Archives of oral biology最新文献_第8页

Verbascoside inhibits oral squamous cell carcinoma cell proliferation, migration, and invasion by the methyltransferase 3-mediated microRNA-31-5p/homeodomain interacting protein kinase 2 axis 马鞭草甙通过甲基转移酶 3 介导的 microRNA-31-5p/homeodomain 交互蛋白激酶 2 轴抑制口腔鳞状细胞癌细胞增殖、迁移和侵袭

IF 3 4区医学 Q1 Medicine

Archives of oral biology

Pub Date : 2024-04-22 DOI: 10.1016/j.archoralbio.2024.105979

Yuhua Huang, Wei Wu, Xing Zhang

Objective

The study aimed to investigate the effects of verbascoside on oral squamous cell carcinoma (OSCC) cellular behaviors and underlying molecular mechanisms.

Design

For this purpose, SCC9 and UM1 cell lines were treated with verbascoside, and their biological behaviors, including proliferation, migration, and invasion, were evaluated using cell counting kit-8, 5-Ethynyl-2′-deoxyuridine, and Transwell assays. The expression of methyltransferase-3 (METTL3), microRNA (miR)− 31–5p, and homeodomain interacting protein kinase-2 (HIPK2) were examined using quantitative real-time polymerase chain reaction (qRT-PCR). The interaction between METTL3 and miR-31–5p was evaluated by RNA immunoprecipitation and methylated RNA immunoprecipitation, while the interaction between miR-31–5p and HIPK2 was evaluated by dual-luciferase reporter analysis.

Results

The results indicated inhibition of OSCC cell proliferation, migration, and invasion post verbascoside treatment. Similarly, METTL3 was upregulated in OSCC cells and was inhibited post-verbascoside treatment. Overexpressing METTL3 promoted the cellular processes. Moreover, miR-31–5p was upregulated in OSCC cells, where METTL3 facilitated the processing of miR-31–5p in an N6-methyladenosine (m6A)-dependent manner. The HIPK2 served as miR-31–5p target, where overexpressing miR-31–5p or HIPK2 knockdown reversed the suppression of verbascoside-induced biological behaviors.

Conclusions

Verbascoside inhibited the progression of OSCC by inhibiting the METTL3-regulated miR-31–5p/HIPK2 axis. These findings suggested that verbascoside might be an effective drug for OSCC therapy.

目的本研究旨在探讨马鞭草苷对口腔鳞状细胞癌（OSCC）细胞行为的影响及其潜在的分子机制。为此，研究人员用马鞭草苷处理了SCC9和UM1细胞系，并用细胞计数试剂盒-8、5-乙炔基-2′-脱氧尿苷和Transwell试验评估了它们的生物学行为，包括增殖、迁移和侵袭。使用实时定量聚合酶链反应（qRT-PCR）检测了甲基转移酶-3（METTL3）、microRNA（miR）- 31-5p和同源结构域相互作用蛋白激酶-2（HIPK2）的表达。通过 RNA 免疫沉淀和甲基化 RNA 免疫沉淀评估了 METTL3 与 miR-31-5p 之间的相互作用，而通过双荧光素酶报告分析评估了 miR-31-5p 与 HIPK2 之间的相互作用。同样，METTL3 在 OSCC 细胞中上调，并在马鞭草苷处理后受到抑制。过表达 METTL3 会促进细胞进程。此外，miR-31-5p 在 OSCC 细胞中上调，其中 METTL3 以 N6-甲基腺苷（m6A）依赖的方式促进了 miR-31-5p 的处理。HIPK2是miR-31-5p的靶点，过表达miR-31-5p或敲除HIPK2可逆转对马鞭草苷诱导的生物学行为的抑制。这些研究结果表明，马鞭草苷可能是治疗OSCC的有效药物。

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引用次数: 0

TRIB3 promotes the growth of oral squamous cell carcinoma by regulating JNK/JUN-mediated aerobic glycolysis TRIB3 通过调节 JNK/JUN 介导的有氧糖酵解促进口腔鳞状细胞癌的生长

IF 3 4区医学 Q1 Medicine

Archives of oral biology

Pub Date : 2024-04-21 DOI: 10.1016/j.archoralbio.2024.105977

Zhaolun Meng , Yan Wang , Xiao Wang , Xuefeng Han

Objective

The potentiation of glycolysis is a leading driver of squamous cell carcinoma. Targeted modulation of the glycolytic process might be a pivotal tool for treating squamous cell carcinoma. Tribble homolog 3 (TRIB3) expression is elevated in some squamous cell carcinomas and correlates with poor prognosis. We investigated whether increased TRIB3 expression contributes to the progression of oral squamous cell carcinoma (OSCC) by modulating glycolysis.

Methods

We analyzed the expression of TRIB3 in the TCGA database for clinical tissue samples, in vitro, and in vivo. Cell proliferation, migration, invasion, and apoptosis were observed by overexpressing or knocking down TRIB3. Crucially, the impact of TRIB3 on aerobic glycolysis in OSCC was also probed in our study, including glucose uptake, lactate content, ATP production, extracellular acidification rate, and oxygen consumption rate. Importantly, we examined the relationship between TRIB3 and the JNK/JUN pathway and whether it regulates glycolytic processes in OSCC cells through the JNK/JUN pathway. Finally, tumor growth in vivo was tested using Xenograft models to observe the effect of knockdown TRIB3.

Results

Our study identified TRIB3 as the most variable and prognostic in OSCC. A significant high expression of TRIB3 in OSCC cells was determined in vitro and promoted cell proliferation, migration, invasion, apoptosis, and aerobic glycolysis. Knockdown of TRIB3 produced opposite effects. In addition, these effects are regulated by the JNK/JUN pathway. The use of JNK inhibitor inhibited the pro-growth and glycolytic effects of TRIB3 on OSCC cells. Finally, we further determined that TRIB3 knockdown would effectively suppress tumor growth in vivo.

Conclusion

This study reveals that TRIB3 promotes OSCC growth by regulating JNK/JUN pathway-mediated aerobic glycolysis, and TRIB3 may be a potential target for treating OSCC.

目的糖酵解的增强是鳞状细胞癌的主要驱动因素。靶向调节糖酵解过程可能是治疗鳞状细胞癌的关键手段。Tribble homolog 3（TRIB3）在一些鳞状细胞癌中表达升高，并与不良预后相关。我们研究了 TRIB3 表达的增加是否会通过调节糖酵解而导致口腔鳞状细胞癌（OSCC）的进展。通过过表达或敲除 TRIB3 观察了细胞的增殖、迁移、侵袭和凋亡。最重要的是，我们的研究还探究了TRIB3对OSCC有氧糖酵解的影响，包括葡萄糖摄取、乳酸含量、ATP产生、细胞外酸化率和氧消耗率。重要的是，我们研究了TRIB3与JNK/JUN通路之间的关系，以及它是否通过JNK/JUN通路调节OSCC细胞的糖酵解过程。最后，利用Xenograft模型检测了肿瘤在体内的生长情况，以观察敲除TRIB3的效果。体外研究发现，TRIB3 在 OSCC 细胞中的高表达可促进细胞增殖、迁移、侵袭、凋亡和有氧糖酵解。敲除 TRIB3 则会产生相反的效果。此外，这些效应受 JNK/JUN 通路的调控。使用 JNK 抑制剂可抑制 TRIB3 对 OSCC 细胞的促生长和糖酵解作用。结论本研究揭示了TRIB3通过调节JNK/JUN通路介导的有氧糖酵解促进OSCC生长，TRIB3可能是治疗OSCC的潜在靶点。

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引用次数: 0

Low-fluoride gels supplemented with nano-sized sodium trimetaphosphate reduce dentin erosive wear in vitro 添加了纳米级三偏磷酸钠的低氟凝胶可减少体外牙本质侵蚀磨损

IF 3 4区医学 Q1 Medicine

Archives of oral biology

Pub Date : 2024-04-15 DOI: 10.1016/j.archoralbio.2024.105973

Beatriz Díaz-Fabregat , Alberto Carlos Botazzo Delbem , Wilmer Ramírez-Carmona , Letícia Cabrera Capalbo , Liliana Carolina Báez-Quintero , Annette Wiegand , Douglas Roberto Monteiro , Juliano Pelim Pessan

Objective

The study assessed the effect of low-fluoride gels supplemented with micrometric or nano-sized sodium trimetaphosphate (TMP) on dentin erosive wear in vitro.

Design

Bovine dentin blocks (n = 154) were selected by surface microhardness and randomly allocated into seven groups (n = 22/group), according to the gels: Placebo; 4500 ppm F (4500F); 9000 ppm F (9000F); 5% TMP microparticulate plus 4500F (5TMPm+4500F); 2.5% TMP nanoparticulate plus 4500 F (2.5TMPn+4500F); 5% TMP nanoparticulate plus 4500F (5TMPn+4500F); and 12,300 ppm F acid gel (APF). All blocks were treated only once for 60 s and cyclically eroded (ERO, citric acid, 4 × 90 s/day) or eroded and brushed (4 × 15 s/day, five strokes/s, ERO+ABR) over five days (each subgroup n = 11). Dentin wear and integrated hardness loss in depth (ΔKHN) were determined, and the data were submitted to two-way ANOVA, followed by Tukey’s test, and Spearman’s correlation (p < 0.05).

Results

For ERO, all gels containing 4500F supplemented with TMP significantly reduced dentin wear compared with their counterpart without TMP, reaching values similar to 9000F. For ERO+ABR, 5TMPn+ 4500F gel led to significantly lower wear than all its counterparts, reaching values similar to 9000F and APF. As for ΔKHN, all gels containing TMP promoted superior protective effects compared with 4500F, reaching values similar to 9000F and APF under both challenges. A positive correlation between dentin wear and mineral content in depth was verified.

Conclusions

Gels containing 4500F supplemented with TMP significantly reduced dentin erosive wear compared with pure 4500F, with additional benefit from the use of nanoparticles.

设计根据表面显微硬度选择牛牙本质块（n = 154），并根据凝胶随机分配到七个组（n = 22/组）：安慰剂组；4500 ppm F（4500F）组；9000 ppm F（9000F）组；5% TMP 微颗粒加 4500 F（5TMPm+4500F）组；2.5% TMP 纳米颗粒加 4500 F（2.5TMPn+4500F）组；5% TMP 纳米颗粒加 4500 F（5TMPn+4500F）组；以及 12,300 ppm F 酸凝胶（APF）组。所有牙块只处理一次，每次 60 秒，然后在五天内循环腐蚀（ERO，柠檬酸，4 × 90 秒/天）或腐蚀和刷洗（4 × 15 秒/天，5 次/秒，ERO+ABR）（每个亚组 n = 11）。测定牙本质磨损和深度综合硬度损失（ΔKHN），并对数据进行双向方差分析，然后进行 Tukey 检验和 Spearman 相关性检验（p < 0.05）。对于ERO+ABR，5TMPn+4500F凝胶的磨损程度明显低于所有同类凝胶，磨损值与9000F和APF相似。至于 ΔKHN，与 4500F 相比，所有含有 TMP 的凝胶都能产生更好的保护效果，在两种挑战下都能达到与 9000F 和 APF 相似的值。结论与纯 4500F 相比，含有 TMP 的 4500F 凝胶能显著减少牙本质的侵蚀性磨损，纳米颗粒的使用还能带来额外的益处。

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引用次数: 0

4-hydroxy-3-methoxybenzaldehyde causes attrition of biofilm formation and quorum sensing–associated virulence factors of Streptococcus mutans 4-hydroxy-3-methoxybenzaldehyde 可破坏变异链球菌的生物膜形成和与法定量感应相关的毒力因子

IF 3 4区医学 Q1 Medicine

Archives of oral biology

Pub Date : 2024-04-15 DOI: 10.1016/j.archoralbio.2024.105976

Pitchaipillai Sankar Ganesh

Objective

The present study investigated the effects of 4-hydroxy-3-methoxybenzaldehyde (4-H-3-MB) against Streptococcus mutans (S. mutans) using an in vitro cariogenic biofilm model.

Design

The antimicrobial susceptibility of biofilm-forming S. mutans was evaluated by disc diffusion method. In vitro investigations were performed using crystal violet staining assay (biofilm assay), exopolysaccharide (EPS) assay, acid production, growth curve analysis, optical microscopic, and FE-SEM analyses to determine the antibiofilm activity of 4-H-3-MB.

Results

S. mutans (SDC-05) was resistant to ampicillin, piperacillin/tazobactam and ceftriaxone, whereas the other strains of S. mutans (SDC-01, 02, 03 and SDC-04) were sensitive to all the antibiotics tested. 4-H-3-MB showed promising antibiofilm activity on S. mutans UA159 (79.81 %, 67.76 % and 56.31 %) and S. mutans SDC-05 (77.00 %, 59.48 % and 48.22 %) at the lowest concentration of 0.2, 0.1, 0.05 mg/ml. 4-H-3-MB did not inhibit bacterial growth even at concentrations 0.2 mg/ml. Similarly, 4-H-3-MB led to significant attrition in exopolysaccharide (EPS) and acid production by S. mutans UA159 and S. mutans (SDC-05) at the concentration of 0.2, 0.1 mg/ml, respectively. Optical microscopy and FE-SEM analysis 4-H-3-MB reduced the biofilm thickness of S. mutans UA159 and S. mutans SDC-05 relative to the untreated specimens.

Conclusion

4-H-3-MB significantly inhibited biofilm formation by S. mutans in a dose-dependent manner. Hence, our findings indicate that the active principle of 4-H-3-MB could be used as a biofilm inhibiting agent against S. mutans.

本研究使用体外致龋生物膜模型研究了 4-羟基-3-甲氧基苯甲醛（4-H-3-MB）对变异链球菌（S. mutans）的作用。采用水晶紫染色检测法（生物膜检测法）、外多糖（EPS）检测法、产酸法、生长曲线分析法、光学显微镜和 FE-SEM 分析法进行体外研究，以确定 4-H-3-MB 的抗生物膜活性。结果 S. mutans（SDC-05）对氨苄西林、哌拉西林/他唑巴坦和头孢曲松耐药，而其他菌株（SDC-01、02、03 和 SDC-04）对所有测试的抗生素都敏感。在 0.2、0.1 和 0.05 毫克/毫升的最低浓度下，4-H-3-MB 对变异杆菌 UA159（79.81%、67.76% 和 56.31%）和变异杆菌 SDC-05 （77.00%、59.48% 和 48.22%）具有良好的抗生物膜活性。即使浓度为 0.2 毫克/毫升，4-H-3-MB 也不能抑制细菌的生长。同样，在 0.2、0.1 毫克/毫升的浓度下，4-H-3-MB 会导致变异杆菌 UA159 和变异杆菌（SDC-05）的外多糖（EPS）和产酸量显著减少。光学显微镜和 FE-SEM 分析显示，相对于未处理的标本，4-H-3-MB 可降低变异杆菌 UA159 和变异杆菌 SDC-05 的生物膜厚度。因此，我们的研究结果表明，4-H-3-MB 的活性成分可用作抑制变异沙雷氏菌形成生物膜的药物。

{"title":"4-hydroxy-3-methoxybenzaldehyde causes attrition of biofilm formation and quorum sensing–associated virulence factors of Streptococcus mutans","authors":"Pitchaipillai Sankar Ganesh","doi":"10.1016/j.archoralbio.2024.105976","DOIUrl":"https://doi.org/10.1016/j.archoralbio.2024.105976","url":null,"abstract":"<div><h3>Objective</h3><p>The present study investigated the effects of 4-hydroxy-3-methoxybenzaldehyde (4-H-3-MB) against <em>Streptococcus mutans</em> (<em>S. mutans</em>) using an in vitro cariogenic biofilm model.</p></div><div><h3>Design</h3><p>The antimicrobial susceptibility of biofilm-forming <em>S. mutans</em> was evaluated by disc diffusion method. <em>In vitro</em> investigations were performed using crystal violet staining assay (biofilm assay), exopolysaccharide (EPS) assay, acid production, growth curve analysis, optical microscopic, and FE-SEM analyses to determine the antibiofilm activity of 4-H-3-MB.</p></div><div><h3>Results</h3><p><em>S. mutans</em> (SDC-05) was resistant to ampicillin, piperacillin/tazobactam and ceftriaxone, whereas the other strains of <em>S. mutans</em> (SDC-01, 02, 03 and SDC-04) were sensitive to all the antibiotics tested. 4-H-3-MB showed promising antibiofilm activity on <em>S. mutans</em> UA159 (79.81 %, 67.76 % and 56.31 %) and <em>S. mutans</em> SDC-05 (77.00 %, 59.48 % and 48.22 %) at the lowest concentration of 0.2, 0.1, 0.05 mg/ml. 4-H-3-MB did not inhibit bacterial growth even at concentrations 0.2 mg/ml. Similarly, 4-H-3-MB led to significant attrition in exopolysaccharide (EPS) and acid production by <em>S. mutans</em> UA159 and <em>S. mutans</em> (SDC-05) at the concentration of 0.2, 0.1 mg/ml, respectively. Optical microscopy and FE-SEM analysis 4-H-3-MB reduced the biofilm thickness of <em>S. mutans</em> UA159 and <em>S. mutans</em> SDC-05 relative to the untreated specimens.</p></div><div><h3>Conclusion</h3><p>4-H-3-MB significantly inhibited biofilm formation by <em>S. mutans</em> in a dose-dependent manner<em>.</em> Hence, our findings indicate that the active principle of 4-H-3-MB could be used as a biofilm inhibiting agent against <em>S. mutans</em>.</p></div>","PeriodicalId":8288,"journal":{"name":"Archives of oral biology","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140606705","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A prognostic model built on amino acid metabolism patterns in HPV-associated head and neck squamous cell carcinoma 基于人乳头瘤病毒相关头颈部鳞状细胞癌氨基酸代谢模式的预后模型

IF 3 4区医学 Q1 Medicine

Archives of oral biology

Pub Date : 2024-04-15 DOI: 10.1016/j.archoralbio.2024.105975

Fengyang Jing , Lijing Zhu , Jiaying Bai , Xuan Zhou , Lisha Sun , Heyu Zhang , Tiejun Li

Objectives

To compare amino acid metabolism patterns between HPV-positive and HPV-negative head and neck squamous cell carcinoma (HNSCC) patients and identify key genes for a prognostic model.

Design

Utilizing the Cancer Genome Atlas dataset, we analyzed amino acid metabolism genes, differentiated genes between HPV statuses, and selected key genes via LASSO regression for the prognostic model. The model's gene expression was verified through immunohistochemistry in clinical samples. Functional enrichment and CIBERSORTx analyses explored biological functions, molecular mechanisms, and immune cell correlations. The model's prognostic capability was assessed using nomograms, calibration, and decision curve analysis.

Results

We identified 1157 key genes associated with amino acid metabolism in HNSCC and HPV status. The prognostic model, featuring genes like IQCN, SLC22A1, SYT12, and TLX3, highlighted functions in development, metabolism, and pathways related to receptors and enzymes. It significantly correlated with immune cell infiltration and outperformed traditional staging in prognosis prediction, despite immunohistochemistry results showing limited clinical identification of HPV-related HNSCC.

Conclusions

Distinct amino acid metabolism patterns differentiate HPV-positive from negative HNSCC patients, underscoring the prognostic model's utility in predicting outcomes and guiding therapeutic strategies.

目的比较HPV阳性和HPV阴性头颈部鳞状细胞癌（HNSCC）患者的氨基酸代谢模式，并确定预后模型的关键基因。设计利用癌症基因组图谱数据集，我们分析了氨基酸代谢基因，区分了不同HPV状态的基因，并通过LASSO回归筛选出预后模型的关键基因。该模型的基因表达通过临床样本的免疫组化进行了验证。功能富集和 CIBERSORTx 分析探讨了生物功能、分子机制和免疫细胞相关性。结果我们发现了 1157 个与 HNSCC 中氨基酸代谢和 HPV 状态相关的关键基因。该预后模型以 IQCN、SLC22A1、SYT12 和 TLX3 等基因为特色，突出了其在发育、代谢以及与受体和酶相关的通路中的功能。尽管免疫组化结果显示 HPV 相关 HNSCC 的临床识别率有限，但该模型与免疫细胞浸润有明显相关性，在预后预测方面优于传统分期方法。

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引用次数: 0

GNAI3 mediated by Lin28A regulates lipopolysaccharide-induced inflammation and osteogenic differentiation in periodontal stem cells by mediating the NF-κB/NLRP3 inflammasome pathway 由Lin28A介导的GNAI3通过调解NF-κB/NLRP3炎性体通路调节脂多糖诱导的牙周干细胞炎症和成骨分化

IF 3 4区医学 Q1 Medicine

Archives of oral biology

Pub Date : 2024-04-12 DOI: 10.1016/j.archoralbio.2024.105974

Ling Guo, Hua Sun, Jiao Pu

Objectives

The aim of this study was to investigate the regulatory role of G protein subunit alpha i3 (GNAI3) in periodontitis.

Design

Following the induction of human periodontal ligament stem cells (hPDLSCs) with lipopolysaccharide (LPS), the mRNA and protein expressions of GNAI3 and Lin28A were detected by real-time quantitative polymerase chain reaction (RT-qPCR) and western blot. The transfection efficiency of Oe-GNAI3 and sh-Lin28A was examined by virtue of RT-qPCR and western blot. With the application of ELISA and flow cytometry, the releases of inflammatory cytokines and cell apoptosis were appraised. Alkaline phosphatase (ALP) staining and alizarin red S (ARS) staining were conducted to evaluate osteogenic differentiation. Next, the binding ability of Lin28A with GNAI3 mRNA was estimated by radioimmunoprecipitation (RIP) assay while the stability of GNAI3 mRNA was assessed utilizing RT-qPCR. Western blot was employed for the measurement of inflammation-, apoptosis- and nuclear factor-kappaB (NF-κB)/NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome pathway-related proteins and osteogenic markers.

Results

The expression of GNAI3 was down-regulated in LPS-induced hPDLSCs. After the transfection with Oe-GNAI3, the inflammation and apoptosis in LPS-induced hPDLSCs were inhibited while osteogenic differentiation was promoted. Moreover, Lin28A could stabilize GNAI3 mRNA and Lin28A knockdown significantly reduced GNAI3 expression. Further experiments verified that the inhibitory effects of GNAI3 overexpression on LPS-induced cellular inflammation and cell apoptosis as well as the promotive effects on osteogenic differentiation in hPDLSCs were all partially counteracted by Lin28A depletion, which may possibly be mediated via the regulation of the NF-κB/NLRP3 inflammasome pathway.

Conclusion

GNAI3 that mediated by Lin28A regulates the inflammation and osteogenic differentiation in LPS-induced hPDLSCs by mediating the NF-κB/NLRP3 inflammasome pathway.

设计用脂多糖（LPS）诱导人牙周韧带干细胞（hPDLSCs）后，通过实时定量聚合酶链式反应（RT-qPCR）和Western印迹检测GNAI3和Lin28A的mRNA和蛋白表达。通过 RT-qPCR 和 western 印迹检测了 Oe-GNAI3 和 sh-Lin28A 的转染效率。应用酶联免疫吸附和流式细胞术评估了炎症细胞因子的释放和细胞凋亡。碱性磷酸酶（ALP）染色和茜素红 S（ARS）染色评估了成骨分化。然后，通过放射免疫沉淀（RIP）试验评估了 Lin28A 与 GNAI3 mRNA 的结合能力，并利用 RT-qPCR 评估了 GNAI3 mRNA 的稳定性。采用 Western 印迹法测定炎症、细胞凋亡、核因子-卡巴（NF-κB）/类NOD受体家族含吡咯啉结构域3（NLRP3）炎性组通路相关蛋白和成骨标志物。转染 Oe-GNAI3 后，LPS 诱导的 hPDLSCs 的炎症和细胞凋亡得到抑制，成骨分化得到促进。此外，Lin28A能稳定GNAI3 mRNA，Lin28A敲除能显著降低GNAI3的表达。进一步的实验验证了过表达 GNAI3 对 LPS 诱导的细胞炎症和细胞凋亡的抑制作用，以及对 hPDLSCs 成骨分化的促进作用都被 Lin28A 损伤部分抵消，这可能是通过调控 NF-κB/NLRP3 炎性体通路介导的。结论 Lin28A介导的GNAI3通过介导NF-κB/NLRP3炎性体通路调节LPS诱导的hPDLSCs的炎症和成骨分化。

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引用次数: 0

The role of the Piezo1 channel in osteoblasts under cyclic stretching: A study on osteogenic and osteoclast factors 循环拉伸条件下 Piezo1 通道在成骨细胞中的作用：对成骨和破骨因子的研究

IF 3 4区医学 Q1 Medicine

Archives of oral biology

Pub Date : 2024-04-09 DOI: 10.1016/j.archoralbio.2024.105963

Ting Kang , Ziyuan Yang , Mengqi Zhou , Yanhua Lan , Yaya Hong , Xinyi Gong , Yongjia Wu , Min Li , Xuepeng Chen , Weifang Zhang

Objectives

Orthodontic tooth movement is a mechanobiological reaction induced by appropriate forces, including bone remodeling. The mechanosensitive Piezo channels have been shown to contribute to bone remodeling. However, information about the pathways through which Piezo channels affects osteoblasts remains limited. Thus, we aimed to investigate the influence of Piezo1 on the osteogenic and osteoclast factors in osteoblasts under mechanical load.

Materials and methods

Cyclic stretch (CS) experiments on MC3T3-E1 were conducted using a BioDynamic mechanical stretching device. The Piezo1 channel blocker GsMTx4 and the Piezo1 channel agonist Yoda1 were used 12 h before the application of CS. MC3T3-E1 cells were then subjected to 15% CS, and the expression of Piezo1, Piezo2, BMP-2, OCN, Runx2, RANKL, p-p65/p65, and ALP was measured using quantitative real-time polymerase chain reaction, western blot, alkaline phosphatase staining, and immunofluorescence staining.

Results

CS of 15% induced the highest expression of Piezo channel and osteoblast factors. Yoda1 significantly increased the CS-upregulated expression of Piezo1 and ALP activity but not Piezo2 and RANKL. GsMTx4 downregulated the CS-upregulated expression of Piezo1, Piezo2, Runx2, OCN, p-65/65, and ALP activity but could not completely reduce CS-upregulated BMP-2.

Conclusions

The appropriate force is more suitable for promoting osteogenic differentiation in MC3T3-E1. The Piezo1 channel participates in osteogenic differentiation of osteoblasts through its influence on the expression of osteogenic factors like BMP-2, Runx2, and OCN and is involved in regulating osteoclasts by influencing phosphorylated p65. These results provide a foundation for further exploration of osteoblast function in orthodontic tooth movement.

目的正畸牙齿移动是一种由适当力量诱发的机械生物学反应，包括骨重塑。对机械敏感的压电通道已被证明有助于骨重塑。然而，有关压电通道影响成骨细胞的途径的信息仍然有限。因此，我们旨在研究机械负荷下 Piezo1 对成骨细胞中的成骨和破骨细胞因子的影响。在施加 CS 前 12 小时使用 Piezo1 通道阻断剂 GsMTx4 和 Piezo1 通道激动剂 Yoda1。然后将 MC3T3-E1 细胞置于 15% 的 CS 中，使用实时定量聚合酶链反应、Western 印迹、碱性磷酸酶染色和免疫荧光染色等方法检测 Piezo1、Piezo2、BMP-2、OCN、Runx2、RANKL、p-p65/p65 和 ALP 的表达。Yoda1 能明显提高 CS 上调的 Piezo1 和 ALP 活性的表达，但不能提高 Piezo2 和 RANKL 的表达。GsMTx4 下调了 CS 上调的 Piezo1、Piezo2、Runx2、OCN、p-65/65 和 ALP 活性的表达，但不能完全降低 CS 上调的 BMP-2。Piezo1 通道通过影响 BMP-2、Runx2 和 OCN 等成骨因子的表达参与成骨细胞的成骨分化，并通过影响磷酸化 p65 参与破骨细胞的调控。这些结果为进一步探索成骨细胞在正畸牙齿移动中的功能奠定了基础。

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引用次数: 0

Antimicrobial action and cytotoxicity of hypochlorous acid obtained from an innovative electrolytic device – An in vitro study 从创新电解装置中获得的次氯酸的抗菌作用和细胞毒性--体外研究

IF 3 4区医学 Q1 Medicine

Archives of oral biology

Pub Date : 2024-04-08 DOI: 10.1016/j.archoralbio.2024.105966

Matheus Albino Souza , Mylena Lazareti Zanella , Gabriele Nichetti Vanin , Felipe Gomes Dallepiane , Camila Yasmin Monteiro Pizzi , Eduarda Rizzon Ferreira , Marciele Cristiane Spanenberg Fuhr , Nathan Mateus Piccolo , Huriel Scartazzini Palhano , Jordana da Silva Koch , Kellyn Rocca Souza , Ubirajara Maciel da Costa , Vanessa Valgas dos Santos , Liviu Steier , Charise Dallazem Bertol , José Antônio Poli de Figueiredo

Objective

This study evaluated the antimicrobial effect and cytotoxicity of hypochlorous acid(HClO) obtained from an innovative electrolytic device.

Design

The root canals of fifty extracted human teeth were inoculated with Enterococcus faecalis and divided into 5 groups (n = 10): DW (control); 2% chlorhexidine gel(CHX); 2.5% sodium hypochlorite(NaOCl); 250 ppm HClO and 500 ppm HClO. The counting of colony forming units evaluated the decontamination potential of each group. Cytotoxicity was evaluated after inoculation of tested protocols in fibroblastic cells for 3 min, calculating the cell viability. Specific statistical analysis was performed (α = 5%).

Results

The highest bacterial reduction was observed in experimental groups, with no statistical differences from each other (p > 0.05). The highest number of viable cells was observed in control group, followed by 250 ppm HClO and 500 ppm HClO groups, with statistical differences from each other (p < 0.05).

Conclusions

It could be concluded that HClO presented high antimicrobial activity and low cytotoxicity at both tested concentrations.

设计将 50 颗人类拔牙的根管接种粪肠球菌，并分为 5 组（n = 10）：DW（对照组）；2% 洗必泰凝胶（CHX）；2.5% 次氯酸钠（NaOCl）；250 ppm HClO 和 500 ppm HClO。菌落形成单位计数评估了各组的去污潜力。在成纤维细胞中接种测试方案 3 分钟后，对细胞毒性进行评估，计算细胞存活率。进行了具体的统计分析（α = 5%）。结果实验组的细菌减少量最高，但各组之间无统计学差异（p > 0.05）。对照组的存活细胞数最多，其次是 250 ppm HClO 组和 500 ppm HClO 组，各组之间存在统计学差异（p < 0.05）。

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引用次数: 0

Mechanism and functional verification of genes by virulence factors of P. gingivalis in ferroptosis 牙龈脓毒性因子在铁败血症中的作用机制和功能验证

IF 3 4区医学 Q1 Medicine

Archives of oral biology

Pub Date : 2024-04-07 DOI: 10.1016/j.archoralbio.2024.105965

Xinzhu Li, Ting Chen, Yinyu Fu, Bo Yang, Xiaoyu Lin, Jin Hou, Xiaojun Yang

Objective

Porphyromonas gingivalis (P. gingivalis) is a key etiological agent in periodontitis and functions as a facultative intracellular microorganism and involves many virulence factors. These virulence factors participate in multiple intracellular processes, like ferroptosis, the mechanistic underpinnings remain to be elucidated. Aim of this study was to investigate the effects of virulence factors on the host cells.

Design

Human umbilical vein endothelial cells (HUVECs) were treated with 4% paraformaldehyde-fixed P. gingivalis, and subsequent alterations in gene expression were profiled via RNA-seq. Further, the molecules associated with ferroptosis were quantitatively analyzed using qRT-PCR and Western blot.

Results

A total of 1125 differentially expressed genes (DEGs) were identified, encompassing 225 upregulated and 900 downregulated. Ferroptosis was conspicuously represented in the kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis, with notable upregulation of Heme oxygenase 1 (HMOX1), Ferritin light chain (FTL), and Solute carrier family 3 member 2 (SLC3A2) and downregulation of Scavenger receptor class A member 5 (SCARA5) and glutaminase (GLS). Random selection of DEGs for validation through qRT-PCR corroborated the RNA-Seq data (R² = 0.93). Kelch like ECH associated protein 1 (Keap1) protein expression decreased after 4 and 8 h, while NFE2 like bZIP transcription factor 2 (Nrf2) and HMOX1 were elevated, with significant nuclear translocation of Nrf2.

Conclusions

The virulence factors of P. gingivalis may potentially instigating ferroptosis through activation of the Keap1-Nrf2-HMOX1 signaling cascade, in conjunction with modulating the expression of other ferroptosis-associated elements. Further research is necessary to achieve a thorough comprehension of these complex molecular interactions.

目的牙龈卟啉单胞菌（P. gingivalis）是牙周炎的主要病原体，它是一种兼性细胞内微生物，涉及许多毒力因子。这些毒力因子参与了多种细胞内过程，如铁跃迁，但其机理基础仍有待阐明。本研究旨在研究毒力因子对宿主细胞的影响。设计用 4% 多聚甲醛固定的牙龈脓毒性噬菌体处理人脐静脉内皮细胞（HUVECs），并通过 RNA-seq 分析随后的基因表达变化。结果共鉴定出 1125 个差异表达基因（DEGs），包括 225 个上调基因和 900 个下调基因。在京都基因和基因组百科全书（KEGG）的富集分析中，铁蛋白沉积症的表现非常明显，血红素加氧酶 1（HMOX1）、铁蛋白轻链（FTL）和溶质运载家族 3 成员 2（SLC3A2）明显上调，而清道夫受体 A 类成员 5（SCARA5）和谷氨酰胺酶（GLS）则明显下调。通过 qRT-PCR 随机选择 DEGs 进行验证证实了 RNA-Seq 数据（R2 = 0.93）。结论牙龈脓毒性因子可能通过激活 Keap1-Nrf2-HMOX1 信号级联以及调节其他铁变态反应相关因子的表达来诱导铁变态反应。要彻底理解这些复杂的分子相互作用，还需要进一步的研究。

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引用次数: 0

Small extracellular vesicles derived from lipopolysaccharide-preconditioned dental follicle cells inhibit cell apoptosis and alveolar bone loss in periodontitis 从脂多糖预处理的牙泡细胞中提取的小细胞外囊泡可抑制牙周炎患者的细胞凋亡和牙槽骨丧失

IF 3 4区医学 Q1 Medicine

Archives of oral biology

Pub Date : 2024-04-02 DOI: 10.1016/j.archoralbio.2024.105964

Yanli Huang , Mujia Li , Qian Liu , Lu Song , Qianting Wang , Peihui Ding , Weidong Tian , Shujuan Guo

Objective

This study aimed to explore the effects of small extracellular vesicles derived from lipopolysaccharide-preconditioned dental follicle cells (L-D-sEV) on periodontal ligament cells from periodontitis affected teeth (p-PDLCs) in vitro and experimental periodontitis in mice.

Design

In vitro, the biological function of p-PDLCs and the underlying molecular mechanism were investigated by flow cytometry, Western blot, and quantitative real-time PCR (qRT-PCR) analysis. Eighteen-eight-week-old male C57BL/6 mice were randomly divided into three groups: control (Con), periodontitis (Peri), and L-D-sEV groups. Mice periodontitis model was induced by placing the 5–0 silk thread (around the maxillary second molar) and P.gingivalis (1 ×10⁷ CFUs per mouse). In vivo, the alveolar bone loss, osteoclast activity, and macrophage polarization were measured by micro-computed tomography and histological analysis.

Results

In vitro, the RANKL/OPG ratio and phosphorylation of JNK and P38 protein levels of p-PDLCs were significantly decreased after L-D-sEV administration. Besides, flow cytometry and qRT-PCR analysis showed that L-D-sEV reduced apoptosis of p-PDLCs, down-regulated apoptosis-related genes Caspase-3 and BCL-2-Associated X expression, and up-regulated B-cell lymphoma-2 gene levels. In vivo, L-D-sEV administration significantly reduced alveolar bone loss, inhibited osteoclast activity, and induced M2 polarization. The histological analysis showed that iNOS/CD206, RANKL/OPG, p-JNK/JNK, and p-P38/P38 ratios were significantly lower in the L-D-sEV group than in the Peri group.

Conclusions

L-D-sEV administration alleviated alveolar bone loss by mediating RANKL/OPG-related osteoclast activity and M2 macrophage polarization, alleviating p-PDLCs apoptosis and proliferation via the JNK and P38 pathways.

本研究旨在探讨脂多糖预处理的牙周滤泡细胞（L-D-sEV）衍生的小细胞外囊泡对体外牙周炎患牙牙周韧带细胞（p-PDLCs）和小鼠实验性牙周炎的影响。将 188 周大的雄性 C57BL/6 小鼠随机分为三组：对照组（Con）、牙周炎组（Peri）和 L-D-sEV 组。小鼠牙周炎模型是通过在上颌第二臼齿周围放置 5-0 丝线和牙龈脓毒性球菌（每只小鼠 1 ×107 CFUs）诱发的。结果在体外，服用 L-D-sEV 后，p-PDLCs 的 RANKL/OPG 比值、JNK 磷酸化和 P38 蛋白水平显著下降。此外，流式细胞术和 qRT-PCR 分析表明，L-D-sEV 可减少 p-PDLCs 的凋亡，下调凋亡相关基因 Caspase-3 和 BCL-2-Associated X 的表达，上调 B 细胞淋巴瘤-2 基因水平。在体内，L-D-sEV 能显著减少牙槽骨流失，抑制破骨细胞活性，诱导 M2 极化。组织学分析表明，L-D-sEV 组的 iNOS/CD206、RANKL/OPG、p-JNK/JNK 和 p-P38/P38 比率明显低于 Peri 组。

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引用次数: 0