2025/06/23 更新

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写真a

ヤマシタ ダイキ
山下 大輝
YAMASHITA DAIKI
所属
九州大学病院 歯内治療科 助教
歯学府 歯学専攻(併任)
歯学部 歯学科(併任)
職名
助教
連絡先
メールアドレス

研究分野

  • ライフサイエンス / 保存治療系歯学

経歴

  • 九州大学  助教 

学歴

  • 九州大学    

論文

  • Establishment of Periodontal Ligament Stem Cell-like Cells Derived from Feeder-Free Cultured Induced Pluripotent Stem Cells

    Yamashita, D; Hamano, S; Hasegawa, D; Sugii, H; Itoyama, T; Ikeya, M; Maeda, H

    STEM CELLS AND DEVELOPMENT   33 ( 23-24 )   665 - 676   2024年12月   ISSN:1547-3287 eISSN:1557-8534

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    記述言語:英語   出版者・発行元:Stem Cells and Development  

    The periodontal ligament (PDL) is a fibrous connective tissue that connects the cementum of the root to the alveolar bone. PDL stem cells (PDLSCs) contained in the PDL can differentiate into cementoblasts, osteoblasts, and PDL fibroblasts, with essential roles in periodontal tissue regeneration. Therefore, PDLSCs are expected to be useful in periodontal tissue regeneration therapy. In a previous study, we differentiated induced pluripotent stem cells (iPSCs) into PDLSC-like cells (iPDLSCs), which expressed PDL-related markers and mesenchymal stem cell (MSC) markers; they also exhibited high proliferation and multipotency. However, the iPSCs used in this differentiation method were cultured on mouse embryonic fibroblasts; thus, they constituted on-feeder iPSCs (OF-iPSCs). Considering the risk of contamination with feeder cell-derived components, iPDLSCs differentiated from OF-iPSCs (ie, OF-iPDLSCs) are unsuitable for clinical applications. In this study, we aimed to obtain PDLSC-like cells from feeder-free iPSCs (FF-iPSCs) using OF-iPDLSC differentiation method. First, we differentiated FF-iPSCs into neural crest cell-like cells (FF-iNCCs) and confirmed that FF-iNCCs expressed NCC markers (eg, Nestin and p75NTR). Then, we cultured FF-iNCCs on human primary PDL cell-derived extracellular matrix for 2 weeks; the resulting cells were named FF-iPDLSCs. FF-iPDLSCs exhibited higher expression of PDL-related and MSC markers compared with OF-iPDLSCs. FF-iPDLSCs also demonstrated proliferation and multipotency in vitro. Finally, we analyzed the ability of FF-iPDLSCs to form periodontal tissue in vivo upon subcutaneous transplantation with β-tricalcium phosphate scaffolds into dorsal tissues of immunodeficient mice. Eight weeks after transplantation, FF-iPDLSCs had formed osteocalcin-positive bone/cementum-like tissues and collagen 1-positive PDL-like fibers. These results suggested that we successfully obtained PDLSC-like cells from FF-iPSCs. Our findings will contribute to the development of novel periodontal regeneration therapies.

    DOI: 10.1089/scd.2024.0122

    Web of Science

    Scopus

    PubMed

  • Effect of Fibrillin-2 on Differentiation into Periodontal Ligament Stem Cell-Like Cells Derived from Human-Induced Pluripotent Stem Cells

    Hamano, S; Yamashita, D; Hasegawa, D; Sugii, H; Itoyama, T; Maeda, H

    STEM CELLS AND DEVELOPMENT   33 ( 9-10 )   228 - 238   2024年5月   ISSN:1547-3287 eISSN:1557-8534

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    記述言語:英語   出版者・発行元:Stem Cells and Development  

    Periodontal tissue regeneration is important for preserving teeth. Periodontal ligament stem cells (PDLSCs) are useful in periodontal tissue regeneration; however, tooth extraction is required to obtain these cells. Therefore, we focused on induced pluripotent stem (iPS) cells and established a method to obtain PDLSC-like cells from iPS cells. Specifically, we first differentiated iPS cells into neural crest-like cells (iNCs). Next, we obtained PDLSC-like cells (iPDLSCs) by culturing iNCs on extracellular matrix (ECM) derived from human primary periodontal ligament cells (HPDLCs). This differentiation method suggested that ECM derived from HPDLCs is important for iPDLSC differentiation. Thus, we aimed to identify the PDLSC-inducing factor present in HPDLC-derived ECM in this study. We first performed comprehensive analyses of HPDLC genes and identified fibrillin-2 (FBN2), an ECM-related factor. Furthermore, to clarify the effect of FBN2 on iPDLSC differentiation, we cultured iNCs using ECM derived from HPDLCs with FBN2 knocked down. As a result, expression of PDL-related markers was reduced in iNCs cultured on ECM derived from HPDLCs transfected with FBN2 siRNA (iNC-siFBN2) compared with iPDLSCs. Furthermore, the expression of CD105 (a mesenchymal stem cell marker), proliferation ability, and multipotency of iNC-siFBN2 were lower compared with iPDLSCs. Next, we cultured iNCs on FBN2 recombinant protein; however, expression of PDL-related markers did not increase compared with iPDLSC. The present results suggest the critical involvement of FBN2 in inducing iPDLSCs from iNCs when in fact it does not promote iPDLSC differantiation. Therefore, we need to elucidate the entire HPDLC-ECMs, responsible for iPDLSCs induction.

    DOI: 10.1089/scd.2024.0013

    Web of Science

    Scopus

    PubMed

  • 根尖孔穿通への熱処理ニッケルチタンファイルとTri Auto ZX2新駆動方式の応用

    山下 大輝, 友清 淳, 濱野 さゆり, 前田 英史

    日本歯内療法学会雑誌   45 ( 1 )   34 - 40   2024年1月   ISSN:1347-8672

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    記述言語:日本語   出版者・発行元:(一社)日本歯内療法学会  

    ニッケルチタン(NiTi)製ファイルは根管治療を行う上で重要な器具のひとつである。NiTi合金は熱処理によって圧縮強さや引っ張り強さが変化することが知られており、プログライダーは熱処理加工されていないNiTiファイルと比較して柔軟性と破折抵抗性に優れる。本研究では、プログライダーとOptimum Glide Path(OGP)モードならびに新駆動方式(N-OGPモード)を組み合わせ、穿通の可否および穿通までに要した時間を検証することで、レシプロケーション運動による熱処理NiTiファイルの使用が根尖孔穿通のオプションのひとつとなる可能性について検討した。根管の先端径が0.1mmであるS字根管模型70本を実験に使用し、術者は当科に所属する臨床経験年数6~11年の歯科医師7名(Practician Group)と経験1年未満の研修歯科医7名(Trainee Group)とした。Practician Groupではすべての術者において5群とも穿通が可能であり、穿通率は100%であった。穿通までに要した時間は短い順にN-OGP+SPF、OGP+SPF、OGP+PRG、N-OGP+PRG、KFであった。Trainee Groupでは1術者のOGP+PRGにおいてのみファイル破折が生じたため穿通不可であったが、他の術者および実験群では穿通が可能であり、穿通率は97.1%であった。穿通までに要した時間は短い順にOGP+SPF、N-OGP+PRG、N-OGP+SPF、OGP+PRG、KFであった。

  • 根尖孔穿通への熱処理ニッケルチタンファイルとTri Auto ZX2新駆動方式の応用

    山下 大輝, 友清 淳, 濱野 さゆり, 前田 英史

    日本歯内療法学会雑誌   45 ( 1 )   34 - 40   2024年   ISSN:13478672 eISSN:24239429

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    記述言語:日本語   出版者・発行元:一般社団法人 日本歯内療法学会  

    <p><b>Abstract : </b>Purpose : The aim of this study was to evaluate the efficacy of heat-treated nickel-titanium (NiTi) files and an endodontic motor with reciprocating motion on the establishment of apical patency.</p><p> Materials and Methods : Seventy S-shaped root canal blocks were used in this study. Dentists with 6 to 11 years of clinical experience (Practician Group, n=7) and trainee dentists with 1 year of clinical experience (Trainee Group, n=7) tried to establish apical patency using the following five techniques. Group 1 : Manual operation with stainless steel (SS) files (#15/.02 K file). Group 2 : Tri Auto ZX2 Optimum Glide Path (OGP) mode with SS files (#15/.02 Superfile [SPF] ; OGP+SPF). Group 3 : Tri Auto ZX2 OGP mode with heat-treated NiTi files (#16/.02-.08 ProGlider [PRG] ; OGP+PRG). Group 4 : Tri Auto ZX2 new OGP (N-OGP) mode with Superfile (N-OGP+SPF). Group 5 : Tri Auto ZX2 N-OGP mode with ProGlider (N-OGP+PRG). The time required to establish apical patency was measured. The surface of the K file, Superfile, and ProGlider before and after the establishment of apical patency was observed by scanning electron microscope.</p><p> Results and Discussion : The rate of establishment of apical patency was 100% and 97.1% in the Practician Group and Trainee Group, respectively. One file fracture was observed in OGP+PRG in the Trainee Group. KF required significantly more time than the other four groups in the Practician Group. This tendency was also observed in the Trainee Group, however there was no significant difference among the five groups. No significant difference was observed between the Practician Group and Trainee Group in the five methods. No morphological change was observed on the surface of the K file, Superfile, and ProGlider after the establishment of apical patency.</p><p> Conclusion : N-OGP+PRG is a new treatment option that enables apical patency to be established in a shorter time than manual operation with a K file.</p>

    DOI: 10.20817/jeajournal.45.1_34

    CiNii Research

  • Dopamine is involved in reparative dentin formation through odontoblastic differentiation of dental pulp stem cells

    Fujino, S; Hamano, S; Tomokiyo, A; Sugiura, R; Yamashita, D; Hasegawa, D; Sugii, H; Fujii, S; Itoyama, T; Miyaji, H; Maeda, H

    SCIENTIFIC REPORTS   13 ( 1 )   5668   2023年4月   ISSN:2045-2322

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    記述言語:英語   出版者・発行元:Scientific Reports  

    Conventional direct pulp-capping materials induce pulp cells to secrete various biomolecules in pulp tissues that promote reparative dentin formation through induction of odontoblastic differentiation of dental pulp stem cells (DPSCs). However, these biomolecules sometimes induce bone-like dentin with poor sealing properties. Therefore, exploration of biomolecules that allow tight sealing by tubular reparative dentin is required. We recently reported that dopamine (DA) is involved in dentinogenesis. Hence, we investigated the effect of DA on odontoblastic differentiation of DPSCs and reparative dentin formation. Both tyrosine hydroxylase (TH), a DA synthetase, and DA were expressed in odontoblast-like cells in vivo. In vitro, their expression was increased during odontoblastic differentiation of DPSCs. Furthermore, TH-overexpressing DPSCs had promoted odontoblastic differentiation and DA production. Moreover, DA stimulation promoted their differentiation and induced tubular reparative dentin. These results suggest that DA produced by TH is involved in odontoblastic differentiation of DPSCs and has an inductive capacity for reparative dentin formation similar to primary dentin. This study may lead to the development of therapy to preserve vital pulp tissues.

    DOI: 10.1038/s41598-023-32126-1

    Web of Science

    Scopus

    PubMed

  • Current Application of iPS Cells in the Dental Tissue Regeneration

    Hamano, S; Sugiura, R; Yamashita, D; Tomokiyo, A; Hasegawa, D; Maeda, H

    BIOMEDICINES   10 ( 12 )   2022年12月   ISSN:2227-9059 eISSN:2227-9059

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  • PAX9 Is Involved in Periodontal Ligament Stem Cell-like Differentiation of Human-Induced Pluripotent Stem Cells by Regulating Extracellular Matrix

    Sugiura, R; Hamano, S; Tomokiyo, A; Hasegawa, D; Yoshida, S; Sugii, H; Fujino, S; Adachi, O; Kadowaki, M; Yamashita, D; Maeda, H

    BIOMEDICINES   10 ( 10 )   2022年10月   ISSN:2227-9059 eISSN:2227-9059

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    記述言語:英語   出版者・発行元:Biomedicines  

    Periodontal ligament stem cells (PDLSCs) play central roles in periodontal ligament (PDL) tissue homeostasis, repair, and regeneration. Previously, we established a protocol to differentiate human-induced pluripotent stem cell-derived neural crest-like cells (iNCs) into PDLSC-like cells (iPDLSCs) using human PDL cell-derived extracellular matrix (ECM). However, it remained unclear what factors principally regulate the differentiation of iNCs into iPDLSCs. In this study, we aimed to identify the transcription factor regulating production of human PDL cell-derived ECM, which is responsible for the generation of iPDLSCs. We cultured iNCs on ECMs of two human PDL cell lines (HPDLC-3S and HPDLC-3U) and of human dermal fibroblasts (HDF). iNCs cultured on HPDLC-3U demonstrated higher iPDLSC-associated gene expression and mesenchymal differentiation capacity than cells cultured on HDF or HPDLC-3S. The transcription factor PAX9 was highly expressed in HPDLC-3U compared with HDF and HPDLC-3S. iNCs cultured on siPAX9-transfected HPDLC-3U displayed downregulation of iPDLSC-associated marker expression and adipocytic differentiation capacity relative to controls. Our findings suggest that PAX9 is one of the transcription factors regulating ECM production in human PDL cells, which is responsible for the differentiation of iNCs into iPDLSCs.

    DOI: 10.3390/biomedicines10102366

    Web of Science

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    PubMed

  • Effects of amplitude modulated discharge on growth of nanoparticles in TEOS/O<sub>2</sub>/Ar capacitively coupled plasma

    Kamataki, K; Nagamatsu, D; Yang, T; Abe, K; Yamamoto, A; Nagao, I; Arima, T; Otaka, M; Yamamoto, Y; Yamashita, D; Okumura, T; Yamashita, N; Itagaki, N; Koga, K; Shiratani, M

    AIP ADVANCES   12 ( 8 )   2022年8月   eISSN:2158-3226

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    出版者・発行元:Aip Advances  

    We investigate the effects of the amplitude modulation (AM) discharge method on the growth of nanoparticles and the relation between growth of nanoparticles and plasma generation in tetraethylorthosilicate (TEOS)/O2/Ar plasma. The laser-light scattering (LLS) intensity, which is proportional to the density and the sixth power of the size of nanoparticles in the Rayleigh scattering regime, decreases by 18% at an AM level of 10% and by 60% at an AM level of 50%. On the other hand, the ArI emission intensity, which is roughly proportional to plasma density, is higher than that for the continuous wave discharge. Thus, AM discharges suppress growth of nanoparticles in TEOS plasma. We have shown oscillations of the axial electric field Ez with the AM frequency for AM discharge by electric field measurement using an electro-optic probe. We have discussed that these fluctuations of Ez mainly lead to the vertical oscillation of the levitation position of nanoparticles trapped in the plasma sheath boundary region by taking into account the force balance equation in the axial direction on these negatively charged nanoparticles.

    DOI: 10.1063/5.0097691

    Web of Science

    Scopus

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