Journal of Oral Biosciences最新文献_第4页

RANKL enhances the expression of PEPT1/SLC15A1 and PEPT2/SLC15A2 in RAW264.7 cells RANKL可增强RAW264.7细胞中PEPT1/SLC15A1和PEPT2/SLC15A2的表达

IF 2.3 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Journal of Oral Biosciences

Pub Date : 2025-12-20 DOI: 10.1016/j.job.2025.100724

Mana Shintani , Wakana Sugimoto , Hiroshi Inoue , Nagako Sougawa , Seiji Goda , Aki Nishiura

Objectives

Muramyl dipeptide (MDP), a bacterial cell wall component, is recognized by NOD2 and vital in innate immune responses, including inflammatory cytokine production. MDP is transported into the cells via PEPT1/SLC15A1 and PEPT2/SLC15A2, which are members of the proton-coupled oligopeptide transporter family within the SLC15 solute carrier group. Although the effects of RANKL stimulation on certain transporters are known, its effects on the SLC15 family remain unclear. This study aimed to clarify the effects of RANKL stimulation on PEPT1/SLC15A1 and PEPT2/SLC15A2 in RAW264.7 cells and determine their role in osteoclast differentiation.

Methods

RAW264.7 cells were stimulated with RANKL and MDP. Expression levels of NOD2, PEPT1/SLC15A1, PEPT2/SLC15A2, cathepsin K, and NFATc1 were analyzed via Western blotting. Osteoclast differentiation was evaluated using a tartrate-resistant acid phosphatase (TRAP) activity assay.

Results

RANKL stimulation increased NOD2, PEPT1/SLC15A1, and PEPT2/SLC15A2 expression in RAW264.7 cells. Colistin and polymyxin B, which are PEPT1 and PEPT2 inhibitors, respectively, did not affect the stimulation of cells with RANKL alone. However, RANKL and MDP co-stimulation suppressed the RANKL/MDP-induced increase in TRAP activity and cathepsin K and NFATc1 expression.

Conclusions

RANKL stimulation increased PEPT1/SLC15A1 and PEPT2/SLC15A2 levels in RAW264.7 cells, suggesting an increase in the intracellular uptake of MDP. This may promote osteoclast differentiation, potentially through NOD2activation or NOD2-independent mechanisms.

目的：muramyl二肽（MDP）是一种细菌细胞壁成分，被NOD2识别，在先天性免疫反应中至关重要，包括炎症细胞因子的产生。MDP通过PEPT1/SLC15A1和PEPT2/SLC15A2转运到细胞中，它们是SLC15溶质载体群中质子偶联寡肽转运蛋白家族的成员。虽然RANKL刺激对某些转运蛋白的影响是已知的，但其对SLC15家族的影响尚不清楚。本研究旨在阐明RANKL刺激对RAW264.7细胞PEPT1/SLC15A1和PEPT2/SLC15A2的影响，并确定其在破骨细胞分化中的作用。Western blotting分析NOD2、PEPT1/SLC15A1、PEPT2/SLC15A2、cathepsin K、NFATc1的表达水平。使用抗酒石酸酸性磷酸酶（TRAP）活性测定来评估破骨细胞分化。结果rankl刺激可增加RAW264.7细胞中NOD2、PEPT1/SLC15A1和PEPT2/SLC15A2的表达。粘菌素和多粘菌素B分别是PEPT1和PEPT2抑制剂，单独使用RANKL对细胞的刺激没有影响。然而，RANKL和MDP共同刺激抑制了RANKL/MDP诱导的TRAP活性和组织蛋白酶K和NFATc1表达的增加。结论rankl刺激增加了RAW264.7细胞中PEPT1/SLC15A1和PEPT2/SLC15A2的水平，表明MDP的细胞内摄取增加。这可能通过nod2激活或nod2独立机制促进破骨细胞分化。

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引用次数: 0

Odontogenic ameloblast-associated protein (ODAM)-enriched coating improves adhesion of junctional epithelial cells to composite resin via AKT activation 富牙源性成釉细胞相关蛋白（ODAM）涂层通过激活AKT改善结上皮细胞对复合树脂的粘附

IF 2.3 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Journal of Oral Biosciences

Pub Date : 2025-12-13 DOI: 10.1016/j.job.2025.100725

Masayoshi Takaman , Keishi Otsu , Shojiro Ikezaki , Mamoru Noda , Hidemitsu Harada

Objectives

Although the junctional epithelium (JE) forms a biological seal around the teeth, attachment is often unstable in cases of subgingival extension of the composite resin (CR) margins. Odontogenic ameloblast-associated protein (ODAM), a JE-specific extracellular matrix protein, regulates adhesion and cytoskeletal dynamics. This study investigated whether an ODAM-enriched coating on CR surfaces enhances JE cell function, and elucidated the underlying molecular mechanisms.

Methods

An ODAM-enriched fraction was purified and coated onto CR discs. The mHAT-JE01 cells were cultured on either uncoated or ODAM-coated surfaces. Cell adhesion and proliferation were assessed by fluorescence imaging and Ki67 staining, cell migration by wound healing, and cell morphology by scanning electron microscopy. The expression of focal adhesion–related molecules, actin filaments, and phosphorylated AKT was analyzed using immunofluorescence and quantitative polymerase chain reaction. An AKT inhibitor was used to evaluate the need for AKT signaling.

Results

ODAM coating of CR surfaces significantly enhanced the adhesion, proliferation, and migration of JE cells, reinforcing actin stress fibers, promoting vinculin accumulation at the cell periphery and protrusions, and increasing phosphorylated AKT levels. ODAM coating induced its own expression, suggesting a novel positive feedback mechanism that amplifies the biological effects. AKT inhibition suppressed ODAM-mediated changes, confirming that AKT is the central hub for focal adhesion–related signaling and epithelial responses.

Conclusions

These findings support the role of ODAM as a bioactive molecule that contributes to subgingival margin stabilization, mitigates peri-restorative inflammation, and enhances long-term periodontal and restorative outcomes.

目的：虽然结合上皮（JE）在牙齿周围形成生物密封，但复合树脂（CR）边缘龈下延伸时附着往往不稳定。牙源性成釉细胞相关蛋白（ODAM）是一种je特异性的细胞外基质蛋白，可调节粘附和细胞骨架动力学。本研究探讨了CR表面的odam富集涂层是否能增强乙脑细胞功能，并阐明了其潜在的分子机制。方法纯化富含san odam的组分，包被于CR光盘上。mHAT-JE01细胞分别在未包被或odam包被的表面上培养。通过荧光成像和Ki67染色评估细胞粘附和增殖，通过伤口愈合评估细胞迁移，通过扫描电镜观察细胞形态。利用免疫荧光和定量聚合酶链反应分析局灶黏附相关分子、肌动蛋白丝和磷酸化AKT的表达。AKT抑制剂用于评估AKT信号的必要性。结果sodam包覆CR表面可显著增强乙脑细胞的粘附、增殖和迁移能力，增强肌动蛋白应激纤维，促进细胞外周和突起的血管素积累，提高磷酸化AKT水平。ODAM涂层诱导自身表达，提示一种新的正反馈机制，放大了生物效应。AKT的抑制抑制了odam介导的变化，证实了AKT是局灶性粘连相关信号传导和上皮反应的中枢。这些发现支持ODAM作为一种生物活性分子的作用，有助于牙龈下边缘稳定，减轻修复期炎症，并提高长期牙周和修复效果。

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引用次数: 0

Dipeptidyl peptidase from Streptococcus anginosus with substrate specificity for Xaa-Pro/Ala and potential impact on tumor immunity 具有Xaa-Pro/Ala底物特异性的血管链球菌二肽基肽酶及其对肿瘤免疫的潜在影响

IF 2.3 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Journal of Oral Biosciences

Pub Date : 2025-12-11 DOI: 10.1016/j.job.2025.100723

Shu Suzuki , Toshitaka Miura , Yu Shimoyama , Yuko Ohara-Nemoto , Takayuki K. Nemoto , Hiroyuki Yamada , Taichi Ishikawa

Objectives

Streptococcus anginosus, an oral commensal bacterium, is a potential risk factor for malignancies of the oral cavity and upper gastrointestinal tract. Although this species harbors an Xaa-Pro dipeptidyl peptidase (DPP), its enzymatic characteristics and potential role in tumor-associated immune modulation remain unclear. In this study, S. anginosus Xaa-Pro DPP was characterized and its ability to cleave chemokine-related peptides was evaluated.

Methods

Six oral streptococcal strains were analyzed for DPP activity using fluorogenic dipeptidyl substrates. The gene encoding S. anginosus Xaa-Pro DPP was cloned and the recombinant enzyme was purified and characterized. The enzymatic activity of the peptidase against synthetic dipeptidyl 7-amino-4-methylcoumarin (MCA) and chemokine-derived peptides was assessed using fluorescence assays and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The effect of the DPP4 inhibitor, P32/98, was also examined.

Results

S. anginosus had the highest DPP activity among the tested oral streptococci, particularly towards Gly-Pro- and Lys-Ala-MCA. The recombinant enzyme selectively removed Xaa-Pro and Xaa-Ala dipeptides and the N-terminal Lys-Pro dipeptide from the CXCL12-derived peptide, whereas incretins were minimally affected. The enzymatic activity of Xaa-Pro DPP against synthetic substrates was markedly inhibited by P32/98.

Conclusion

S. anginosus Xaa-Pro DPP has specificity for N-terminal Xaa-Pro/Ala in peptides and exhibits the ability to inactivate chemokine-related peptides, suggesting a potential contribution to immune dysregulation in the tumor microenvironment.

目的：血管链球菌是一种口腔共生菌，是口腔和上消化道恶性肿瘤的潜在危险因素。尽管该物种含有Xaa-Pro二肽基肽酶（DPP），但其酶学特性及其在肿瘤相关免疫调节中的潜在作用尚不清楚。本研究对S. anginosus Xaa-Pro DPP进行了表征，并对其切割趋化因子相关肽的能力进行了评价。方法采用荧光二肽基底物对6株口服链球菌进行DPP活性分析。克隆了血管棘鱼Xaa-Pro DPP基因，并对重组酶进行了纯化和鉴定。利用荧光分析和基质辅助激光解吸/电离飞行时间质谱法评估了肽酶对合成二肽基7-氨基-4-甲基香豆素（MCA）和趋化因子衍生肽的酶活性。对DPP4抑制剂P32/98的作用也进行了检测。在测试的口服链球菌中，血管球菌具有最高的DPP活性，特别是对Gly-Pro-和Lys-Ala-MCA。重组酶选择性地从cxcl12衍生的肽中去除Xaa-Pro和Xaa-Ala二肽以及n端Lys-Pro二肽，而肠促胰岛素受到的影响最小。p32 /98明显抑制Xaa-Pro DPP对合成底物的酶活性。anginosus Xaa-Pro DPP对肽中的n端Xaa-Pro/Ala具有特异性，并表现出灭活趋化因子相关肽的能力，这表明在肿瘤微环境中可能有助于免疫失调。

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引用次数: 0

Choline acetyltransferase as a marker for Merkel cells in mucosal and skin tissues 胆碱乙酰转移酶作为粘膜和皮肤组织中默克尔细胞的标记物

IF 2.3 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Journal of Oral Biosciences

Pub Date : 2025-12-09 DOI: 10.1016/j.job.2025.100722

Ranhui Xi , Jiaxin Liu , Shahd Almasswary , Xin Xu , Marco Tizzano

Objectives

To investigate the expression of choline acetyltransferase (ChAT), the enzyme responsible for acetylcholine (ACh) synthesis, in Merkel cells in hairy and glabrous skin, including whiskers, palms, and oral mucosa, and to evaluate whether ChAT is a general molecular marker for Merkel cells that could indicate a potential cholinergic role in mechanosensory signaling.

Methods

Immunofluorescence staining was performed on skin and oral epithelial tissues of ChAT-tau-enhanced green fluorescent protein (ChAT-eGFP) transgenic mice. Co-expression of ChAT-eGFP and the Merkel cell marker, keratin 8 (KRT8), was analyzed to identify ChAT⁺ Merkel cells. Their spatial organization and association with nerve endings were examined, and age-related changes in the Merkel cell count in the palate were quantified.

Results

Merkel cells consistently co-expressed ChAT-eGFP and KRT8 in skin and mucosal tissues, and they were organized into characteristic clusters, closely associated with nerve fibers. There was also ChAT expression in taste-related structures and cholinergic epithelial cells. In aged mice, Merkel cell count in the palate was significantly reduced, particularly in anterior rugae.

Conclusions

This study identified ChAT as a novel and general marker for Merkel cells in skin and oral tissues, revealing a previously unrecognized cholinergic component of Merkel cell signaling. These findings suggest a potential role of ACh in mechanotransduction and tactile sensation. By establishing ChAT as a Merkel cell marker, this study provides new insights into somatosensory biology and provides a molecular framework for exploring cholinergic mechanisms in tactile perception. These findings may inform therapeutic strategies for sensory disorders of skin and oral mucosa.

目的研究胆碱乙酰转移酶（ChAT）在毛性和无毛性皮肤（包括胡须、手掌和口腔粘膜）Merkel细胞中的表达，并评估ChAT是否为Merkel细胞的一般分子标记物，可能表明胆碱能在机械感觉信号传导中发挥潜在的作用。方法对chat -tau增强绿色荧光蛋白（ChAT-eGFP）转基因小鼠皮肤和口腔上皮组织进行免疫荧光染色。通过分析ChAT- egfp与默克尔细胞标志物角蛋白8 （keratin 8, KRT8）的共表达来鉴定ChAT+默克尔细胞。研究了它们的空间组织和与神经末梢的联系，并量化了上颚默克尔细胞计数的年龄相关变化。结果erkel细胞在皮肤和粘膜组织中一致共表达ChAT-eGFP和KRT8，并呈特征性簇状排列，与神经纤维密切相关。在味觉相关结构和胆碱能上皮细胞中也有ChAT的表达。在老年小鼠中，上颚的默克尔细胞计数明显减少，尤其是在前颊。本研究发现ChAT是皮肤和口腔组织中默克尔细胞的一种新的通用标记物，揭示了默克尔细胞信号传导中以前未被识别的胆碱能成分。这些发现提示乙酰胆碱在机械传导和触觉感觉中的潜在作用。通过建立ChAT作为Merkel细胞标志物，本研究为躯体感觉生物学提供了新的见解，并为探索触觉感知中的胆碱能机制提供了分子框架。这些发现可能为皮肤和口腔黏膜感觉障碍的治疗策略提供信息。

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引用次数: 0

New insights into the influence of surface roughness of titanium and zirconia-based materials in the regenerative behavior of human gingival keratinocytes 钛和氧化锆基材料表面粗糙度对人牙龈角质形成细胞再生行为影响的新见解

IF 2.3 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Journal of Oral Biosciences

Pub Date : 2025-12-08 DOI: 10.1016/j.job.2025.100712

Irati Fernandez-de-Mendiola , Igor Irastorza , Sara Martin-Iglesias , Gaskon Ibarretxe , Fernando-José Unda , Raphael S. Wagner , Unai Silvan , Lucía Jiménez-Rojo

Objectives

This study aimed to investigate the impact of implant composition and surface modifications on the behavior of human gingival keratinocytes, a natural source of peri-implant epithelium.

Methods

The materials used in this study were titanium grade 4 and 3 mol% yttria-stabilized tetragonal zirconia polycrystal (3Y-TZP) discs with various surface modifications (e.g., polished, machined, sandblasted, and/or acid-etched). Surface roughness and wettability were first characterized, followed by culturing human gingival keratinocytes on these substrates. Cell morphology, adhesion, extracellular matrix (ECM) synthesis, cell cycle progression, and differentiation were analyzed using immunofluorescence and quantitative polymerase chain reaction (qPCR).

Results

The chemical composition and surface topography of the materials markedly influenced keratinocyte behavior. Specifically, reduced surface roughness enhanced the formation of mature focal adhesions, increased laminin-332 deposition, and promoted keratinocyte motility and proliferation. After 24 h of culture, smooth surfaces showed higher cell densities and a greater proportion of cycling cells. Interestingly, after one week of culture, smooth ceramic materials presented significantly higher gingival keratinocyte densities than smooth titanium surfaces. Furthermore, zirconia-based ceramic surfaces induced the expression of junctional epithelial markers, including keratin 19 (KRT19) and odontogenic ameloblast-associated protein (ODAM), while maintaining keratin 14 (KRT14) expression.

Conclusions

Our results demonstrate that biomaterials with smooth surfaces, especially of zirconia, induce pro-regenerative biological responses conducive to the development of a functional peri-implant epithelium.

目的研究种植体成分和表面修饰对人牙龈角化细胞（种植体周围上皮的天然来源）行为的影响。方法本研究中使用的材料是钛级4和3mol %钇稳定的四方氧化锆多晶（3Y-TZP）圆盘，具有各种表面修饰（如抛光，机械加工，喷砂和/或酸蚀）。首先表征表面粗糙度和润湿性，然后在这些基质上培养人牙龈角质形成细胞。利用免疫荧光和定量聚合酶链反应（qPCR）分析细胞形态、粘附、细胞外基质（ECM）合成、细胞周期进展和分化。结果材料的化学成分和表面形貌对角质形成细胞的行为有显著影响。具体来说，表面粗糙度的降低促进了成熟局灶粘连的形成，增加了层粘连蛋白332的沉积，促进了角化细胞的运动和增殖。培养24 h后，光滑表面细胞密度更高，循环细胞比例更大。有趣的是，培养一周后，光滑陶瓷材料的牙龈角质细胞密度明显高于光滑钛表面。此外，氧化锆陶瓷表面诱导了接合上皮标志物的表达，包括角蛋白19 （KRT19）和成牙釉质相关蛋白（ODAM），同时维持了角蛋白14 （KRT14）的表达。结论表面光滑的生物材料，尤其是氧化锆材料，可诱导促再生的生物学反应，有利于种植体周围上皮的发育。

{"title":"New insights into the influence of surface roughness of titanium and zirconia-based materials in the regenerative behavior of human gingival keratinocytes","authors":"Irati Fernandez-de-Mendiola , Igor Irastorza , Sara Martin-Iglesias , Gaskon Ibarretxe , Fernando-José Unda , Raphael S. Wagner , Unai Silvan , Lucía Jiménez-Rojo","doi":"10.1016/j.job.2025.100712","DOIUrl":"10.1016/j.job.2025.100712","url":null,"abstract":"<div><h3>Objectives</h3><div>This study aimed to investigate the impact of implant composition and surface modifications on the behavior of human gingival keratinocytes, a natural source of peri-implant epithelium.</div></div><div><h3>Methods</h3><div>The materials used in this study were titanium grade 4 and 3 mol% yttria-stabilized tetragonal zirconia polycrystal (3Y-TZP) discs with various surface modifications (e.g., polished, machined, sandblasted, and/or acid-etched). Surface roughness and wettability were first characterized, followed by culturing human gingival keratinocytes on these substrates. Cell morphology, adhesion, extracellular matrix (ECM) synthesis, cell cycle progression, and differentiation were analyzed using immunofluorescence and quantitative polymerase chain reaction (qPCR).</div></div><div><h3>Results</h3><div>The chemical composition and surface topography of the materials markedly influenced keratinocyte behavior. Specifically, reduced surface roughness enhanced the formation of mature focal adhesions, increased laminin-332 deposition, and promoted keratinocyte motility and proliferation. After 24 h of culture, smooth surfaces showed higher cell densities and a greater proportion of cycling cells. Interestingly, after one week of culture, smooth ceramic materials presented significantly higher gingival keratinocyte densities than smooth titanium surfaces. Furthermore, zirconia-based ceramic surfaces induced the expression of junctional epithelial markers, including keratin 19 (<em>KRT19</em>) and odontogenic ameloblast-associated protein (<em>ODAM</em>), while maintaining keratin 14 (<em>KRT14</em>) expression.</div></div><div><h3>Conclusions</h3><div>Our results demonstrate that biomaterials with smooth surfaces, especially of zirconia, induce pro-regenerative biological responses conducive to the development of a functional peri-implant epithelium.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"68 1","pages":"Article 100712"},"PeriodicalIF":2.3,"publicationDate":"2025-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145747945","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Solute carrier family 9 member A5 regulated by TGF-β is necessary for dental enamel formation 由TGF-β调控的溶质载体家族9成员A5是牙釉质形成所必需的

IF 2.3 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Journal of Oral Biosciences

Pub Date : 2025-12-04 DOI: 10.1016/j.job.2025.100719

Hayato Takano , Risako Chiba-Ohkuma , Saeko Kobayashi , Ryuji Yamamoto , Rei Kataoka , Yuri Miyakawa , Yoshinobu Asada , Yasuo Yamakoshi

Objectives

In this study, we aimed to elucidate the functional significance of transforming growth factor-β (TGF-β)-regulated SLC9A5 (NHE5) in enamel formation.

Methods

Mouse ameloblast-derived mHAT9d cells were treated with TGF-β1, TGF-β2, or TGF-β3, and the expression of Slc9a family members was analyzed via RNA sequencing (RNA-seq) and quantitative PCR (qPCR). Slc9a5-deficient mice were generated to examine enamel morphology. Enamel volume, mineral density, and structure were assessed using micro-computed tomography (μCT), scanning electron microscopy (SEM), and electron probe microanalysis (EPMA). Protein expression was analyzed via SDS–PAGE, western blotting, and immunohistochemistry.

Results

RNA-seq and qPCR analyses revealed that TGF-β1 and TGF-β3 strongly induced Slc9a5 in a dose-dependent manner; however, TGF-β2 had a minimal effect. In Slc9a5-deficient mice, incomplete crown formation and pit-like defects were evident on postnatal day 5; however, enamel protein profiles and thickness formation at day 11 were comparable to those in wild-type mice. In contrast, by day 70, μCT revealed marked thinning of enamel and reduced mineral density, SEM showed cracks and surface defects, and EPMA demonstrated significantly lower calcium-to-phosphorus (Ca/P) molar ratios than in wild-type mice. These findings indicate that loss of Slc9a5 slightly affects protein secretion but causes defective mineral maturation and enamel fragility.

Conclusions

SLC9A5 is a downstream target of TGF-β signaling, which is indispensable for enamel maturation because it maintains ion transport and extracellular pH homeostasis in ameloblasts. Its deficiency leads to reduced mineral density, altered Ca/P composition, and progressive enamel loss. These findings underscore the essential role of SLC9A5 in the long-term structural stability of enamel.

目的探讨转化生长因子-β (TGF-β)调控的SLC9A5 （NHE5）在牙釉质形成中的作用。方法用TGF-β1、TGF-β2、TGF-β3处理小鼠成釉细胞mHAT9d细胞，通过RNA测序（RNA-seq）和定量PCR （qPCR）分析Slc9a家族成员的表达。制备slc9a5缺陷小鼠，观察其牙釉质形态。采用显微计算机断层扫描（μCT）、扫描电镜（SEM）和电子探针显微分析（EPMA）评估牙釉质体积、矿物质密度和结构。通过SDS-PAGE、western blotting和免疫组织化学分析蛋白表达。结果rna -seq和qPCR分析显示，TGF-β1和TGF-β3对Slc9a5的诱导作用呈剂量依赖性；而TGF-β2的作用微乎其微。slc9a5缺陷小鼠在出生后第5天出现不完全冠形成和坑状缺陷；然而，第11天的牙釉质蛋白谱和厚度形成与野生型小鼠相当。70 d时，μCT显示牙釉质明显变薄，矿物质密度降低，SEM显示裂纹和表面缺陷，EPMA显示钙磷摩尔比明显低于野生型小鼠。这些结果表明，Slc9a5缺失会轻微影响蛋白质分泌，但会导致矿物质成熟缺陷和牙釉质脆性。结论sslc9a5是TGF-β信号转导的下游靶点，在成釉细胞中维持离子转运和细胞外pH稳态，对釉质成熟至关重要。它的缺乏导致矿物质密度降低，钙磷组成改变，并导致牙釉质逐渐丧失。这些发现强调了SLC9A5在牙釉质长期结构稳定性中的重要作用。

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引用次数: 0

Effects of Porphyromonas gingivalis-derived lipopolysaccharide on the barrier function of cultured junctional epithelial cells 牙龈卟啉单胞菌来源的脂多糖对培养的结膜上皮细胞屏障功能的影响

IF 2.3 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Journal of Oral Biosciences

Pub Date : 2025-12-02 DOI: 10.1016/j.job.2025.100721

Ryo Aizawa , Masaru Sunaga , Keisuke Tanaka , Marika Sugano , Daisuke Saito , Koki Okada , Junichi Tanaka , Kenji Mishima , Matsuo Yamamoto

Objectives

The junctional epithelium (JE) plays an important role in maintaining the protective integrity of periodontal tissues by forming an epithelial barrier that impedes bacterial invasion. This study examined the effects of Porphyromonas gingivalis lipopolysaccharide (PG-LPS) on JE-1 cells, a mouse-derived junctional epithelial cell line.

Methods

JE-1 cells were exposed to PG-LPS, and the effects on cell viability, migration, gene expression, protein expression, and barrier function at various time points were evaluated according to the experimental type.

Results

PG-LPS (1 μg/mL) did not affect JE-1 cell viability but significantly inhibited migration. Treatment with PG-LPS upregulated inflammatory genes (Il-6, Tlr2, Tlr4, Traf6) and altered the expression of chemokines (increased Cxcl2 and decreased Cxcl10) and protective factors (increased Nfe2l2 and Slpi). Notably, 1 μg/mL of PG-LPS increased permeability and decreased adhesion molecule expression (Cdh1 and Itgb4), whereas 10 μg/mL showed a non-linear response (apparent permeability [cm/s]: 0.868, 0.915, 1.416, 1.728, 1.224, and 1.176 with 0, 0.1, 0.5, 1, 5, and 10 μg/mL respectively), indicating compensatory mechanisms.

Conclusions

PG-LPS disrupts JE barrier function and alters its immunomodulatory properties, potentially contributing to periodontitis progression. These findings enhance our understanding of the underlying causes of periodontitis and inform targeted treatments to maintain JE defense against periodontopathogenic bacteria.

目的结膜上皮（JE）在维持牙周组织的保护性完整性方面起着重要作用，它通过形成一个上皮屏障来阻止细菌的入侵。本研究检测了牙龈卟啉单胞菌脂多糖（PG-LPS）对小鼠来源的连接上皮细胞系JE-1细胞的影响。方法采用PG-LPS处理sbe -1细胞，根据不同的实验类型，观察不同时间点对sbe -1细胞活力、迁移、基因表达、蛋白表达和屏障功能的影响。结果spg - lps （1 μg/mL）对JE-1细胞活力无明显影响，但明显抑制其迁移。PG-LPS可上调炎症基因（Il-6、Tlr2、Tlr4、Traf6），改变趋化因子（Cxcl2升高、Cxcl10降低）和保护因子（Nfe2l2和Slpi升高）的表达。其中，1 μg/mL的PG-LPS增加了通透性，降低了黏附分子（Cdh1和Itgb4）的表达，而10 μg/mL的PG-LPS在0、0.1、0.5、1、5和10 μg/mL的作用下表现为非线性响应（表观通透性[cm/s]分别为0.868、0.915、1.416、1.728、1.224和1.176），提示存在补偿机制。结论spg - lps破坏乙脑屏障功能，改变其免疫调节特性，可能促进牙周炎的进展。这些发现增强了我们对牙周炎的潜在原因的理解，并为有针对性的治疗提供了信息，以维持乙脑对牙周致病菌的防御。

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引用次数: 0

Dynamin 2 is involved in osteoblast migration by regulating the organization of F-actin 动力蛋白2通过调节f -肌动蛋白的组织参与成骨细胞的迁移

IF 2.3 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Journal of Oral Biosciences

Pub Date : 2025-12-02 DOI: 10.1016/j.job.2025.100720

Takumi Moriya , A. Surong , Nanami Tatsumi , Hiroshi Yamada , Fumiko Takemoto , Hiroshi Kamioka , Hirohiko Okamura , Mika Ikegame

Objectives

Dynamin, a GTPase that regulates membrane dynamics, has recently been implicated in actin cytoskeletal remodeling. This study aimed to elucidate the role of dynamin in osteoblast migration by examining the effects of dynamin inhibition on the localization and organization of F-actin and dynamin 2 in MC3T3-E1 cells.

Methods

MC3T3-E1 cells were treated with dynamin inhibitors (Dyngo 4a and Dynole 34-2), and cell migration was assessed using a wound-healing assay. Fluorescent staining was performed to analyze the intracellular localization of F-actin and dynamin 2.

Results

Dynamin inhibition significantly reduced the migration of MC3T3-E1 cells. Fluorescence analysis revealed a marked decrease in the accumulation and colocalization of F-actin and dynamin 2 at the protrusion edge. Additionally, dynamin inhibition suppressed the formation of lamellipodia and stress fibers while promoting the appearance of abnormal F-actin clusters in the cytoplasm.

Conclusions

These findings suggest that dynamin plays an essential role in osteoblast migration by regulating actin cytoskeletal remodeling, particularly through the formation of lamellipodia and stress fibers.

目的动态蛋白是一种调节膜动力学的GTPase，最近被认为与肌动蛋白细胞骨架重塑有关。本研究旨在通过检测动力蛋白抑制对MC3T3-E1细胞F-actin和动力蛋白2的定位和组织的影响，阐明动力蛋白在成骨细胞迁移中的作用。方法用动力蛋白抑制剂（dyngo4a和Dynole 34-2）处理smc3t3 - e1细胞，采用伤口愈合实验评估细胞迁移。荧光染色分析F-actin和dynamin 2在细胞内的定位。结果dynamin抑制显著降低MC3T3-E1细胞的迁移能力。荧光分析显示，F-actin和dynamin 2在突起边缘的聚集和共定位明显减少。此外，动力蛋白抑制抑制板足和应激纤维的形成，同时促进细胞质中异常f -肌动蛋白簇的出现。结论动力蛋白通过调节肌动蛋白细胞骨架重塑，特别是通过板足和应力纤维的形成，在成骨细胞迁移中起重要作用。

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引用次数: 0

Evaluation of Mycobacterium-derived plasmids for application in oral Actinomyces species 分枝杆菌源质粒在口腔放线菌中的应用评价

IF 2.3 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Journal of Oral Biosciences

Pub Date : 2025-12-02 DOI: 10.1016/j.job.2025.100718

Sakiko Ohara , Yijuan Sheng , Yuki Nishiya , Ikue Tosa , Katsuki Takebe , Yuki Arimura , Hiroshi Mese , Naoko Ohara , Naoya Ohara

Objectives

Genetic manipulation tools are essential for elucidating the pathogenic mechanisms of microorganisms. Several species of Actinomyces, including A. israelii, are present in the oral cavity and they are the causative agents of actinomycosis. However, efficient gene-editing tools for these species have not yet been developed. In this study, the aim was to evaluate the introduction of foreign genes into Actinomyces using plasmids derived from Mycobacterium, which belong to the same class as Actinomycetes.

Methods

A truncated derivative of pYT923, pYT923S, which contains the replication origin of the M. scrofulaceum plasmid pMSC262 was constructed and introduced into A. israelii by electrotransformation.

Results

pYT923S was successfully introduced into A. israelii. The transformation efficiency of A. israelii was approximately 7–66 CFU/μg of DNA, and all transformed colonies harbored pYT923S. The plasmid recovered from A. israelii replicated in Escherichia coli.

Conclusions

pYT923S was introduced into and maintained within A. israelii. Therefore, the pYT923S vector represents a useful genetic tool for Actinomyces and it is expected to facilitate future studies on the biology and pathogenicity of Actinomyces.

目的基因操作工具是阐明微生物致病机制的必要工具。包括以色列芽孢杆菌在内的几种放线菌存在于口腔中，它们是放线菌病的病原体。然而，针对这些物种的有效基因编辑工具尚未开发出来。在本研究中，目的是评估外源基因引入放线菌的质粒来自分枝杆菌，属于同一类放线菌。方法构建pYT923的截断衍生物pYT923S，其中pYT923S含有M. scfulaceum质粒pMSC262的复制起点，并通过电转化将其导入以色列拟南芥。结果成功将spyt923s引入以色列拟南芥。以色列拟南芥的转化效率约为7 ~ 66 CFU/μg DNA，所有转化菌落均携带pYT923S。从以色列芽孢杆菌中回收的质粒在大肠杆菌中复制。结论spyt923s在以色列拟南蝇中被引入并维持。因此，pYT923S载体是一种有用的放线菌遗传工具，有望为今后放线菌生物学和致病性的研究提供帮助。

{"title":"Evaluation of Mycobacterium-derived plasmids for application in oral Actinomyces species","authors":"Sakiko Ohara , Yijuan Sheng , Yuki Nishiya , Ikue Tosa , Katsuki Takebe , Yuki Arimura , Hiroshi Mese , Naoko Ohara , Naoya Ohara","doi":"10.1016/j.job.2025.100718","DOIUrl":"10.1016/j.job.2025.100718","url":null,"abstract":"<div><h3>Objectives</h3><div>Genetic manipulation tools are essential for elucidating the pathogenic mechanisms of microorganisms. Several species of <em>Actinomyces,</em> including <em>A</em>. <em>israelii,</em> are present in the oral cavity and they are the causative agents of actinomycosis. However, efficient gene-editing tools for these species have not yet been developed. In this study, the aim was to evaluate the introduction of foreign genes into <em>Actinomyces</em> using plasmids derived from <em>Mycobacterium</em>, which belong to the same class as <em>Actinomycetes</em>.</div></div><div><h3>Methods</h3><div>A truncated derivative of pYT923, pYT923S, which contains the replication origin of the <em>M</em>. <em>scrofulaceum</em> plasmid pMSC262 was constructed and introduced into <em>A. israelii</em> by electrotransformation.</div></div><div><h3>Results</h3><div>pYT923S was successfully introduced into <em>A. israelii</em>. The transformation efficiency of <em>A. israelii</em> was approximately 7–66 CFU/μg of DNA, and all transformed colonies harbored pYT923S. The plasmid recovered from <em>A. israelii</em> replicated in <em>Escherichia coli</em>.</div></div><div><h3>Conclusions</h3><div>pYT923S was introduced into and maintained within <em>A. israelii</em>. Therefore, the pYT923S vector represents a useful genetic tool for <em>Actinomyces</em> and it is expected to facilitate future studies on the biology and pathogenicity of <em>Actinomyces</em>.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"68 1","pages":"Article 100718"},"PeriodicalIF":2.3,"publicationDate":"2025-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145691039","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Early senescence in salivary glands: A multimodal analysis across parotid, submandibular, and sublingual glands 唾液腺的早期衰老：一项跨腮腺、下颌骨和舌下腺的多模态分析

IF 2.3 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Journal of Oral Biosciences

Pub Date : 2025-11-29 DOI: 10.1016/j.job.2025.100717

Koichi Shibagaki , Kiyomi Ogawa , Tetsuro Horie , Ken-ichiro Kikuchi

Objectives

To elucidate the early-stage mechanisms of aging in salivary glands by focusing on the epigenetic regulator Jmjd3, proliferation marker proliferating cell nuclear antigen (PCNA), and senescence marker p16^INK4a in adult and middle-aged mice of both sexes.

Methods

Male and female C57BL/6JJcl mice at 18 weeks (adult) and 48 weeks (middle-aged) of age (n ≥ 5 per group) were used. The parotid, submandibular, and sublingual glands were collected, weighed, and analyzed by histology (hematoxylin and eosin, Alcian blue, periodic acid–Schiff staining), immunohistochemistry (for Jmjd3, PCNA, and p16^INK4a), quantitative image analysis, and real-time polymerase chain reaction. Statistical analysis was performed using analysis of variance or Kruskal–Wallis with appropriate post hoc tests (α = 0.05).

Results

Body weight increased with age, but the salivary gland weight relative to body weight remained unchanged. Histologically, at 48 weeks, the parotid and submandibular glands exhibited lymphocytic infiltration, whereas the sublingual glands did not. Jmjd3 was mainly localized in the acinar and striated duct cells and decreased with age in the parotid gland. PCNA-positive cells (acinar and ductal) were abundant at 18 weeks, but decreased after 48 weeks. p16^INK4a expression increased from 18 to 48 weeks, with initial staining in ductal cells and extension to some acinar cells by 48 weeks. p16^INK4a levels were higher in females than in males across all three glands.

Conclusions

By middle age, salivary glands exhibit early senescent changes, senescent cell accumulation, altered proliferative capacity, and shifts in Jmjd3-mediated epigenetic regulation, which occur in parallel with histological changes.

目的通过对成年和中年小鼠的表观遗传调控因子Jmjd3、增殖标志物增殖细胞核抗原（PCNA）和衰老标志物p16INK4a的研究，探讨唾液腺衰老的早期机制。方法采用18周龄（成年）和48周龄（中年）的C57BL/6JJcl雌雄小鼠（每组≥5只）。收集腮腺、下颌骨和舌下腺，称重，并进行组织学（苏木精和伊红、阿利新蓝、周期性酸希夫染色）、免疫组织化学（针对Jmjd3、PCNA和p16INK4a）、定量图像分析和实时聚合酶链反应分析。采用方差分析或Kruskal-Wallis进行统计学分析，并进行适当的事后检验（α = 0.05）。结果大鼠体重随年龄增长而增加，但唾液腺相对体重不变。组织学上，在48周时，腮腺和下颌腺表现出淋巴细胞浸润，而舌下腺则没有。Jmjd3主要分布于腮腺腺泡和纹管细胞，随着年龄的增长而减少。pcna阳性细胞（腺泡和导管）在18周时丰富，但在48周后减少。p16INK4a的表达从18周到48周增加，在导管细胞中开始染色，并在48周扩展到一些腺泡细胞。在所有三个腺体中，女性的p16INK4a水平都高于男性。结论人到中年，唾液腺出现早期衰老变化、衰老细胞积累、增殖能力改变以及jmjd3介导的表观遗传调控发生变化，这些变化与组织学改变同时发生。

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引用次数: 0