implanted grafts were removed at an early phase (1, 2, 6 weeks), middle phase (12, 24 weeks), and late phase (48, 72 weeks) after implantation. Twenty-nine patent grafts from among the 40 implanted grafts (patency 72.5 %) could be evaluated. No aneurysm formation was observed during the follow-up period. Endothelial cells positive for immunostaining with von Willebrand factor were found to be already attached to the inner surface of the TE-SCVGs in the early phase. The percentage of smooth muscle cell specific marker (α-smooth muscle actin and calponin with fluorescent immunostaining) positive cells, which seemed to be mesenchymal cells in the graft wall, increased with time, while, in contrast, the scaffold material decreased. Even after 72 weeks, however, although the scaffold material had degraded, it had not disappeared completely. These results show that the novel TE-SCVGs we developed were still functioning in the rat carotid arterial circulation after more than 1 year. However, further investigations will be required with regard to regeneration of the SMC layer and the complete degradation of graft materials. © 2012 The Japanese Society for Artificial Organs.

DOI： 10.1007/s10047-012-0652-6

PS-106-2 動脈グラフと攣縮予防のための血管拡張薬徐放生体吸収性ナノファイバーの開発(PS-106 心血管基礎,ポスターセッション,第112回日本外科学会定期学術集会) 査読有り

Development of novel statin releasing biodegradable nano-scaled fiber for bone tissue engineering 招待有り査読有り

Wadagaki, R; Mizuno, D; Yamawaki-Ogata, A; Fu, X; Satake, M; Kaneko, H; Ueda, M; Narita, Y

JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE 6 巻頁： 329 - 329 2012年9月

Possibility of cell therapy for aortic aneurysm 招待有り査読有り

Yamawaki-Ogata, A; Fu, X; Hashizume, R; Oshima, H; Usui, A; Narita, Y

JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE 6 巻頁： 149 - 149 2012年9月

八神啓, 成田裕司, 緒方藍歌, 佐竹真, 兼子博章, 大島英揮, 碓氷章彦, 上田裕一

日本外科学会雑誌 113 巻 ( 2 ) 頁： 703 2012年3月

Effect of GDF-5 and BMP-2 on the expression of tendo/ligamentogenesis-related markers in human PDL-derived cells. 招待有り査読有り

Inoue M, Ebisawa K, Itaya T, Sugito T, Yamawaki-Ogata A, Sumita Y, Wadagaki R, Narita Y, Agata H, Kagami H, Ueda M

Oral diseases 18 巻 ( 2 ) 頁： 206 - 12 2012年3月

担当区分：筆頭著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Oral Diseases

Objectives: The effect of growth differentiation factor 5 and bone morphogenetic protein 2 on human periodontal ligament-derived cells was investigated with special reference to tendo/ligamentogenesis-related markers. Materials and Methods: Effects of each factor were analyzed by quantitative PCR for scleraxis and tenomodulin and by western blotting for scleraxis. After exposure to those factors, STRO-1-positive and STRO-1-negative fractions of human periodontal ligament tissues were isolated with an immunomagnetic cell sorting system, and the expression of scleraxis in each fraction was analyzed by western blotting. Non-separated crude cells were used as a control. Results: Growth differentiation factor 5 and bone morphogenetic protein 2 did not increase alkaline phosphatase activity in crude periodontal ligament-derived cells. Growth differentiation factor 5, but not bone morphogenetic protein 2, increased the expression of scleraxis in crude, STRO-1-positive and STRO-1-negative periodontal ligament-derived cells. The expression of scleraxis in STRO-1-positive periodontal ligament-derived cells was significantly less compared to that in crude P2 and STRO-1-negative periodontal ligament-derived cells. Conclusion: Growth differentiation factor 5 induced the expression of scleraxis and may enhance tendo/ligamentogenesis in human periodontal ligament-derived cells. The expression of scleraxis was higher in STRO-1-negative fraction, suggesting more differentiated state of the cells. © 2011 John Wiley & Sons A/S.

DOI： 10.1111/j.1601-0825.2011.01871.x