Journal of Oral Biosciences最新文献_第8页

Transcriptome and metabolome analyses of Streptococcus gordonii DL1 under acidic conditions 酸性条件下戈尔登链球菌 DL1 的转录组和代谢组分析。

IF 2.4 Q2 Dentistry

Journal of Oral Biosciences

Pub Date : 2024-03-01 DOI: 10.1016/j.job.2023.12.005

Naoto Hayashida , Yumiko Urano-Tashiro , Tetsuro Horie , Keitarou Saiki , Yuki Yamanaka , Yukihiro Takahashi

Objectives

Streptococcus gordonii is associated with the formation of biofilms, especially those that comprise dental plaque. Notably, S. gordonii DL1 causes infective endocarditis (IE). Colonization of this bacterium requires a mechanism that can tolerate a drop in environmental pH by producing acid via its own sugar metabolism. The ability to survive acidic environmental conditions might allow the bacterium to establish vegetative colonization even in the endocardium due to inflammation-induced lowering of pH, increasing the risk of IE. At present, the mechanism by which S. gordonii DL1 survives under acidic conditions is not thoroughly elucidated. The present study was thus conducted to elucidate the mechanism(s) by which S. gordonii DL1 survives under acidic conditions.

Methods

We analyzed dynamic changes in gene transcription and intracellular metabolites in S. gordonii DL1 exposed to acidic conditions, using transcriptome and metabolome analyses.

Results

Transcriptome analysis revealed upregulation of genes involved in heat shock response and glycolysis, and down regulation of genes involved in phosphotransferase systems and biosynthesis of amino acids. The most upregulated genes were a beta-strand repeat protein of unknown function (SGO_RS06325), followed by copper-translocating P-type ATPase (SGO_RS09470) and malic enzyme (SGO_RS01850). The latter two of these contribute to cytoplasmic alkalinization. S. gordonii mutant strains lacking each of these genes showed significantly reduced survival under acidic conditions. Metabolome analysis revealed that cytoplasmic levels of several amino acids were reduced.

Conclusions

S. gordonii survives the acidic conditions by recovering the acidic cytoplasm using the various activities, which are regulated at the transcriptional level.

目的：戈登链球菌与生物膜的形成有关，尤其是构成牙菌斑的生物膜。值得注意的是，戈登链球菌 DL1 会导致感染性心内膜炎（IE）。这种细菌的定殖需要一种机制，通过自身的糖代谢产生酸，从而耐受环境 pH 值的下降。该细菌在酸性环境条件下的生存能力可能会使其在炎症引起的 pH 值降低时也能在心内膜建立无性定植，从而增加 IE 的风险。目前，S. gordonii DL1 在酸性条件下的生存机制尚未得到彻底阐明。因此，本研究旨在阐明 S. gordonii DL1 在酸性条件下的生存机制：方法：我们使用转录组和代谢组分析方法分析了暴露于酸性条件下的 S. gordonii DL1 基因转录和细胞内代谢物的动态变化：结果：转录组分析显示，参与热休克反应和糖酵解的基因上调，参与磷酸转移酶系统和氨基酸生物合成的基因下调。上调最多的基因是功能不明的β链重复蛋白（SGO_RS06325），其次是铜转运P型ATP酶（SGO_RS09470）和苹果酸酶（SGO_RS01850）。后两者有助于细胞质碱化。缺乏这两个基因的 S. gordonii 突变菌株在酸性条件下的存活率明显降低。代谢组分析表明，几种氨基酸的细胞质水平降低：结论：戈尔登酵母菌在酸性条件下的存活是通过利用各种活动恢复酸性细胞质实现的，这些活动在转录水平上受到调控。

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引用次数: 0

Morphological differences between the first and second maxillary premolar crowns: A three-dimensional surface homologous modeling analysis 上颌第一和第二前磨牙牙冠的形态差异：三维表面同源建模分析。

IF 2.4 Q2 Dentistry

Journal of Oral Biosciences

Pub Date : 2024-03-01 DOI: 10.1016/j.job.2024.01.010

Julie Miyazaki , Shintaro Kondo , Toyohisa Tanijiri , Shinichi Negishi

Objectives

The current study used a three-dimensional (3D) surface homologous modeling to analyze the structure of maxillary first premolar (P¹) and second premolar (P²) crowns, to identify any morphological differences between them, particularly in their cuspal structures.

Methods

The study sample comprised 27 male elementary and junior high school students from Chiba Prefecture, Japan. Plaster casts were collected and the 3D coordinates were used to measure the crown structures. Thereafter, principal component (PC) analysis was carried out using the 3D coordinates of the homologous models, containing 4498 anatomical data points, including 9 landmarks.

Results

The findings indicated that P¹ was significantly larger than P², despite both teeth exhibiting similar intercuspal distances. The homologous model analysis revealed that 61.5 % of the total variance could be explained up to the fourth PC. Overall size and shape in the mesiodistal and buccolingual directions were estimated using PC1 and PC2, respectively. Both components highlighted a shape factor, indicating that the buccal cusp was more well-developed than the lingual cusp in P¹ compared to P².

Conclusions

The variations in the size of the mesial and distal premolar teeth and the relationships between the cusps in the completed tooth crowns can be explained using molecular biology developmental models.

研究目的本研究使用三维（3D）表面同源建模分析上颌第一前磨牙（P1）和第二前磨牙（P2）牙冠的结构，以确定它们之间的形态差异，尤其是尖牙结构的差异：研究样本包括来自日本千叶县的 27 名小学和初中男生。采集石膏模型并使用三维坐标测量牙冠结构。之后，使用同源模型的三维坐标进行主成分（PC）分析，其中包含 4498 个解剖数据点，包括 9 个地标：结果：研究结果表明，尽管两颗牙齿的齿间距离相似，但 P1 明显大于 P2。同源模型分析表明，61.5%的总方差可以用第四个PC来解释。中轴方向和颊舌方向的整体大小和形状分别由 PC1 和 PC2 估算。这两个成分都突出了一个形状因子，表明与 P2 相比，P1 中的颊尖比舌尖更发达：结论：分子生物学发育模型可以解释前臼齿中、远端的大小变化以及完整牙冠中尖牙之间的关系。

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引用次数: 0

Role of macrophages in trigeminal ganglia in ectopic orofacial pain associated with pulpitis 三叉神经节中的巨噬细胞在与牙髓炎相关的异位面痛中的作用。

IF 2.4 Q2 Dentistry

Journal of Oral Biosciences

Pub Date : 2024-03-01 DOI: 10.1016/j.job.2024.02.001

Miki Sunaga, Yoshiyuki Tsuboi, Akihiro Kaizu, Masamichi Shinoda

Objectives

This study aimed to elucidate the role of macrophages in the trigeminal ganglia (TG) in developing pulpitis-associated ectopic orofacial pain.

Methods

Rats underwent maxillary pulp exposure, and Fluoro-Gold (FG) was administered in the ipsilateral whisker pad (WP). Head withdrawal threshold (HWT) upon mechanical stimulation of the WP was recorded, and liposomal clodronate clophosome-A (LCCA; macrophage depletion agent) was administered to the TG at three and four days after pulp exposure. Immunohistochemically, TG sections were stained with anti-Iba1 (a macrophage marker) and anti-Nav1.7 antibodies.

Results

Pulp exposure decreased HWT and increased the number of Iba1-IR cells near FG-labelled TG neurons. LCCA inhibited the decrease in HWT and stopped the increase of FG-labelled Nav1.7-IR TG neurons in the pulpitis group.

Conclusions

Activation of macrophages by pulpitis induces the overexpression of Nav1.7 in TG neurons receiving inputs from WP, resulting in pulpitis-induced ectopic facial mechanical allodynia.

研究目的本研究旨在阐明三叉神经节（TG）中的巨噬细胞在发生牙髓炎相关异位性口腔疼痛中的作用：方法：对大鼠进行上颌牙髓暴露，并在同侧须垫（WP）注射氟金（FG）。记录WP受到机械刺激时的头部退缩阈值（HWT），并在牙髓暴露后的第三天和第四天在TG中施用氯膦酸脂质体clophosome-A（LCCA；巨噬细胞消耗剂）。用抗Iba1（一种巨噬细胞标记物）和抗Nav1.7抗体对TG切片进行免疫组化染色：结果：果肉暴露降低了HWT，增加了FG标记的TG神经元附近的Iba1-IR细胞数量。LCCA抑制了牙髓炎组HWT的下降，并阻止了FG标记的Nav1.7-IR TG神经元的增加：结论：牙髓炎激活巨噬细胞可诱导接受 WP 输入的 TG 神经元过量表达 Nav1.7，从而导致牙髓炎诱发的异位面部机械痛。

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引用次数: 0

Influence of IgA nephropathy on the progression of pulpitis and apical periodontitis in HIGA mice IgA肾病对HIGA小鼠牙髓炎和根尖牙周炎进展的影响。

IF 2.4 Q2 Dentistry

Journal of Oral Biosciences

Pub Date : 2024-03-01 DOI: 10.1016/j.job.2023.11.003

Reona Hayashi , Shiori Yamazaki , Noriko Mutoh , Tatsuo Hashimoto , Hayato Ohshima , Nobuyuki Tani-Ishii

Objectives

Immunoglobulin (Ig)A nephropathy has been associated with oral infections such as periodontitis, but its pathogenesis is not fully understood; no treatments exist. This study analyzes the influence of IgA nephropathy, an autoimmune disease, on the pathogenesis of pulpitis and apical periodontitis.

Methods

Two groups of mice were used in pulp infection experiments: high serum IgA nephropathy model mice (HIGA) and control mice (BALB/c). Histologic analyses of the pulp and apical periodontal tissues were performed on days 3, 5, 7, 14, and 28 following oral bacterial infection. The dynamics of odontoblasts, apoptotic cells, and IgA expression were analyzed using anti-Nestin, TUNEL, and anti-IgA staining, respectively.

Results

Inflammatory cells infiltrated the exposed pulp at day three in both groups and by 14 days, these cells had infiltrated from the pulp to the apical periodontal tissue. The area of necrotic pulp tissue increased significantly in the control group at seven days. Odontoblasts decreased from day three onwards and disappeared by 28 days in both groups. The number of apoptotic cells in the pulp and apical periodontal tissues was significantly higher in the experimental group at day 28. The experimental group exhibited a significant increase in IgA production in the pulp after 14 days. Bone resorption in the apical periodontal tissue was significantly decreased in the experimental group at day 28.

Conclusions

The results of this study suggest that IgA nephropathy may modulate the inflammatory response and sustain long-term biological defense responses in pulpitis and apical periodontitis in HIGA mice.

目的:免疫球蛋白(Ig)A肾病与牙周炎等口腔感染有关，但其发病机制尚不完全清楚;没有治疗方法。本研究分析了自身免疫性疾病IgA肾病对牙髓炎和根尖牙周炎发病机制的影响。方法:采用高血清IgA肾病模型小鼠(HIGA)和对照小鼠(BALB/c)两组进行牙髓感染实验。在口腔细菌感染后的第3、5、7、14和28天对牙髓和根尖牙周组织进行组织学分析。分别采用抗nestin染色、TUNEL染色和抗IgA染色分析成牙细胞、凋亡细胞和IgA表达的动态。结果:两组的炎症细胞在第3天浸润到暴露的牙髓，到第14天，这些细胞从牙髓浸润到根尖牙周组织。第7天，对照组牙髓坏死面积明显增加。两组成牙体细胞从第3天开始减少，并在第28天消失。第28天，实验组牙髓和根尖牙周组织中凋亡细胞数量明显增多。14天后，试验组牙髓中IgA产量显著增加。第28天，实验组根尖牙周组织骨吸收明显减少。结论:本研究结果提示IgA肾病可能调节HIGA小鼠牙髓炎和根尖牙周炎的炎症反应并维持长期的生物防御反应。

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引用次数: 0

Globo-series Gb4 activates ERK and promotes the proliferation of osteoblasts Globo系列Gb4激活ERK并促进成骨细胞增殖。

IF 2.4 Q2 Dentistry

Journal of Oral Biosciences

Pub Date : 2024-03-01 DOI: 10.1016/j.job.2023.10.004

Hanami Kato , Mayu Nagao , Koichi Furukawa , Yoshitaka Mishima , Shota Ichikawa , Takuma Sato , Ken Miyazawa , Kazunori Hamamura

Objectives

Globo-series Gb4 (globoside) is involved in the immune system and disease pathogenesis. We recently reported that systemic Gb4 deficiency in mice led to decreased bone formation due to a reduction in osteoblast number. However, it remains unclear whether Gb4 expressed in osteoblasts promotes their proliferation. Therefore, we investigated the role of Gb4 in osteoblast proliferation in vitro.

Methods

We examined osteoblast proliferation in Gb3 synthase knockout mice lacking Gb4. We investigated the effects of Gb4 synthase knockdown in the mouse osteoblast cell line MC3T3-E1 on its proliferation. Furthermore, we administered Gb4 to MC3T3-E1 cells in which Gb4 was suppressed by a glucosylceramide synthase (GCS) inhibitor and evaluated its effects on their proliferation. To elucidate the mechanisms by which Gb4 promotes osteoblast proliferation, the phosphorylated extracellular signal-regulated kinases 1 and 2 (ERK1/2) levels were measured in MC3T3-E1 cells.

Results

Osteoblast proliferation was lower in Gb3 synthase knockout mice lacking Gb4 than in wild-type mice. Proliferation was inhibited by Gb4 synthase knockdown in MC3T3-E1 cells. Furthermore, the administration of Gb4 to MC3T3-E1 cells, in which a GCS inhibitor suppressed Gb4, promoted their proliferation. Moreover, it increased the phosphorylated ERK1/2 levels in MC3T3-E1 cells.

Conclusions

Our results suggest that Gb4 expressed in osteoblasts promotes their proliferation through ERK1/2 activation.

目的：Globo系列Gb4（globoside）参与免疫系统和疾病的发病机制。我们最近报道，由于成骨细胞数量减少，小鼠全身性Gb4缺乏导致骨形成减少。然而，目前尚不清楚成骨细胞中表达的Gb4是否能促进其增殖。因此，我们研究了Gb4在体外成骨细胞增殖中的作用。方法：我们检测缺乏Gb4的Gb3合酶敲除小鼠的成骨细胞增殖。我们研究了小鼠成骨细胞系MC3T3-E1中Gb4合酶敲低对其增殖的影响。此外，我们将Gb4给予MC3T3-E1细胞，其中Gb4被葡萄糖神经酰胺合成酶（GCS）抑制剂抑制，并评估其对其增殖的影响。为了阐明Gb4促进成骨细胞增殖的机制，在MC3T3-E1细胞中测量了磷酸化的细胞外信号调节激酶1和2（ERK1/2）水平。结果：缺乏Gb4的Gb3合酶敲除小鼠的成骨细胞增殖低于野生型小鼠。Gb4合酶在MC3T3-E1细胞中被敲低抑制增殖。此外，向MC3T3-E1细胞施用Gb4（其中GCS抑制剂抑制Gb4）促进了它们的增殖。此外，它增加了MC3T3-E1细胞中磷酸化的ERK1/2水平。结论：我们的研究结果表明，成骨细胞中表达的Gb4通过ERK1/2的激活促进其增殖。

{"title":"Globo-series Gb4 activates ERK and promotes the proliferation of osteoblasts","authors":"Hanami Kato , Mayu Nagao , Koichi Furukawa , Yoshitaka Mishima , Shota Ichikawa , Takuma Sato , Ken Miyazawa , Kazunori Hamamura","doi":"10.1016/j.job.2023.10.004","DOIUrl":"10.1016/j.job.2023.10.004","url":null,"abstract":"<div><h3>Objectives</h3><p>Globo-series Gb4 (globoside) is involved in the immune system and disease pathogenesis. We recently reported that systemic Gb4 deficiency in mice led to decreased bone formation due to a reduction in osteoblast number. However, it remains unclear whether Gb4 expressed in osteoblasts promotes their proliferation. Therefore, we investigated the role of Gb4 in osteoblast proliferation <em>in vitro</em>.</p></div><div><h3>Methods</h3><p>We examined osteoblast proliferation in Gb3 synthase knockout mice lacking Gb4. We investigated the effects of Gb4 synthase knockdown in the mouse osteoblast cell line MC3T3-E1 on its proliferation. Furthermore, we administered Gb4 to MC3T3-E1 cells in which Gb4 was suppressed by a glucosylceramide synthase (GCS) inhibitor and evaluated its effects on their proliferation. To elucidate the mechanisms by which Gb4 promotes osteoblast proliferation, the phosphorylated extracellular signal-regulated kinases 1 and 2 (ERK1/2) levels were measured in MC3T3-E1 cells.</p></div><div><h3>Results</h3><p>Osteoblast proliferation was lower in Gb3 synthase knockout mice lacking Gb4 than in wild-type mice. Proliferation was inhibited by Gb4 synthase knockdown in MC3T3-E1 cells. Furthermore, the administration of Gb4 to MC3T3-E1 cells, in which a GCS inhibitor suppressed Gb4, promoted their proliferation. Moreover, it increased the phosphorylated ERK1/2 levels in MC3T3-E1 cells.</p></div><div><h3>Conclusions</h3><p>Our results suggest that Gb4 expressed in osteoblasts promotes their proliferation through ERK1/2 activation.</p></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1349007923001792/pdfft?md5=9a7f0f23e008ef2aec3886072bb8335e&pid=1-s2.0-S1349007923001792-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71522910","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Dynamic morphometric changes in the mandibular osteocytic lacunae of ovariectomized rats in response to teriparatide, as revealed by three-dimensional fluorescence analyses: Possible involvement of osteocytic perilacunar remodeling 三维荧光分析显示，去卵巢大鼠下颌骨骨细胞腔隙在特立帕肽作用下的动态形态变化:可能涉及骨细胞腔隙周围重构。

IF 2.4 Q2 Dentistry

Journal of Oral Biosciences

Pub Date : 2024-03-01 DOI: 10.1016/j.job.2023.11.010

Atsuko Nakanishi-Kimura , Aya Takakura , Marie Hoshi-Numahata , Haruhisa Watanabe , Mai Nishiura , Yoshiaki Sato , Ryoko Takao-Kawabata , Tadahiro Iimura

Objectives

Teriparatide [TPTD; human parathyroid hormone (hPTH1-34)] is an anti-osteoporotic drug with bone anabolic effects. Clinical and preclinical studies have indicated that TPTD has value in oral and maxillofacial bone therapies, including jawbone regeneration, periodontal tissue repair, and the treatment of medication-related osteonecrosis of the jaw. However, it is unclear whether the craniofacial bones respond to TPTD similarly to the axial and appendicular bones. Recent studies showed that TPTD acts on both osteocytes and osteoblasts. This study aimed to characterize distinct craniofacial bone sites, with a focus on morphometric changes in osteocytic lacunae in ovariectomized rats receiving TPTD.

Methods

Conventional bone histomorphometric analyses of mandibular and parietal bone sections were conducted. High-resolution confocal imaging-based three-dimensional fluorescence morphometric analyses of osteocytic lacunae in distinct mandibular and parietal bone sites were conducted.

Results

We observed dynamic changes in the morphometric characteristics of osteocytic lacunae specifically in alveolar and other mandibular bone sites upon TPTD administration.

Conclusions

These findings suggest that osteocytes in mandibular bone (specifically, alveolar bone) have unique functional characteristics of osteocytic perilacunar remodeling.

目的:特立帕肽;人甲状旁腺激素(hPTH)是一种具有骨合成代谢作用的抗骨质疏松药物。临床和临床前研究表明，TPTD在口腔颌面骨治疗中具有重要价值，包括颌骨再生、牙周组织修复和药物相关性颌骨骨坏死的治疗。然而，颅面骨对TPTD的反应是否与轴骨和阑尾骨相似尚不清楚。近年来的研究表明，TPTD同时作用于骨细胞和成骨细胞。本研究的目的是表征不同的颅面骨部位，重点是骨细胞腔隙的形态学变化在去卵巢大鼠接受TPTD。方法:对下颌骨和顶骨切片进行常规骨组织形态学分析。基于高分辨率共聚焦成像的三维荧光形态学分析骨细胞腔隙在不同的下颌骨和顶骨部位进行。结果:在给药后，我们观察到骨细胞腔隙的形态特征发生了动态变化，特别是在牙槽骨和其他颌骨部位。结论:这些发现提示下颌骨(特别是牙槽骨)的骨细胞具有独特的骨细胞腔旁重构功能特征。

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引用次数: 0

Ser252Trp mutation in fibroblast growth factor receptor 2 promotes branching morphogenesis in mouse salivary glands 成纤维细胞生长因子受体 2 的 Ser252Trp 突变促进了小鼠唾液腺的分支形态发生。

IF 2.4 Q2 Dentistry

Journal of Oral Biosciences

Pub Date : 2024-03-01 DOI: 10.1016/j.job.2024.01.001

Daiki Iwata , Kaori Kometani-Gunjigake , Kayoko Nakao-Kuroishi , Masahiro Mizuhara , Mitsushiro Nakatomi , Keiji Moriyama , Kentaro Ono , Tatsuo Kawamoto

Objectives

The purpose of this study was to perform morphological and immunohistochemical (IHC) analysis of the submandibular glands (SMGs) in early development in Apert syndrome model mice (Ap mice).

Methods

ACTB-Cre homozygous mice were mated with fibroblast growth factor receptor 2 (Fgfr2^+/Neo-S252W) mice; ACTB-Cre heterozygous mice (ACTB-Cre mice) at embryonic day (E) 13.5 served as the control group, and Fgfr2^+/S252W mice (Ap mice) served as the experimental group. Hematoxylin and eosin (H&E) staining was performed on SMGs; Total SMG area and epithelial area were determined, and the epithelial occupancy ratio was calculated. Immunostaining was performed to assess the localization of FGF signaling-related proteins. Next, bromodeoxyuridine (BrdU)-positive cells were evaluated to assess cell proliferation. Finally, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was performed to assess apoptosis in SMGs.

Results

The epithelial occupancy ratio was significantly higher in SMGs of Ap mice compared with that in SMGs of controls. FGF7 and bone morphogenetic protein 4 (BMP4) exhibited different localizations in SMGs of Ap mice compared with SMGs of controls. Cell proliferation was higher in SMGs of Ap mice compared with that of controls; however, apoptosis did not different significantly between the two groups.

Conclusion

Our results suggest that enhanced FGF signaling conferred by missense mutations in FGFR2 promotes branching morphogenesis in SMGs of Ap mice.

研究目的本研究的目的是对阿博特综合征模型小鼠（Ap小鼠）发育早期的下颌下腺（SMGs）进行形态学和免疫组化（IHC）分析：方法：将ACTB-Cre同源小鼠与成纤维细胞生长因子受体2（Fgfr2+/Neo-S252W）小鼠交配；以胚胎13.5天的ACTB-Cre杂合小鼠（ACTB-Cre小鼠）为对照组，Fgfr2+/S252W小鼠（Ap小鼠）为实验组。对SMG进行苏木精和伊红（H&E）染色；测定SMG总面积和上皮面积，并计算上皮占位比。进行免疫染色以评估 FGF 信号相关蛋白的定位。接着，对溴脱氧尿苷（BrdU）阳性细胞进行评价，以评估细胞增殖情况。最后，进行末端脱氧核苷酸转移酶 dUTP 缺口标记（TUNEL）染色，以评估 SMGs 的细胞凋亡情况：结果：与对照组相比，Ap小鼠SMG的上皮占位率明显升高。与对照组相比，FGF7和骨形态发生蛋白4（BMP4）在Ap小鼠SMG中的定位不同。与对照组相比，Ap小鼠SMGs中的细胞增殖率更高；然而，细胞凋亡在两组之间没有显著差异：我们的研究结果表明，FGFR2的错义突变增强了FGF信号传导，促进了Ap小鼠SMG的分支形态发生。

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引用次数: 0

Connective tissue growth factor enhances TGF-β1-induced osteogenic differentiation via activation of p38 MAPK in mesenchymal stem cells 结缔组织生长因子通过激活间充质干细胞中的 p38 MAPK 增强 TGF-β1 诱导的成骨分化。

IF 2.4 Q2 Dentistry

Journal of Oral Biosciences

Pub Date : 2024-03-01 DOI: 10.1016/j.job.2024.01.004

Hironori Yoshida , Seiji Yokota , Kazuro Satoh , Akira Ishisaki , Naoyuki Chosa

Objectives

Cellular differentiation is based on the effects of various growth factors. Transforming growth factor (TGF)-β1 plays a pivotal role in inducing osteogenic differentiation of mesenchymal stem cells (MSCs). In this study, we investigated the influence of connective tissue growth factor (CTGF), known to function synergistically with TGF-β1, on osteogenic differentiation in MSCs.

Methods

UE7T-13 cells were treated with TGF-β1 and/or CTGF. Subsequently, protein levels of intracellular signaling pathway molecules were determined through western blot analysis. The mRNA expression levels of osteogenic differentiation markers were investigated using reverse transcription-quantitative polymerase chain reaction. Bone matrix mineralization was evaluated through alizarin red staining.

Results

Co-treatment with TGF-β1 and CTGF resulted in the suppression of TGF-β1-induced phosphorylation of extracellular signal-regulated kinase 1/2, an intracellular signaling pathway molecule in MSCs, while significantly enhancing the phosphorylation of p38 mitogen-activated protein kinase (MAPK). In MSCs, co-treatment with CTGF and TGF-β1 led to increased expression levels of alkaline phosphatase and type I collagen, markers of osteogenic differentiation induced by TGF-β1. Osteopontin expression was observed only after TGF-β1 and CTGF co-treatment. Notably, bone sialoprotein and osteocalcin were significantly upregulated by treatment with CTGF alone. Furthermore, CTGF enhanced the TGF-β1-induced mineralization in MSCs, with complete suppression observed after treatment with a p38 MAPK inhibitor.

Conclusions

CTGF enhances TGF-β1-induced osteogenic differentiation and subsequent mineralization in MSCs by predominantly activating the p38 MAPK-dependent pathway.

目的：细胞分化基于各种生长因子的作用。转化生长因子（TGF）-β1 在诱导间充质干细胞（MSCs）的成骨分化中起着关键作用。本研究探讨了结缔组织生长因子（CTGF）对间充质干细胞成骨分化的影响：方法：用 TGF-β1 和/或 CTGF 处理 UE7T-13 细胞。方法：用 TGF-β1 和/或 CTGF 处理 UE7T-13 细胞，随后通过 Western 印迹分析测定细胞内信号通路分子的蛋白水平。使用反转录定量聚合酶链反应检测了成骨分化标志物的 mRNA 表达水平。通过茜素红染色评估骨基质矿化情况：结果：TGF-β1和CTGF联合处理可抑制TGF-β1诱导的间充质干细胞胞外信号调节激酶1/2（一种细胞内信号通路分子）的磷酸化，同时显著增强p38丝裂原活化蛋白激酶（MAPK）的磷酸化。在间充质干细胞中，CTGF 和 TGF-β1 的共同处理导致碱性磷酸酶和 I 型胶原蛋白的表达水平升高，而碱性磷酸酶和 I 型胶原蛋白是 TGF-β1 诱导成骨分化的标志物。只有在 TGF-β1 和 CTGF 联合处理后，才能观察到骨蛋白的表达。值得注意的是，单独使用 CTGF 处理后，骨硅蛋白和骨钙素的表达明显上调。此外，CTGF还能增强TGF-β1诱导的间充质干细胞矿化，在使用p38 MAPK抑制剂处理后可观察到完全抑制作用：结论：CTGF 主要通过激活 p38 MAPK 依赖性途径来增强 TGF-β1 诱导的间充质干细胞成骨分化和随后的矿化。

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引用次数: 0

Oral infection with Porphyromonas gingivalis augmented gingival epithelial barrier molecules alteration with aging 牙龈卟啉单胞菌口腔感染会随着年龄的增长而加剧牙龈上皮屏障分子的改变。

IF 2.4 Q2 Dentistry

Journal of Oral Biosciences

Pub Date : 2024-03-01 DOI: 10.1016/j.job.2024.01.012

Sarita Giri , Ayuko Takada , Durga Paudel , Osamu Uehara , Yoshihito Kurashige , Yasuhiro Kuramitsu , Masae Furukawa , Kenji Matsushita , Toshiya Arakawa , Toshiyuki Nagasawa , Yoshihiro Abiko , Yasushi Furuichi

Objective

Disruption of the gingival epithelial barrier is often mediated by aging or the pathogen Porphyromonas gingivalis. This study examined the combined effects of aging and P. gingivalis exposure on gingival epithelial barrier molecules.

Methods

In vitro experiments involved treating young- and senescence-induced primary human gingival epithelial progenitor cells (HGEPp) with P. gingivalis lipopolysaccharide (LPS). Transepithelial electrical resistance (TER) and paracellular permeability were measured. In vivo, male C57BL/6J mice aged 10 (young) and 80 (old) weeks were divided into four groups: young, old, young with P. gingivalis (Pg-Young) inoculation, and old with P. gingivalis (Pg-Old) inoculation. P. gingivalis was inoculated orally thrice a week for 5 weeks. The mice were sacrificed 30 days after the last inoculation, and samples were collected for further procedures. The junctional molecules (Claudin-1, Claudin-2, E-cadherin, and Connexin) were analyzed for mRNA expression using qRT-PCR and protein production using western blotting and immunohistochemistry. The alveolar bone loss and inflammatory cytokine levels in gingival tissues were also assessed.

Results

LPS-treated senescent cells exhibited a pronounced reduction in TER, increased permeability to albumin protein, significant upregulation of Claudin-1 and Claudin-2, and significant downregulation of E-cadherin and Connexin. Furthermore, the Pg-Old group showed identical results with aging in addition to an increase in alveolar bone loss, significantly higher than that in the other groups.

Conclusion

In conclusion, the host susceptibility to periodontal pathogens increases with age through changes in the gingival epithelial barrier molecules.

目的：牙龈上皮屏障的破坏通常是由衰老或病原体牙龈卟啉单胞菌引起的。本研究探讨了衰老和牙龈卟啉单胞菌暴露对牙龈上皮屏障分子的综合影响：体外实验包括用牙龈多糖（LPS）处理年轻和衰老诱导的原代人牙龈上皮祖细胞（HGEPp）。测量了跨上皮电阻（TER）的通透性。在体内，将年龄为 10 周（幼年）和 80 周（老年）的雄性 C57BL/6J 小鼠分为四组：幼年组、老年组、幼年接种牙龈脓疱病菌组（Pg-幼年组）和老年接种牙龈脓疱病菌组（Pg-老年组）。每周三次口服接种牙龈球菌，持续 5 周。小鼠在最后一次接种 30 天后被处死，并收集样本进行进一步处理。用 qRT-PCR 分析连接分子（Claudin-1、Claudin-2、E-cadherin 和 Connexin）的 mRNA 表达，用 Western 印迹和免疫组化分析蛋白质的生成。此外，还对牙龈组织中的牙槽骨损失和炎性细胞因子水平进行了评估：结果：经 LPS 处理的衰老细胞的 TER 明显降低，对白蛋白的通透性增加，Claudin-1 和 Claudin-2 明显上调，E-cadherin 和 Connexin 明显下调。此外，Pg-Old 组除了牙槽骨损失增加外，还表现出与老化相同的结果，明显高于其他组：总之，随着年龄的增长，宿主对牙周病原体的易感性会随着牙龈上皮屏障分子的变化而增加。

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引用次数: 0

Biological profiling using the human mandible 使用人类下颌骨进行生物分析。

IF 2.4 Q2 Dentistry

Journal of Oral Biosciences

Pub Date : 2024-03-01 DOI: 10.1016/j.job.2023.11.006

Doha Abualhija , Julieta Gómez García-Donas , Simon Shepherd , Scott McGregor , Ademir Franco , Scheila Manica

Background

In the absence of soft tissue, skeletal remains are analyzed to identify the deceased. This assessment involves establishing the biological profile that aids medicolegal investigations and fulfils the right of the dead to be identified. Since the mandible is the strongest bone in the skull and easily identifiable, even when fragmented, this study aimed to systematically review its value in constructing the biological profile in the published literature. We searched PubMed, ScienceDirect, Scopus, and Latin American and Caribbean Health Sciences Literature and collected cross-sectional studies published in English before 2021. A risk of bias assessment was completed based on Joanna Briggs Institute's critical appraisal tools. The data are presented descriptively and were analyzed using Microsoft Office Excel 365.

Highlight

Of the 104 eligible articles, 94 examined the sexual dimorphism of the mandible, while 25 attempted to estimate age. Ancestry and stature were the least explored biological characteristics (five and one articles, respectively). A metric analysis was the most common approach (n = 80), followed by morphological analysis and combined morphologic and metric techniques (n = 18 and n = 6, respectively). The results showed no statistically significant correlation between an individual's mandible and stature. Orthopantomogram radiography continues to be the most common radiographic technique for assessing the mandible.

Conclusion

The mandible is reliable when used for sex estimation; however, caution should be exercised in relying solely on it for morphological assessments. This review provides guidance on estimating age, sex, and ancestry directly from mandibular specimens or radiographs.

背景:在没有软组织的情况下，骨骼残骸被分析以识别死者。这一评估包括建立有助于法医调查和实现死者身份识别权利的生物学概况。由于下颌骨是颅骨中最坚固的骨骼，即使碎片化也很容易识别，因此本研究旨在系统地回顾其在已发表文献中构建生物学概况的价值。我们检索了PubMed、ScienceDirect、Scopus和拉丁美洲和加勒比健康科学文献，并收集了2021年之前发表的英文横断面研究。偏见风险评估基于乔安娜布里格斯研究所的关键评估工具完成。采用Microsoft Office Excel 365对数据进行了描述和分析。亮点:在104篇符合条件的文章中，94篇研究了下颌骨的两性二态，而25篇试图估计年龄。血统和身材是研究最少的生物学特征(分别为5篇和1篇)。计量分析是最常见的方法(n = 80)，其次是形态学分析和结合形态学和计量技术(n = 18和n = 6)。结果显示，一个人的下颌骨和身材之间没有统计学上的显著相关性。骨断层摄影仍然是评估下颌骨最常用的放射摄影技术。结论:下颌骨用于性别估计是可靠的;然而，在仅仅依靠它进行形态学评估时，应谨慎行事。这篇综述为直接从下颌标本或x线片估计年龄、性别和祖先提供了指导。

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引用次数: 0