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間葉系幹細胞抽出物は歯根膜細胞の増殖を誘導する
Get PDF大阪歯科大学Osaka Dental University博士（歯学）Periodontitis is a chronic inflammatory disease caused primarily by gram-negative bacterial infection. The high incidence of this disease worldwide makes it a major oral health concern.1,2 The progression of periodontal disease leads to progressive loss of the attachment apparatus between the tooth and alveolar bone that supports the periodontal tissues, eventually leading to tooth extraction.3,4 Many therapies have been developed to regenerate periodontal tissues lost due to the disease. However, the extent of periodontal regeneration that can be achieved by these conventional regenerative therapies was limited, therefore the development of new periodontal regenerative therapies is highly desirable. The periodontal ligament is thin soft tissue that connects the teeth to alveolar bone and is responsible for physically buffering the occlusal forces applied on the teeth. In addition to its physical buffering function, the periodontal ligament has been shown to play an important role in wound healing and homeostasis of periodontal tissues by supplying progenitor/stem cells.5 It has also been demonstrated that periodontal tissue healing is dependent on the cell type that initially occupies the healing site, and that periodontal regeneration occurs only when periodontal ligament-derived cells proliferate and migrate to the wound area.5,6 The guided tissue regeneration (GTR) method has proved that periodontal regeneration occurs by inducing proliferation and migration of periodontal ligament cells to the root surface using a physical barrier membrane.7,8 This demonstrated that proliferation and migration of periodontal ligament cells are important phenomena for periodontal regeneration.
Mesenchymal stem cells (MSCs) are a stem cell population with multi-differentiation potential and are used for regenerative therapies for various diseases.9,10 In periodontal disease, transplantation of MSCs has been shown to regenerate periodontal tissues in many animal studies.11–14 However, the regenerative mechanism by MSCs remains largely unknown. Recently, it has been revealed that some of the therapeutic mechanisms of MSC depend on humoral factors released by MSC, and paracrine factors derived from MSCs have attracted attention as a novel cell-free regenerative material.15,16 We previously reported that periodontal tissues are regenerated by the transplantation of culture supernatants containing paracrine factors from MSCs.17 MSC paracrine factors are stored intracellularly before being released from the cells and may be recovered as cell extracts from cultured MSC. However, the function of MSC cell extracts is still largely unknown.
The purpose of this study was to investigate the effect of cell extracts from MSCs (MSC-extract) on the proliferation of periodontal ligament (PDL) cells, which play an important role in periodontal tissue regeneration.doctoral thesi
ポリカプロラクトン多孔質足場による骨形成効果に及ぼすEr: YAGパルスレーザー堆積法によるハイドロキシアパタイトコーティングの影響
Get PDF大阪歯科大学Osaka Dental University博士（歯学）Composite scaffolds obtained by the combination of biodegradable porous scaffolds and hydroxyapatite with bone regeneration potential are feasible materials for bone tissue engineering. However, most composite scaffolds have been fabricated by complicated procedures or under thermally harsh conditions. We have previously demonstrated that hydroxyapatite coating onto various substrates under a thermally mild condition was achieved by erbium-doped yttrium aluminum garnet (Er: YAG) pulsed laser deposition (PLD). The purpose of this study was to prepare a polycaprolactone (PCL) porous scaffold coated with the hydroxyapatite by the Er: YAG-PLD method. Hydroxyapatite coating by the Er: YAG-PLD method was confirmed by morphology, crystallographic analysis, and surface chemical characterization studies. When cultured on PCL porous scaffold coated with hydroxyapatite, rat bone marrow-derived mesenchymal stem cells adhered, spread, and proliferated well. The micro-CT and staining analyses after the implantation of scaffold into the critical-sized calvaria bone defect in rats indicate that PCL porous scaffold coated with hydroxyapatite demonstrates accelerated and widespread bone formation. In conclusion, PCL porous scaffold coated with hydroxyapatite obtained by the Er: YAG-PLD method is a promising material in bone tissue engineering.doctoral thesi
S-PRGフィラー溶出液はTNF-α刺激ヒト歯肉線維芽細胞のMMP-1分泌を抑制する
Get PDF大阪歯科大学Osaka Dental University博士（歯学）Tumor necrosis factor (TNF)-α is a major inflammatory factor that regulates tissue destruction in periodontitis and promotes the release of Matrix metalloproteinase (MMP) -1, which breaks down the gingival extracellular matrix. MMP-1 is one of the proteolytic enzymes that are capable of cleaving collagen and may play an important role in the destruction of connective tissue. The present study investigated the effects of Surface pre-reacted glass-ionomer (S-PRG) filler eluate on the secretion of MMP-1 by human gingival fibroblast (HGF) stimulated with the inflammatory cytokine TNF-α. S-PRG filler eluate did not alter Akt or nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65 phosphorylation levels in TNF-α-stimulated HGF. In the mitogen-activated protein kinase (MAPK) pathway, S-PRG filler eluate did not alter the phosphorylation level of p38 in TNF-α-stimulated HGF, but decreased that of c-Jun N-terminal kinase (JNK). These results will facilitate the development of new anti-inflammatory agents for periodontal disease using materials composed of S-PRG filler eluate.doctoral thesi
細胞内グルコース欠乏によるヒト歯肉組織の恒常性とオートファジーに及ぼす影響
Get PDF大阪歯科大学Osaka Dental University博士（歯学）Human gingival fibroblasts (HGnFs) maintain periodontal tissue homeostasis through active proliferation and migration. Clinically, it is considered that the wound-healing ability of the gingival tissue is maintained even in environments with insufficient supply of nutrients, such as glucose, immediately after periodontal surgery. However, the effects of such glucose-deficient environments on HGnFs remain unclear.This study aimed to investigate the effects of low-glucose environment on HGnFs homeostasis. We evaluated gingival wound healing by examining cell proliferation and migration and collagen synthesis in HGnFs cultured in 100, 50, 25, and 0 mg/dL glucose in vitro. The cellular stress levels were determined by measuring the lactate dehydrogenase (LDH) and reactive oxygen species (ROS) levels. The glucose metabolism of HGnFs in the low-glucose concentrations was studied by measuring glucose transporter type 1 (GLUT1) mRNA expression, glucose uptake assays, lactate and ATP productions. Molecular effects were examined with a focus on the LKB1-AMPK signaling pathway. Autophagy activity in glucose-deprived HGnFs was evaluated by measuring the levels of autophagy-related proteins.Low glucose levels increased cellular stress levels, autophagy activity, and enhanced glucose metabolism through the LKB1-AMPK signaling pathway, providing more ATPs to promote wound healing. Our results regarding glucose transfer suggest the rapid healing of gingival wounds.doctoral thesi
純チタン金属材料への大気圧窒素プラズマ処理が骨形成に与える影響
Get PDF大阪歯科大学Osaka Dental University博士（歯学）Pure titanium is used in dental implants owing to its excellent biocompatibility and physical properties. However, the aging of the material during storage is detrimental to the long-term stability of the implant after implantation. Therefore, in this study, we attempted to improve the surface properties and circumvent the negative effects of material aging on titanium implants by using a portable handheld nonthermal plasma device capable of piezoelectric direct discharge to treat pure titanium discs with nitrogen gas. We evaluated the osteogenic properties of the treated samples by surface morphology and elemental analyses, as well as in vitro and in vivo experiments. The results showed that nonthermal atmospheric-pressure nitrogen plasma can improve the hydrophilicity of pure titanium without damaging its surface morphology while introducing nitrogen-containing functional groups, thereby promoting cell attachment, proliferation, and osseointegration to some extent. Therefore, nitrogen plasma treatment may be a promising method for the rapid surface treatment of titanium implants.doctoral thesi
口腔扁平上皮癌細胞におけるdeferriferrichrysinの細胞死誘導効果
Get PDF大阪歯科大学Osaka Dental University博士（歯学）Certain siderophores have recently been reported to inhibit the growth of cancerous cells. Here, we assessed the cytotoxic effects of two siderophores, deferriferrichrysin (Dfcy) and bisucaberin, on the human oral squamous cell carcinoma (OSCC) cell lines HSC-3 and SAS. The iron-chelating ability of Dfcy and bisucaberin was measured using an iron assay kit utilizing the ferrozine chromogenic method. Cell viability was measured using the WST-8 assay, and caspase-3 activity was determined using a caspase-3 assay kit. Apoptotic, anti-apoptotic, and cell cycle markers were assessed by western blot analysis. Dfcy chelated ferric iron in a dose-dependent manner. In addition, 10 µg/mL bisucaberin chelated approximately 75% of the iron. Dfcy and bisucaberin inhibited the proliferation of HSC-3 and SAS cells in a dose-dependent manner. Bisucaberin-treated SAS cells exhibited significantly increased caspase-3 activation. Cleaved poly(ADP-ribose) polymerase (PARP) was also increased in bisucaberin-treated HSC-3 and SAS cells and in Dfcy-treated SAS cells. Bisucaberin treatments alone upregulated cleaved caspase-3 in SAS cells. However, Dfcy had no effect on OSCC cell apoptosis. The results show that bisucaberin exerts potential anticancer effects against SAS cells by inducing apoptosis.doctoral thesi
骨格性下顎前突症のトルコ鞍の形態に関する研究
Get PDF大阪歯科大学Osaka Dental University博士（歯学）We investigated the relationship between sella turcica size and maxillofacial morphol-ogy in skeletal Class III malocclusion among Japanese adults.The subjects were 115 patients visiting the Osaka Dental University Hospital between May 2014 and March 2019 with ∠ANB < 1.0º, negative overjet, and an absence of congenital abnormalities in the skeletal Class III malocclusion group. Individuals with 1.0º ≤ ∠ANB ≤ 4.0º were included as controls (n=122). Lateral cephalometric analysis was performed to measure the overbite depth indicator (ODI), the maximum anteroposterior width (W) of the sella turcica, depth (De) of the sella turcica, and diameter (Di) of the sella turcica; and inter-clinoid distance (I), among other parameters. In the skeletal Class III malocclusion group, Di and I were significantly smaller among the males, and De was significantly larger among females. In males, a positive correlation was observed between De and ODI. In females, a positive correlation was observed between W and Pog to NB, De and ∠ANB, ∠Occlusal pl to SN, and ∠GoGn to SN. These findings indicate the possibility of predicting maxillofacial growth based on sella turcica morphology, which could help improve orthodontic treatment.doctoral thesi
ブラジルバイーア州産プロポリスの口腔細菌叢へ与える影響について
Get PDF大阪歯科大学Osaka Dental University博士（歯学）Propolis is a resinous substance used by honey bees to construct hives. It is obtained by mixing pollen and plant resins with saliva and thus contains many natural substances. We previously reported that ethanol-extracted propolis (EEP) from Bahia, Brazil has antibacterial effects against the periodontopathic bacterium Porphyromonas gingivalis, although it may have very limited antibacterial effects against other oral bacterial species. In this study, we investigated the effect of EEP, which was provided by Yamada Bee Company, Inc. had on oral bacterial flora. Propolis treatment tended to decrease Fusobacterium, Prevotella, Peptostreptococcus, and Porphyromonas at the genus levels in oral bacterial flora. These results showed that EEP affects not only Porphyromonas spp., as shown in our previous in vitro study, but also many other bacterial species. We believe that propolis from Bahia, Brazil may be a potential reagent for oral health by regulating oral bacterial flora.doctoral thesi
健常成人における口腔筋機能療法のポッピング訓練による口腔機能改善に関する評価
Get PDF大阪歯科大学Osaka Dental University博士（口腔科学）【目的】口腔筋機能療法（以下，MFT）において舌を挙上させて舌背辺縁を口蓋周囲に密着させながら口を開け，ポンと音をたてるというポッピング訓練があるが，ポッピング訓練実施による舌圧の増加やそれに関連する口腔機能の改善についての有効性については明らかにされていない．本研究では，健常成人においてポッピング訓練と舌の口蓋への押し付け訓練による舌圧の増加等の口腔機能改善についての有効性を評価した．
【方法】健常成人24名をポッピング訓練群と舌の口蓋への押し付け訓練群に無作為に振り分けた．30回/1日，週5回×4週間の訓練を行った．口腔湿潤度，舌圧，咀嚼能力，オーラルディアドコキネシス（以下，ODK） /pa/，/ta/，/ka/について評価した．
【結果】舌圧，咀嚼能力，ODK /pa/，/ta/，/ka/で両群共に訓練前後で有意な差がみられた（p＜0.05）．訓練前後の変化量の比較では，両群に有意な差はみられなかった．
【結論】MFT訓練のひとつであるポッピング訓練は，舌の口蓋への押し付け訓練と同じく舌圧の増加のみならず，咀嚼能力およびODKの改善にも繋がることが示唆された．doctoral thesi
歯科衛生学生の社会人基礎力に対する臨床実習の効果 ならびに自己評価・他者評価に関する分析
Get PDF大阪歯科大学Osaka Dental University博士（口腔科学）先行研究において，臨床実習前後の歯科衛生学生の社会人基礎力は，3つの能力とも臨床実習後には上昇することを示唆する報告を行った．本研究では，経年的調査と並行して，自己評価の信憑性を検討するために，他者評価を取り入れて検討した．方法は，先行研究と同様の質問紙を用いて，歯科衛生士養成校3校（4年制大学，短期大学，専門学校）の臨床実習を履修する歯科衛生学生を対象に，質問紙調査を行った．うち1校の5か年分の実習前，実習中，実習後をより詳細に検討し，1か年分は，専任教員6名が自己評価と同様の手法で他者評価を行い，自己評価と他者評価の相違を検討した．その結果，いずれの歯科衛生士養成校においても，社会人基礎力の3つの能力は，実習前より実習後に有意に上昇した．3つの能力の中でも前に踏み出す力の平均値が，顕著な上昇を示した．12の能力要素について，実習前後での上昇率をスコア化して検討したところ，3校とも顕著に上昇する能力要素（働きかけ力，ストレスコントロール力，主体性）が，同じ傾向にあった．5か年分の検討では，実習前-実習中に有意な差を示す能力要素と，実習中-実習後に有意な差を示す能力要素があった．自己評価と他者評価の検討では，実習前はチームで働く力のみ，実習中と実習後は全ての能力に有意な差が認められた． 臨床実習は社会人基礎力の有機的かつ多面的な育成に有効である可能性が示唆された．doctoral thesi