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Increased osteogenesis with hydroxyapatite constructs combined with serially-passaged bone marrow-derived mesenchymal stem cells

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Stem Cell Discovery 2012年第4期2卷 133-140页

作者： Manabu Akahane Tomoyuki Ueha Takamasa Shimizu Yusuke Inagaki akira kido Tomoaki Imamura Kenji Kawate Yasuhito Tanaka Department of Artificial Joint and Regenerative Medicine Nara Medical University Nara Japan Department of Orthopedic Surgery Nara Medical University Nara Japan Department of Public Health Health Management and Policy Nara Medical University School of Medicine Nara Japan

We have previously reported on both the osteogenic potential of hydroxyapatite (HA) combined with bone marrow-derived mesenchymal stem cells (BMSCs) and a method involving osteogenic matrix cell sheet transplantation of BMSCs. In the present study, we assessed the osteogenic potential of serially-passaged BMSCs, both in vitro and in vivo. We also assessed whether an additional cell-loading technique can regain the osteogenic potential of the constructs combined with serially-passaged BMSCs. The present study revealed that passage (P) 1 cells cultured in osteogenic-induced medium showed strong positive staining for alkaline phosphatase (ALP) and Alizarin Red S, whereas P3 cells showed faint staining for ALP, with no Alizarin Red S staining. Staining of P1, P2 and P3 cells were progressively weaker, indicating that the osteogenic potential of the serially-passaged rat BMSCs is lost after P3 in vitro. The in vivo study showed that little bone formation was observed in the HA constructs seeded with P3 cells, 4 weeks after subcutaneous implantation. However, P3 cell/HA constructs which had increased cell-loading showed abundant bone formation within the pores of the HA construct. ALP and osteocalcin mRNA expression in these constructs was significantly higher than that of constructs with regular cell-seeding. The present study indicates that the osteogenic potential of the constructs with serially-passaged BMSCs is increased by additional cell-loading. This method can be applied to cases requiring hard tissue reconstruction, where BMSCs require serial expansion of cells.

关键词： Osteogenesis Mesenchymal Stem Cells Serial Passaging Hydroxyapatite Tissue Engineering Bone Reconstruction

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Utility of tricalcium phosphate and osteogenic matrix cell sheet constructs for bone defect reconstruction

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World Journal of Stem Cells 2015年第5期7卷 873-882页

作者： Tomoyuki Ueha Manabu Akahane Takamasa Shimizu Yoshinobu Uchihara Yusuke Morita Naoya Nitta akira kido Yusuke Inagaki Kenji Kawate Yasuhito Tanaka Department of Orthopedic Surgery Nara Medical University Department of Public Health Health Management and Policy Nara Medical University Faculty of Medicine Department of Biomedical Engineering Doshisha University Department of Arthroplasty and Regenerative Medicine Nara Medical University

AIM: To determine the effects of transplanting osteogenic matrix cell sheets and beta-tricalcium phosphate(TCP) constructs on bone formation in bone ***: Osteogenic matrix cell sheets were prepared from bone marrow stromal cells(BMSCs), and a porous TCP ceramic was used as a scaffold. Three experimental groups were prepared, comprised of TCP scaffolds(1) seeded with BMSCs;(2) wrapped with osteogenic matrix cell sheets; or(3) both. Constructs were implanted into a femoral defect model in rats and bone growth was evaluated by radiography, histology, biochemistry, and mechanical testing after 8 wk. RESULTS: In bone defects, constructs implanted with cell sheets showed callus formation with segmentalor continuous bone formation at 8 wk, in contrast to TCP seeded with BMSCs, which resulted in bone nonunion. Wrapping TCP constructs with osteogenic matrix cell sheets increased their osteogenic potential and resulting bone formation, compared with conventional bone tissue engineering TCP scaffolds seeded with BMSCs. The compressive stiffness(mean ± SD) values were 225.0 ± 95.7, 30.0 ± 11.5, and 26.3 ± 10.6 MPa for BMSC/TCP/Sheet constructs with continuous bone formation, BMSC/TCP/Sheet constructs with segmental bone formation, and BMSC/TCP constructs, respectively. The compressive stiffness of BMSC/TCP/Sheet constructs with continuous bone formation was significantly higher than those with segmental bone formation and BMSC/TCP ***: This technique is an improvement over current methods, such as TCP substitution, and is useful for hard tissue reconstruction and inducing earlier bone union in defects.

关键词： Bone marrow stromal cells Osteogenesis Bone regeneration Tissue engineering Calcium phosphate

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Myocardial Segmentation of Area at Risk Based on Coronary Computed Tomography Angiography and Voronoi Diagram in Comparison with Magnetic Resonance Perfusion Imaging

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Open Journal of Radiology 2017年第1期7卷 9-22页

作者： Naoki Fukuyama Teruhito kido akira Kurata Yuki Tanabe Tomoyuki kido Takahiro Yokoi Ryo Ogawa Hikaru Nishiyama Teruyoshi Uetani Teruhito Mochizuki Department of Radiology Ehime University Graduate School of Medicine Toon Japan Department of Cardiology Ehime University Graduate School of Medicine Toon Japan

Purpose: To assess the clinical feasibility of automated segmentation of the myocardial area at risk (MAAR) using coronary computed tomography angiography (CT-MAAR), as compared to stress magnetic resonance myocardial perfusion imaging (MR-MPI). Materials and Methods: Thirty patients who underwent coronary computed tomography angiography (CTA) and stress MR-MPI were retrospectively evaluated. The myocardial territory of the left ventricle (LV) distal to coronary artery stenosis (≥50% or ≥70% stenosis on coronary CTA) was three-dimensionally quantified using a Voronoi diagram. The ratio of all stenosis-related territories to the LV volume was defined as CT-MAAR (%-LV volume). The proportion of segments with perfusion defects in stress MR-MPI to the total of 16 segments (range: 0% - 100%;with a 6.3%-interval scale) was defined as the reference. Correlation was assessed using Spearman’s test. The capability of CT-MAAR to predict the ischemic burden was assessed. Results: Stress MR-MPI depicted a median ischemic burden of 25.2% (range: 18.9% - 44.1%) in 30 patients without myocardial infarction. When CTA stenosis criteria of ≥50% (n = 30) and ≥70% (n = 27) were applied to estimate CT-MAAR, the median CT-MAAR values were 48.2% (31.6% - 64.3%) and 32.5% (23.7% - 51.9%), respectively. The correlations between the CT-MAAR values and the MR-based ischemic burden were significant (0.73 and 0.97 for ≥50% and ≥70% stenosis, respectively). CT-MAAR predicted the MR-based ischemic burden within ±1 segment of %-LV (6.3%) in 40% (12/30) of patients with ≥50% stenosis, and in 81.5% (22/27) of patients with ≥70% stenosis. Conclusions: Comprehensive assessment of resting coronary CTA combined with Voronoi diagram-based myocardial segmentation may help predict the myocardial ischemic burden in patients with severe coronary CTA stenosis.

关键词： Computed Tomography Ischemia Myocardial Area at Risk

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Effect of SRD-P401 on the Discomfort of Neck and/or Shoulder Stiffness

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American Journal of Plant Sciences 2017年第5期8卷 1065-1076页

作者： akira Iizuka Maasa Sonoda Ruka Goto Tokunori Matsui Chiaki Kudoh Katsumi kido Yasuhiro Komatsu Center for Supporting Pharmaceutical Education Daiich University of Pharmacy Fukuoka Japan Clinical Pharmacy Education Center Japan College of Pharmaceutical Science Saitama Japan Sun R and D Institute for Natural Medicines Co. Inc. Tokyo Japan KUDOH CHIAKI Clinic for Neurosurgery & Neurology Tokyo Brain & Spinal Cord Research Institute Tokyo Japan Department of Pharmacognosy Daiich University of Pharmacy Fukuoka Japan

SRD-P401 is a newly developed healthy food supplement which has an anti-inflammatory and analgesic effect. In this report, the effects of SRD-P401 on neck and shoulder stiffness were assessed by an inquiry method in the public field. The stiffness symptoms of all participants were improved after taking SRD-P401. The symptoms of the participants who took 3 g/day and/or 9 g/day of SRD-P401 improved in a week after taking it. Unpleasant adverse effects in stomach and alimentally organs were not found during the test period. SRD-P401 showed the anti-nociceptive effect on the acetic-acid-induces abdominal writhing in rats. The potency was equivalent to aspirin. Moreover, to investigate the ability of the anti-inflammatory effects of SRD-P401, we exerted by inhibiting the Cyclooxygenase (COX) enzyme activity. SRD-P401 and its constituents inhibit the COX activity. The results of this field test indicated SRD-P401 would be a beneficial and useful healthy food supplement for a person with stiffness in neck and/or shoulder.

关键词： Supplement Anti-Inflammatory Anti-Nociceptive SRD-P401

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Evaluation of the Estrogenic Activity of Pueraria (Kudzu) Flower Extract and Its Major Isoflavones Using ER-Binding and Uterotrophic Bioassays

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Pharmacology & Pharmacy 2013年第2期4卷 255-260页

作者： Tomoyasu Kamiya akira Takano Yayoi kido Yuki Matsuzuka Mayu Sameshima-Kamiya Masahito Tsubata Motoya Ikeguchi Kinya Takagaki Junei Kinjo Faculty of Pharmaceutical Sciences Fukuoka University Fukuoka Japan Research and Development Division Toyoshinyaku Co. Ltd. Tosu-shi Japan

Pueraria flower extract (PFE) is a hot water extract of the Kudzu flower (Pueraria thomsonii). Tea made from dried Kudzu flower is widely used inChina, and PFE is utilized as a nutritional supplement inJapan. PFE contains unique isoflavones such as 6-hydroxygenistein 6,7-di-O-glucoside (6HGDG), tectorigenin 7-O-xylosylglucoside (TGXG), and tectoridin. 6HGDG is known to be metabolized into 6-hydroxygenistein, and TGXG and tectoridin are known to be metabolized into tectorigenin in the digestive tract. Isoflavones typically mimic the effects β-estradiol has on estrogen receptors (ERs) and may influence the female genital system in the case of excessive intake. As a result, the upper limit of safe daily consumption of soy isoflavones has been enforced inJapan. In the present study, ER-binding assays were performed using the EnBio estrogen receptor/cofactor assay system to compare the estrogenic activity of 6-hydroxygenistein and tectorigenin to that of the soy isoflavone genistein. In addition, uterotrophic bioassays were performed to investigate the estrogenic effects of PFE in vivo. The ER-binding assays revealed that the ER-binding affinities of 6-hydroxygenistein and tectorigenin were approximately 0.01 - 0.04 that of genistein. Soy isoflavone products also induced an increase in uterine wet and blotted weight at doses of 500 mg/kg and 1000 mg/kg, whereas PFE did not cause adverse estrogenic effects, even at a dose of 1000 mg/kg. Based on these results, PFE does not appear to contain compounds with strong estrogenic activity or cause adverse estrogenic effects in vivo. Importantly, the results of this study confirm the safety of PFE as a food supplement.

关键词： Estrogenic Activity 6-Hydroxygenistein Kudzu Flower Extract Safety Evaluation Tectorigenin

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