Osaka Dental University Academic Repository / 大阪歯科大学学術リポジトリ
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脱分化脂肪細胞から作製した 3D スフェロイドの骨再生
Get PDF大阪歯科大学Osaka Dental University博士（歯学）Aim: Dedifferentiated fat (DFAT) cells, similar to mesenchymal stem cells (MSCs), are gaining attention owing to their minimally invasive nature. We aimed to evaluate the effects of 3D DFAT spheroids on bone regeneration by implantation into rat skull defects.
Methods: Adipose tissue was harvested from 8-week-old Sprague–Dawley rats, and DFAT was prepared using the ceiling culture method. DFAT was seeded onto threedimensional (3D) culture plates to produce spheroids, enhance cellular interactions, and mimic the in vivo environment. Bone defects were created on the skulls of 15-week-old rats using a 6.0-mm trephine burr. Collagen sponges were transplanted as controls, and 3D DFAT spheroid-seeded sponges were used in the experimental group.
Results: DFAT showed a fibroblast-like morphology. Flow cytometry revealed that DFAT expressed MSC markers but not hematopoietic stem cell markers. Bone analysis revealed increased bone mineral density, volume, and trabecular thickness in the experimental group. Hematoxylin–eosin staining in the control group revealed only a thin layer of fibrous tissue, whereas the experimental group showed new bone tissue
formation. The control group lacked Von Willebrand factor (vWf) expression, whereas the experimental group exhibited vWf expression, indicating the development of new blood vessels. Von Kossa staining revealed calcification only in the experimental group.
Conclusion: Spheroids alone are difficult to retain in bone defects, making their use in combination with collagen sponges more effective for bone regeneration. The study findings suggest that 3D DFAT spheroids may help regenerate rat skull defects, with potential implications for human bone regeneration.doctoral thesi
必須アミノ酸欠乏がヒト歯肉線維芽細胞に及ぼす生物学的影響
Get PDF大阪歯科大学Osaka Dental University博士（歯学）In this study, we aimed to investigate the effects of essential amino acid (EAA) deficiency on human gingival fibroblasts (HGFs). EAA deficiency inhibited HGF proliferation and migration, leading to impaired cellular functions. It also caused morphological changes, leading to thinner and less elongated cells, and suppressed collagen gene expression and synthesis. EAA deficiency promoted autophagy, as indicated by the increased expression levels of LC3 and decreased expression levels of p62. Overall, these findings suggest that EAA deficiency significantly impairs wound healing and tissue regeneration by HGFs. Therefore, adequate EAA intake is important to enhance tissue repair, particularly in periodontal treatment, by supporting the cellular functions necessary for healing.doctoral thesi
γ-D-グルタミル-メソ-ジアミノピメリン酸(iE-DAP) がヒト歯髄由来線維芽細胞のtissue inhibitor of metalloproteinase-1産生に及ぼす影響
Get PDF大阪歯科大学Osaka Dental University博士（歯学）The carious cavity contains a wide variety of oral microorganisms, which attempt to invade dental pulp tissue. Therefore, dental pulp tissue serves as the front line of biological defense through innate immune responses that recognize pathogen-derived molecules via pattern recognition receptors (PRRs). Nucleotide-binding oligomerization domain-containing protein (NOD)-1 is a type of PRR that detects γ-D-glutamyl-meso-diaminopimelic acid (iE-DAP) present in the peptidoglycan of gram-negative bacteria. Matrix metalloproteinases (MMPs) are important enzymes involved in physiological remodeling of connective tissue and in inflammatory tissue destruction. Tissue metalloproteinase inhibitors (TIMPs) are endogenous inhibitors of MMP-1 collagenase activity. This study aimed to investigate the effect of iE-DAP/NOD1 stimulation on TIMP-1 production and its signaling pathway in human dental pulp-derived fibroblasts (HDPFs). Our findings indicated that iE-DAP/NOD1 stimulation increased TIMP-1 production, reaching its peak at a concentration of 10 μg/mL. iE-DAP/NOD1 stimulation also enhanced the phosphorylation of ERK1/2 and JNK, both peaking at 5 min. U0126 (an ERK1/2 inhibitor) and SP600125 (a JNK inhibitor) suppressed TIMP-1 production. Overall, these results suggest that iE-DAP/NOD1 stimulation may enhance TIMP-1 production from HDPFs through a mechanism involving the phosphorylation of ERK1/2 and JNK.doctoral thesi
Mesenchymal stem cell-derived protein extract induces periodontal regeneration.
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NSC-3852 synergistically enhances the cytotoxicity of olaparib in oral squamous cell carcinoma.
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Effect of deterioration of swallowing functions on the frailty status in older adults: a longitudinal cohort study.
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家兎下顎骨におけるインプラント周囲骨欠損のクリティカルサイズについて
Get PDF大阪歯科大学Osaka Dental University博士（歯学）Background The mandible of the rabbit is considered a reliable model to be used to study bone regeneration in defects. The aim of the present study was to evaluate the formation of new bone around implants installed in defects of either 5 or 10 mm in the mandible of rabbits. Materials and methods In 12 rabbits, 3 mm deep circumferential defect, either 5 or 10 mm in diameter, were prepared bilaterally and an implant was placed in the center. A collagen membrane was placed to close the entrance. After 10 weeks, biopsies were taken, histological slides were prepared, and different regions of the defects were analyzed.
Results Similar amounts of new bone were found in both defects. However, most of the 5 mm defects were filled with new bone. New bone was observed closing the entrance of the defect and laid onto the implant surface. Only in a few cases the healing was incomplete. Despite a similar percentage of new bone found within the 10 mm defects, the healing was incomplete in most of the cases, presenting a low rate of bone formation onto the implant surface within the defect. Only one case presented the closure of the entrance. Conclusions The dimensions of the defect strongly influenced the healing so that a
circumferential marginal defect of 10 mm around an implant in the mandible body should be considered a critical-sized defect. The presence of the implant and of residues of teeth might have strongly influenced the healing.doctoral thesi
In situ ハイブリダイゼーション法を用いた歯原性嚢胞におけるペリオスチン発現パターンの解析
Get PDF大阪歯科大学Osaka Dental University博士（歯学）In situ hybridization (ISH) is an ultrasensitive technique for analyzing gene expression at the cellular level, enabling the detection of specific genes. It is an indispensable method, particularly in studies on developmental differentiation and morphogenesis, and is also attracting attention for its clinical application to biomarker detection. We focused on periostin (POSTN), an extracellular matrix protein, and examined the histological expression patterns of POSTN mRNA, particularly pathological splicing variants, in malignant tumors, such as breast cancer and tongue cancer, to investigate the potential of novel therapies targeting POSTN. However, POSTN mRNA staining has not yet been performed on the tissues of odontogenic tumors and cysts. Therefore, we herein examined the expression patterns of POSTN mRNA in human explants of dentigerous cysts, orthokeratinized odontogenic cysts, and odontogenic keratocysts, which are commonly encountered in clinical practice, using ISH. The results obtained suggest that POSTN mRNA was mainly expressed in fibroblasts in all odontogenic cysts, and the intensity of expression appeared to vary depending on the presence of inflammation. In odontogenic keratocysts after decompression, POSTN mRNA was highly expressed in the basal cell layer. ISH enabled the cellular localization of POSTN to be identified, which was difficult by immunohistochemistry, showing the potential of its clinical application to biomarker detection not only in odontogenic cysts, but also in all diseases of the oral and maxillofacial region.doctoral thesi
β1インテグリン/FAKシグナル伝達経路は、ヒト歯肉上皮細胞株 Ca9-22細胞におけるインターロイキン-8の産生を調節する
Get PDF大阪歯科大学Osaka Dental University博士（歯学）Interleukin-8 (IL-8), one of the proinflammatory factors in human tissues, plays an important role in inflammation. Type IV collagen, a key component of the basement membrane, interacts with integrins, which are primary receptors of the extracellular matrix (ECM). Integrins are essential for regulating various cellular behaviors and signal transduction pathways. The relationship between type IV collagen, β1 integrin, and gingival epithelial cells is poorly understood. The aim of the present study was to elucidate the effect of interaction between type IV collagen and β1 integrin on IL-8 secretion in human gingival epithelial cells (Ca9-22).doctoral thesi