Liu Liu, Tongfeng Fang, Cheng Miao, Xiangfen Li, Yanglin Zeng, Tianyi Wang, Yubin Cao, Dingming Huang, Dongzhe Song
Aim
The regenerative capacity of dental pulp relies on the odonto/osteogenic differentiation of dental pulp cells (DPCs), but dynamic microenvironmental changes hinder the process. Bone morphogenetic protein 9 (BMP9) promotes differentiation of DPCs towards an odonto/osteogenic lineage, forming dentinal-like tissue. However, the molecular mechanism underlying its action remains unclear. This study investigates the role of DLX6 antisense RNA 1 (DLX6-AS1) in odonto/osteogenic differentiation induced by BMP9.
Methodology
Custom RT2 profiler PCR array, quantitative Real-Time PCR (qRT-PCR) and western blots were used to investigate the expression pattern of DLX6-AS1 and its potential signal axis. Osteogenic ability was evaluated using alkaline phosphatase and alizarin red S staining. Interactions between lncRNA and miRNA, as well as miRNA and mRNA, were predicted through bioinformatic assays, which were subsequently validated via RNA immunoprecipitation and dual luciferase reporter assays. Student's t-test or one-way ANOVA with post hoc Tukey HSD tests were employed for data analysis, with a p-value of less than .05 considered statistically significant.
Results
DLX6-AS1 was upregulated upon BMP9 overexpression in DPCs, thereby promoting odonto/osteogenic differentiation. Additionally, miR-128-3p participated in BMP9-induced odonto/osteogenic differentiation by interacting with the downstream signal MAPK14. Modifying the expression of miR-128-3p and transfecting pcMAPK14/siMAPK14 had a rescue impact on odonto/osteogenic differentiation downstream of DLX6-AS1. Lastly, miR-128-3p directly interacted with both MAPK14 and DLX6-AS1.
Conclusions
DLX6-AS1 could regulate the odonto/osteogenic differentiation of DPCs under the control of BMP9 through the miR-128-3p/MAPK14 axis.
{"title":"DLX6-AS1 regulates odonto/osteogenic differentiation in dental pulp cells under the control of BMP9 via the miR-128-3p/MAPK14 axis: A laboratory investigation","authors":"Liu Liu, Tongfeng Fang, Cheng Miao, Xiangfen Li, Yanglin Zeng, Tianyi Wang, Yubin Cao, Dingming Huang, Dongzhe Song","doi":"10.1111/iej.14120","DOIUrl":"10.1111/iej.14120","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Aim</h3>\u0000 \u0000 <p>The regenerative capacity of dental pulp relies on the odonto/osteogenic differentiation of dental pulp cells (DPCs), but dynamic microenvironmental changes hinder the process. Bone morphogenetic protein 9 (BMP9) promotes differentiation of DPCs towards an odonto/osteogenic lineage, forming dentinal-like tissue. However, the molecular mechanism underlying its action remains unclear. This study investigates the role of DLX6 antisense RNA 1 (DLX6-AS1) in odonto/osteogenic differentiation induced by BMP9.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methodology</h3>\u0000 \u0000 <p>Custom RT<sup>2</sup> profiler PCR array, quantitative Real-Time PCR (qRT-PCR) and western blots were used to investigate the expression pattern of DLX6-AS1 and its potential signal axis. Osteogenic ability was evaluated using alkaline phosphatase and alizarin red S staining. Interactions between lncRNA and miRNA, as well as miRNA and mRNA, were predicted through bioinformatic assays, which were subsequently validated via RNA immunoprecipitation and dual luciferase reporter assays. Student's <i>t</i>-test or one-way ANOVA with post hoc Tukey HSD tests were employed for data analysis, with a <i>p</i>-value of less than .05 considered statistically significant.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>DLX6-AS1 was upregulated upon BMP9 overexpression in DPCs, thereby promoting odonto/osteogenic differentiation. Additionally, miR-128-3p participated in BMP9-induced odonto/osteogenic differentiation by interacting with the downstream signal MAPK14. Modifying the expression of miR-128-3p and transfecting pcMAPK14/siMAPK14 had a rescue impact on odonto/osteogenic differentiation downstream of DLX6-AS1. Lastly, miR-128-3p directly interacted with both MAPK14 and DLX6-AS1.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>DLX6-AS1 could regulate the odonto/osteogenic differentiation of DPCs under the control of BMP9 through the miR-128-3p/MAPK14 axis.</p>\u0000 </section>\u0000 </div>","PeriodicalId":13724,"journal":{"name":"International endodontic journal","volume":null,"pages":null},"PeriodicalIF":5.4,"publicationDate":"2024-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141554686","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
In this study, we investigated the systemic implications of chronic apical periodontitis (CAP). CAP may contribute to the nonalcoholic fatty liver disease (NAFLD) progression through the gut microbiota and its metabolites, which are related to the degree of fibrosis.
Methodology
Sixteen 7-week-old male apolipoprotein E knockout (apoE−/−) mice were randomly divided into two groups: the CAP and Con groups. A CAP model was established by sealing the first- and second-maxillary molars with bacterium-containing cotton balls. Apical lesions were evaluated by micro-CT. Histological evaluations of NAFLD were performed using second harmonic generation/two-photon excitation fluorescence (SHG/TPEF) assays. Additionally, we comprehensively analyzed the gut microbiota using 16S rRNA gene sequencing and explored metabolic profiles by liquid chromatography–mass spectrometry (LC–MS). Immunofluorescence analysis was used to examine the impact of CAP on tight junction proteins and mucin expression. Transcriptome assays have elucidated gene expression alterations in liver tissues.
Results
Micro-CT scans revealed an evident periapical bone loss in the CAP group, and the total collagen percentage was increased (Con, 0.0361 ± 0.00510%, CAP, 0.0589 ± 0.00731%, p < .05). 16S rRNA sequencing revealed reduced diversity and distinct taxonomic enrichment in the CAP group. Metabolomic assessments revealed that differentially enriched metabolites, including D-galactosamine, were enriched and that 16-hydroxyhexadecanoic acid and 3-methylindole were depleted in the CAP group. Immunofluorescence analyses revealed disruptions in tight junction proteins and mucin production, indicating intestinal barrier integrity disruption. Liver transcriptome analysis revealed upregulation of Lpin-1 expression in the CAP group.
Conclusion
This study provides comprehensive evidence of the systemic effects of CAP on liver fibrosis in NAFLD patients by elucidating alterations in the gut microbiota composition and metabolism.
目的：在这项研究中，我们探讨了慢性根尖牙周炎（CAP）的系统性影响。CAP可能通过肠道微生物群及其代谢产物导致非酒精性脂肪肝（NAFLD）的进展，而肠道微生物群及其代谢产物与纤维化程度有关：16只7周大的雄性载脂蛋白E基因敲除（apoE-/-）小鼠被随机分为两组：CAP组和Con组。通过用含菌棉球封闭第一和第二颌磨牙，建立 CAP 模型。通过显微 CT 对根尖病变进行评估。使用二次谐波发生/双光子激发荧光（SHG/TPEF）检测法对非酒精性脂肪肝进行组织学评估。此外，我们还利用 16S rRNA 基因测序对肠道微生物群进行了全面分析，并利用液相色谱-质谱法（LC-MS）对代谢谱进行了研究。免疫荧光分析用于研究 CAP 对紧密连接蛋白和粘蛋白表达的影响。转录组测定阐明了肝组织中基因表达的改变：结果：显微 CT 扫描显示，CAP 组根尖周骨质明显流失，总胶原蛋白百分比增加（Con，0.0361 ± 0.00510%；CAP，0.0589 ± 0.00731%，p 结论：该研究全面证实了 CAP 对全身骨质疏松症的影响：本研究通过阐明肠道微生物群组成和代谢的改变，为 CAP 对非酒精性脂肪肝患者肝纤维化的全身影响提供了全面的证据。
{"title":"Gut microbiota dysbiosis links chronic apical periodontitis to liver fibrosis in nonalcoholic fatty liver disease: Insights from a mouse model","authors":"Guowu Gan, Yufang Luo, Yu Zeng, Shihan Lin, Beibei Lu, Ren Zhang, Shuai Chen, Huaxiang Lei, Zhiyu Cai, Xiaojing Huang","doi":"10.1111/iej.14119","DOIUrl":"10.1111/iej.14119","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Aim</h3>\u0000 \u0000 <p>In this study, we investigated the systemic implications of chronic apical periodontitis (CAP). CAP may contribute to the nonalcoholic fatty liver disease (NAFLD) progression through the gut microbiota and its metabolites, which are related to the degree of fibrosis.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methodology</h3>\u0000 \u0000 <p>Sixteen 7-week-old male apolipoprotein E knockout (apoE<sup>−/−</sup>) mice were randomly divided into two groups: the CAP and Con groups. A CAP model was established by sealing the first- and second-maxillary molars with bacterium-containing cotton balls. Apical lesions were evaluated by micro-CT. Histological evaluations of NAFLD were performed using second harmonic generation/two-photon excitation fluorescence (SHG/TPEF) assays. Additionally, we comprehensively analyzed the gut microbiota using 16S rRNA gene sequencing and explored metabolic profiles by liquid chromatography–mass spectrometry (LC–MS). Immunofluorescence analysis was used to examine the impact of CAP on tight junction proteins and mucin expression. Transcriptome assays have elucidated gene expression alterations in liver tissues.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Micro-CT scans revealed an evident periapical bone loss in the CAP group, and the total collagen percentage was increased (Con, 0.0361 ± 0.00510%, CAP, 0.0589 ± 0.00731%, <i>p</i> < .05). 16S rRNA sequencing revealed reduced diversity and distinct taxonomic enrichment in the CAP group. Metabolomic assessments revealed that differentially enriched metabolites, including D-galactosamine, were enriched and that 16-hydroxyhexadecanoic acid and 3-methylindole were depleted in the CAP group. Immunofluorescence analyses revealed disruptions in tight junction proteins and mucin production, indicating intestinal barrier integrity disruption. Liver transcriptome analysis revealed upregulation of Lpin-1 expression in the CAP group.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>This study provides comprehensive evidence of the systemic effects of CAP on liver fibrosis in NAFLD patients by elucidating alterations in the gut microbiota composition and metabolism.</p>\u0000 </section>\u0000 </div>","PeriodicalId":13724,"journal":{"name":"International endodontic journal","volume":null,"pages":null},"PeriodicalIF":5.4,"publicationDate":"2024-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141491812","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
S. R. Patel, F. Jarad, E. Moawad, A. Boland, J. Greenhalgh, Maria Liu, Michelle Maden
<div> <section> <h3> Background</h3> <p>Analysis of the survival of root-filled posterior teeth and the associated prognostic tooth-related factors will enable clinicians to predict the outcome of root canal treatment.</p> </section> <section> <h3> Objectives</h3> <p>To investigate (i) the survival of root-filled posterior teeth and (ii) the tooth-related factors that may affect their survival.</p> </section> <section> <h3> Methods</h3> <p>Randomized controlled trials, comparative studies and observational studies assessing survival rates of root-filled posterior teeth with a minimum 4-year follow-up period were identified through an electronic search of the following databases up to January 2023: The Cochrane Central Register of Controlled Trials, Medline via PubMed, the Cochrane Database of Systematic Reviews, Embase, Web of Science and NIHR centre for reviews and dissemination. Two reviewers (SP and ML) independently selected the final studies based on pre-defined inclusion criteria. The Newcastle Ottawa Scale and the Cochrane Risk of Bias Tool for Randomized Trials were used to assess the risk of bias. Pooled weighted survival rates were analysed using a random effects meta-analysis model using DerSimonean and Laird methods. Descriptive analysis of studies describing any prognostic tooth-related factors was conducted.</p> </section> <section> <h3> Results</h3> <p>Of the 72 studies identified, data from 20 studies were included in the survival meta-analysis, and data from 13 of these studies were included in the descriptive analysis of tooth-related factors; 12 studies were retrospective, 7 were prospective, and one was a randomized control trial. The pooled survival rates at 4–7 years and 8–20 years of root-filled posterior teeth regardless of tooth type was 91% (95% CI, 0.85; 0.95) and 87% (95% CI, 0.77; 0.93), respectively. The prognostic tooth-related factors mentioned in the included studies were (i) remaining coronal tooth structure, (ii) ferrule, (iii) crown-to-root ratio (iv) tooth type and location (v) periodontal disease (vi) proximal contacts and (vii) cracks.</p> </section> <section> <h3> Conclusions</h3> <p>The meta-analysis suggests that root canal treatment has a high medium to long term survival outcome. The narrative summary identified 7 factors that affect tooth survival. However, there is a paucity of evidence, and more research is needed in this area.</p> </section> <section>
{"title":"The tooth survival of non-surgical root-filled posterior teeth and the associated prognostic tooth-related factors: A systematic review and meta-analysis","authors":"S. R. Patel, F. Jarad, E. Moawad, A. Boland, J. Greenhalgh, Maria Liu, Michelle Maden","doi":"10.1111/iej.14116","DOIUrl":"10.1111/iej.14116","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Analysis of the survival of root-filled posterior teeth and the associated prognostic tooth-related factors will enable clinicians to predict the outcome of root canal treatment.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Objectives</h3>\u0000 \u0000 <p>To investigate (i) the survival of root-filled posterior teeth and (ii) the tooth-related factors that may affect their survival.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Randomized controlled trials, comparative studies and observational studies assessing survival rates of root-filled posterior teeth with a minimum 4-year follow-up period were identified through an electronic search of the following databases up to January 2023: The Cochrane Central Register of Controlled Trials, Medline via PubMed, the Cochrane Database of Systematic Reviews, Embase, Web of Science and NIHR centre for reviews and dissemination. Two reviewers (SP and ML) independently selected the final studies based on pre-defined inclusion criteria. The Newcastle Ottawa Scale and the Cochrane Risk of Bias Tool for Randomized Trials were used to assess the risk of bias. Pooled weighted survival rates were analysed using a random effects meta-analysis model using DerSimonean and Laird methods. Descriptive analysis of studies describing any prognostic tooth-related factors was conducted.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Of the 72 studies identified, data from 20 studies were included in the survival meta-analysis, and data from 13 of these studies were included in the descriptive analysis of tooth-related factors; 12 studies were retrospective, 7 were prospective, and one was a randomized control trial. The pooled survival rates at 4–7 years and 8–20 years of root-filled posterior teeth regardless of tooth type was 91% (95% CI, 0.85; 0.95) and 87% (95% CI, 0.77; 0.93), respectively. The prognostic tooth-related factors mentioned in the included studies were (i) remaining coronal tooth structure, (ii) ferrule, (iii) crown-to-root ratio (iv) tooth type and location (v) periodontal disease (vi) proximal contacts and (vii) cracks.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>The meta-analysis suggests that root canal treatment has a high medium to long term survival outcome. The narrative summary identified 7 factors that affect tooth survival. However, there is a paucity of evidence, and more research is needed in this area.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 ","PeriodicalId":13724,"journal":{"name":"International endodontic journal","volume":null,"pages":null},"PeriodicalIF":5.4,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/iej.14116","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141467796","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Alexandre Henrique dos Reis-Prado, Caroline Andrade Maia, Gabriel Pereira Nunes, Lara Cancella de Arantes, Lucas Guimarães Abreu, Henry F. Duncan, Marco C. Bottino, Francine Benetti
<div> <section> <h3> Background</h3> <p>Bibliometric analysis is a critical indicator of the influence and relevance of scientific papers, whilst also highlighting key contributors and gaps in knowledge in a scientific field.</p> </section> <section> <h3> Objectives</h3> <p>To update and analyse the 100 most-cited papers in regenerative endodontics from 2019 to 2023.</p> </section> <section> <h3> Methods</h3> <p>A search of the most-cited recent papers focusing on regenerative endodontics using journals included in the category, ‘Dentistry, Oral Surgery & Medicine’, in the Clarivate Web of Science database from 2019 to 2023 was performed. Three researchers conducted the study selection and data extraction. Data extraction included publication title and year, authors, number and mean number of citations, institution, country and continent, study design, journal title, keywords and research topic. Citation counts were also collected in Google Scholar and Scopus databases. Graphical bibliometric networks were created using VOSviewer software.</p> </section> <section> <h3> Results</h3> <p>The number of citations of the 100 most-cited articles ranged from 6 to 85. Most were published in 2020 (<i>n</i> = 48), principally in the <i>Journal of Endodontics</i> (47%), followed by <i>International Endodontic Journal</i> (13%), <i>Journal of Dental Research</i> (6%) and <i>Dental Materials</i> (6%). Laboratory study was the most common study design amongst the included papers (<i>n</i> = 47), followed by narrative reviews (<i>n</i> = 17) and observational studies (<i>n</i> = 16). The most frequent first author on the top three most-cited papers was Hacer Aksel, whilst Adham A. Azim (<i>n</i> = 6; 89 citations) contributed most to the top 100 articles. The institution from which most articles originated was the University of Hong Kong (China) (<i>n</i> = 5; 81 citations), whereas the corresponding authors were predominantly from the United States of America (USA) (<i>n</i> = 31; 560 citations). The VOSviewer map of co-authorship demonstrated research collaborative clusters. ‘Regenerative endodontics’ and ‘stem-cells’ were the most employed keywords (37 and 36 occurrences respectively).</p> </section> <section> <h3> Discussion</h3> <p>The current study was designed not only to showcase the most influential papers in regenerative endodontics since 2019 but also to provide a better understanding of global research in this area over the last five years.</p>
{"title":"Top 100 most-cited scientific articles in regenerative endodontics 2019–2023: A bibliometric analysis","authors":"Alexandre Henrique dos Reis-Prado, Caroline Andrade Maia, Gabriel Pereira Nunes, Lara Cancella de Arantes, Lucas Guimarães Abreu, Henry F. Duncan, Marco C. Bottino, Francine Benetti","doi":"10.1111/iej.14117","DOIUrl":"10.1111/iej.14117","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Bibliometric analysis is a critical indicator of the influence and relevance of scientific papers, whilst also highlighting key contributors and gaps in knowledge in a scientific field.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Objectives</h3>\u0000 \u0000 <p>To update and analyse the 100 most-cited papers in regenerative endodontics from 2019 to 2023.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>A search of the most-cited recent papers focusing on regenerative endodontics using journals included in the category, ‘Dentistry, Oral Surgery & Medicine’, in the Clarivate Web of Science database from 2019 to 2023 was performed. Three researchers conducted the study selection and data extraction. Data extraction included publication title and year, authors, number and mean number of citations, institution, country and continent, study design, journal title, keywords and research topic. Citation counts were also collected in Google Scholar and Scopus databases. Graphical bibliometric networks were created using VOSviewer software.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The number of citations of the 100 most-cited articles ranged from 6 to 85. Most were published in 2020 (<i>n</i> = 48), principally in the <i>Journal of Endodontics</i> (47%), followed by <i>International Endodontic Journal</i> (13%), <i>Journal of Dental Research</i> (6%) and <i>Dental Materials</i> (6%). Laboratory study was the most common study design amongst the included papers (<i>n</i> = 47), followed by narrative reviews (<i>n</i> = 17) and observational studies (<i>n</i> = 16). The most frequent first author on the top three most-cited papers was Hacer Aksel, whilst Adham A. Azim (<i>n</i> = 6; 89 citations) contributed most to the top 100 articles. The institution from which most articles originated was the University of Hong Kong (China) (<i>n</i> = 5; 81 citations), whereas the corresponding authors were predominantly from the United States of America (USA) (<i>n</i> = 31; 560 citations). The VOSviewer map of co-authorship demonstrated research collaborative clusters. ‘Regenerative endodontics’ and ‘stem-cells’ were the most employed keywords (37 and 36 occurrences respectively).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Discussion</h3>\u0000 \u0000 <p>The current study was designed not only to showcase the most influential papers in regenerative endodontics since 2019 but also to provide a better understanding of global research in this area over the last five years.</p>\u0000 ","PeriodicalId":13724,"journal":{"name":"International endodontic journal","volume":null,"pages":null},"PeriodicalIF":5.4,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/iej.14117","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141467797","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Chiara Bernardini, Fausto Zamparini, Carlo Prati, Roberta Salaroli, Andrea Spinelli, Augusta Zannoni, Monica Forni, Maria Giovanna Gandolfi
Aim
The osteogenic potential of new premixed calcium-silicate-containing bioceramic sealers (Ca-Si sealers) was tested with porcine vascular wall-mesenchymal stem cells (pVW-MSCs).
Methodology
Two Ca-Si-containing sealers: Ceraseal (MetaBiomed, Cheong-si, South Korea) and AH Plus Bioceramic (Maruchi, Wonju-si, South Korea), and an epoxy resin sealer (AH Plus; Dentsply, Konstanz, Germany) as a control, were prepared according to the manufacturers' indications. All samples were allowed to set for 100% of their setting time in a sterile humid cabinet at 37°C and 95% relative humidity. pVW-MSC seeding efficiency and osteogenic differentiation were analysed as marker of gene/protein expression for up to 12 days. Mineralization assay and immunofluorescence staining were performed and evaluated over a period of 21 days. Statistical analyses were conducted using one-way analysis of variance (p < .05). Additional samples were prepared and stored under the same conditions and inspected using an environmental scanning electron microscope equipped with an energy dispersive X-ray spectroscopy system.
Results
Significantly higher cell seeding efficiency (p < .05) was observed for both Ca-Si sealers from day 8. pVW-MSCs showed a significant shift towards the osteogenic lineage only when seeded in contact with Ca-Si sealers. Gene expression of osteopontin was upregulated significantly. Collagen I and osteocalcin were clearly expressed by cells in contact with Ca-Si sealers. Mineralization granules were observed in Alizarin red assays and confocal laser scanning microscopy analysis of both Ca-Si sealers. No gene expression or granule mineralization were observed on the epoxy resin sealer.
Conclusions
Premixed Ca-Si sealers displayed a higher potential for osteogenic activity on pVW-MSCs. Epoxy resin sealer was unable to induce any osteogenic activity. The properties of both Ca-Si sealers suggest their potential as osteoinductive platforms for vascular MSCs in periapical bone.
{"title":"Osteoinductive and regenerative potential of premixed calcium-silicate bioceramic sealers on vascular wall mesenchymal stem cells","authors":"Chiara Bernardini, Fausto Zamparini, Carlo Prati, Roberta Salaroli, Andrea Spinelli, Augusta Zannoni, Monica Forni, Maria Giovanna Gandolfi","doi":"10.1111/iej.14098","DOIUrl":"10.1111/iej.14098","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Aim</h3>\u0000 \u0000 <p>The osteogenic potential of new premixed calcium-silicate-containing bioceramic sealers (Ca-Si sealers) was tested with porcine vascular wall-mesenchymal stem cells (pVW-MSCs).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methodology</h3>\u0000 \u0000 <p>Two Ca-Si-containing sealers: Ceraseal (MetaBiomed, Cheong-si, South Korea) and AH Plus Bioceramic (Maruchi, Wonju-si, South Korea), and an epoxy resin sealer (AH Plus; Dentsply, Konstanz, Germany) as a control, were prepared according to the manufacturers' indications. All samples were allowed to set for 100% of their setting time in a sterile humid cabinet at 37°C and 95% relative humidity. pVW-MSC seeding efficiency and osteogenic differentiation were analysed as marker of gene/protein expression for up to 12 days. Mineralization assay and immunofluorescence staining were performed and evaluated over a period of 21 days. Statistical analyses were conducted using one-way analysis of variance (<i>p</i> < .05). Additional samples were prepared and stored under the same conditions and inspected using an environmental scanning electron microscope equipped with an energy dispersive X-ray spectroscopy system.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Significantly higher cell seeding efficiency (<i>p</i> < .05) was observed for both Ca-Si sealers from day 8. pVW-MSCs showed a significant shift towards the osteogenic lineage only when seeded in contact with Ca-Si sealers. Gene expression of osteopontin was upregulated significantly. Collagen I and osteocalcin were clearly expressed by cells in contact with Ca-Si sealers. Mineralization granules were observed in Alizarin red assays and confocal laser scanning microscopy analysis of both Ca-Si sealers. No gene expression or granule mineralization were observed on the epoxy resin sealer.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>Premixed Ca-Si sealers displayed a higher potential for osteogenic activity on pVW-MSCs. Epoxy resin sealer was unable to induce any osteogenic activity. The properties of both Ca-Si sealers suggest their potential as osteoinductive platforms for vascular MSCs in periapical bone.</p>\u0000 </section>\u0000 </div>","PeriodicalId":13724,"journal":{"name":"International endodontic journal","volume":null,"pages":null},"PeriodicalIF":5.4,"publicationDate":"2024-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/iej.14098","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141467795","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Zhiwu Wu, Shaoying Duan, Mingming Li, Aopeng Zhang, Hui Yang, Jingjing Luo, Ran Cheng, Tao Hu
Aim
Autophagy is involved in human apical periodontitis (AP). However, it is not clear whether autophagy is protective or destructive in bone loss via the receptor activator of nuclear factor-κB ligand (RANKL)/RANK/osteoprotegerin (OPG) axis. This study aimed to investigate the involvement of autophagy via the RANKL/RANK/OPG axis during the development of AP in an experimental rat model.
Methodology
Twenty-four female Sprague–Dawley rats were divided into control, experimental AP (EAP) + saline, and EAP + 3-methyladenine (An autophagy inhibitor, 3-MA) groups. The control group did not receive any treatment. The EAP + saline group and the EAP + 3-MA group received intraperitoneal injections of saline and 3-MA, respectively, starting 1 week after the pulp was exposed. Specimens were collected for microcomputed tomography (micro-CT) scanning, histological processing, and immunostaining to examine the expression of light chain 3 beta (LC3B), RANK, RANKL, and OPG. Data were analysed using one-way analysis of variance (p < .05).
Results
Micro-CT showed greater bone loss in the EAP + 3-MA group than in the EAP + saline group, indicated by an elevated trabecular space (Tb.Sp) (p < .05). Inflammatory cell infiltration was observed in the EAP + saline and EAP + 3-MA groups. Compared with EAP + saline group, the EAP + 3-MA group showed weaker expression of LC3B (p < .01) and OPG (p < .05), more intense expression of RANK (p < .01) and RANKL (p < .01), and a higher RANKL/OPG ratio (p < .05).
Conclusion
Autophagy may exert a protective effect against AP by regulating the RANKL/RANK/OPG axis, thereby inhibiting excessive bone loss.
{"title":"Autophagy regulates bone loss via the RANKL/RANK/OPG axis in an experimental rat apical periodontitis model","authors":"Zhiwu Wu, Shaoying Duan, Mingming Li, Aopeng Zhang, Hui Yang, Jingjing Luo, Ran Cheng, Tao Hu","doi":"10.1111/iej.14103","DOIUrl":"10.1111/iej.14103","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Aim</h3>\u0000 \u0000 <p>Autophagy is involved in human apical periodontitis (AP). However, it is not clear whether autophagy is protective or destructive in bone loss via the receptor activator of nuclear factor-κB ligand (RANKL)/RANK/osteoprotegerin (OPG) axis. This study aimed to investigate the involvement of autophagy via the RANKL/RANK/OPG axis during the development of AP in an experimental rat model.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methodology</h3>\u0000 \u0000 <p>Twenty-four female Sprague–Dawley rats were divided into control, experimental AP (EAP) + saline, and EAP + 3-methyladenine (An autophagy inhibitor, 3-MA) groups. The control group did not receive any treatment. The EAP + saline group and the EAP + 3-MA group received intraperitoneal injections of saline and 3-MA, respectively, starting 1 week after the pulp was exposed. Specimens were collected for microcomputed tomography (micro-CT) scanning, histological processing, and immunostaining to examine the expression of light chain 3 beta (LC3B), RANK, RANKL, and OPG. Data were analysed using one-way analysis of variance (<i>p</i> < .05).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Micro-CT showed greater bone loss in the EAP + 3-MA group than in the EAP + saline group, indicated by an elevated trabecular space (Tb.Sp) (<i>p</i> < .05). Inflammatory cell infiltration was observed in the EAP + saline and EAP + 3-MA groups. Compared with EAP + saline group, the EAP + 3-MA group showed weaker expression of LC3B (<i>p</i> < .01) and OPG (<i>p</i> < .05), more intense expression of RANK (<i>p</i> < .01) and RANKL (<i>p</i> < .01), and a higher RANKL/OPG ratio (<i>p</i> < .05).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>Autophagy may exert a protective effect against AP by regulating the RANKL/RANK/OPG axis, thereby inhibiting excessive bone loss.</p>\u0000 </section>\u0000 </div>","PeriodicalId":13724,"journal":{"name":"International endodontic journal","volume":null,"pages":null},"PeriodicalIF":5.4,"publicationDate":"2024-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141456583","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Si Yu, Xue-Mei Liu, Yao Liu, Lu Tang, Shuang Lei, Chang Geng, Zhengwei Yuan, Xu Chen
Aim
This study investigated the effects of the inflammatory microenvironment of moderate pulpitis on biological properties of human dental pulp stem cells (DPSCs) and further explored the mechanism involved in osteo-/odontogenic induction of the inflammatory microenvironment.
Methodology
Healthy DPSCs (hDPSCs) and inflammatory DPSCs (iDPSCs) were isolated from human-impacted third molars free of caries and clinically diagnosed with moderate pulpitis, respectively. Healthy DPSCs were treated with lipopolysaccharides (LPS) to mimic iDPSCs in vitro. The surface markers expressed on hDPSCs and iDPSCs were detected by flow cytometry. A CCK-8 assay was performed to determine cell proliferation. Flow cytometric analysis was used to evaluate cell apoptosis. The osteo-/odontogenic differentiation of DPSCs was evaluated by western blot, alkaline phosphatase staining, and Alizarin Red S staining. The functions of the genes of differentially expressed mRNAs of hDPSCs and iDPSCs were analysed using gene set enrichment analysis. Transmission electron microscopy and western blot were used to evaluate the autophagy changes of LPS-treated DPSCs.
Results
Compared with hDPSCs, iDPSCs showed no significant difference in proliferative capacity but had stronger osteo-/odontogenic potential. In addition, the mRNAs differentially expressed between iDPSCs and hDPSCs were considerably enriched in autophagosome formation and assembly-related molecules. In vitro mechanism studies further found that low concentrations of LPS could upregulate DPSC autophagy-related protein expression and autophagosome formation and promote its odontogenic/osteogenic differentiation, whereas the inhibition of DPSC autophagy led to the weakening of the odontogenic/osteogenic differentiation induced by LPS.
Conclusions
This explorative study showed that DPSCs isolated from teeth with moderate pulpitis possessed higher osteo-/odontogenic differentiation capacity, and the mechanism involved was related to the inflammatory microenvironment-mediated autophagy of DPSCs. This helps to better understand the repair potential of inflamed dental pulp and provides the biological basis for pulp preservation and hard tissue formation in minimally invasive endodontics.
{"title":"Inflammatory microenvironment of moderate pulpitis enhances the osteo-/odontogenic potential of dental pulp stem cells by autophagy","authors":"Si Yu, Xue-Mei Liu, Yao Liu, Lu Tang, Shuang Lei, Chang Geng, Zhengwei Yuan, Xu Chen","doi":"10.1111/iej.14108","DOIUrl":"10.1111/iej.14108","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Aim</h3>\u0000 \u0000 <p>This study investigated the effects of the inflammatory microenvironment of moderate pulpitis on biological properties of human dental pulp stem cells (DPSCs) and further explored the mechanism involved in osteo-/odontogenic induction of the inflammatory microenvironment.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methodology</h3>\u0000 \u0000 <p>Healthy DPSCs (hDPSCs) and inflammatory DPSCs (iDPSCs) were isolated from human-impacted third molars free of caries and clinically diagnosed with moderate pulpitis, respectively. Healthy DPSCs were treated with lipopolysaccharides (LPS) to mimic iDPSCs in vitro. The surface markers expressed on hDPSCs and iDPSCs were detected by flow cytometry. A CCK-8 assay was performed to determine cell proliferation. Flow cytometric analysis was used to evaluate cell apoptosis. The osteo-/odontogenic differentiation of DPSCs was evaluated by western blot, alkaline phosphatase staining, and Alizarin Red S staining. The functions of the genes of differentially expressed mRNAs of hDPSCs and iDPSCs were analysed using gene set enrichment analysis. Transmission electron microscopy and western blot were used to evaluate the autophagy changes of LPS-treated DPSCs.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Compared with hDPSCs, iDPSCs showed no significant difference in proliferative capacity but had stronger osteo-/odontogenic potential. In addition, the mRNAs differentially expressed between iDPSCs and hDPSCs were considerably enriched in autophagosome formation and assembly-related molecules. In vitro mechanism studies further found that low concentrations of LPS could upregulate DPSC autophagy-related protein expression and autophagosome formation and promote its odontogenic/osteogenic differentiation, whereas the inhibition of DPSC autophagy led to the weakening of the odontogenic/osteogenic differentiation induced by LPS.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>This explorative study showed that DPSCs isolated from teeth with moderate pulpitis possessed higher osteo-/odontogenic differentiation capacity, and the mechanism involved was related to the inflammatory microenvironment-mediated autophagy of DPSCs. This helps to better understand the repair potential of inflamed dental pulp and provides the biological basis for pulp preservation and hard tissue formation in minimally invasive endodontics.</p>\u0000 </section>\u0000 </div>","PeriodicalId":13724,"journal":{"name":"International endodontic journal","volume":null,"pages":null},"PeriodicalIF":5.4,"publicationDate":"2024-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141507285","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sally McCarthy, Kishor Gulabivala, Geoffrey St. George, Simon Harvey, Yuan-Ling Ng
Aim
To explore self-reported dentofacial trauma and their potential endodontic sequelae in boxers using a questionnaire, followed by clinical and radiographic assessment to (1) compare the nature and number of self-reported dentofacial injuries with physical evidence of injury sequelae; and (2) investigate potential risk factors influencing dentofacial trauma and their endodontic sequelae.
Methodology
A focus group validated questionnaire was completed by 176 boxers recruited from 16 London boxing clubs; 61 boxers from this cohort then attended a London dental hospital, for a clinical and radiographic assessment. Data from the questionnaire and clinical assessments were then collated and analysed using Chi-squared or t-tests.
Results
Questionnaire data revealed 87.5% of boxers reported a history of dentofacial trauma during boxing activity. The clinical and radiographic assessment detected evidence of dentofacial trauma in 91.8% of boxers and dental injury or endodontic-related injury sequelae in 68.9% of boxers. There was a significant association between dentofacial trauma and boxers who did not participate in weekly neck weight sessions (p < .001), and there was a significant association between trauma-related endodontic sequelae and: boxer age (p = .01); competitions per month (p = .002); and defensive skill (p = .007).
Conclusions
A majority of the cohort had suffered dentofacial injuries and endodontic sequelae. The questionnaire data under-reported musculoskeletal injuries and endodontic sequelae, suggesting that some hard-tissue injuries following repetitive dentofacial trauma may have a subclinical presentation. Injury risk may be related to increased boxer age, defensive skills, frequency of participation in competitions, and frequency of neck weight sessions per week.
{"title":"Endodontic sequelae associated with repetitive impacts to the dentofacial region during boxing activities","authors":"Sally McCarthy, Kishor Gulabivala, Geoffrey St. George, Simon Harvey, Yuan-Ling Ng","doi":"10.1111/iej.14111","DOIUrl":"10.1111/iej.14111","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Aim</h3>\u0000 \u0000 <p>To explore <i>self-reported</i> dentofacial trauma and their potential endodontic sequelae in boxers using a questionnaire, followed by clinical and radiographic assessment to (1) compare the nature and number of <i>self-reported</i> dentofacial injuries with physical evidence of injury sequelae; and (2) investigate potential risk factors influencing dentofacial trauma and their endodontic sequelae.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methodology</h3>\u0000 \u0000 <p>A focus group validated questionnaire was completed by 176 boxers recruited from 16 London boxing clubs; 61 boxers from this cohort then attended a London dental hospital, for a clinical and radiographic assessment. Data from the questionnaire and clinical assessments were then collated and analysed using Chi-squared or <i>t</i>-tests.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Questionnaire data revealed 87.5% of boxers reported a history of dentofacial trauma during boxing activity. The clinical and radiographic assessment detected evidence of dentofacial trauma in 91.8% of boxers and dental injury or endodontic-related injury sequelae in 68.9% of boxers. There was a significant association between dentofacial trauma and boxers who did not participate in weekly neck weight sessions (<i>p</i> < .001), and there was a significant association between trauma-related endodontic sequelae and: boxer age (<i>p</i> = .01); competitions per month (<i>p</i> = .002); and defensive skill (<i>p</i> = .007).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>A majority of the cohort had suffered dentofacial injuries and endodontic sequelae. The questionnaire data under-reported musculoskeletal injuries and endodontic sequelae, suggesting that some hard-tissue injuries following repetitive dentofacial trauma may have a subclinical presentation. Injury risk may be related to increased boxer age, defensive skills, frequency of participation in competitions, and frequency of neck weight sessions per week.</p>\u0000 </section>\u0000 </div>","PeriodicalId":13724,"journal":{"name":"International endodontic journal","volume":null,"pages":null},"PeriodicalIF":5.4,"publicationDate":"2024-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/iej.14111","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141507284","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Tanujaa Suriyanarayanan, Lye Siang Lee, Sharon Hong Yu Han, Jianhong Ching, Chaminda J. Seneviratne
Aim
(i) To characterize Enterococcus faecalis biofilm formation pathways by semi-targeted metabolomics and targeted nitrogen panel analysis of strong (Ef63) and weak (Ef 64) biofilm forming E. faecalis clinical isolates and (ii) to validate the identified metabolic markers using targeted inhibitors.
Methodology
Previous proteomics profiling of E. faecalis clinical isolates with strong and weak biofilm formation revealed that differences in metabolic activity levels of small molecule, nucleotide and nitrogen compound metabolic processes and biosynthetic pathways, cofactor metabolic process, cellular amino acid and derivative metabolic process and lyase activity were associated with differences in biofilm formation. Hence, semi-targeted analysis of Ef 63, Ef 64 and ATC control strain Ef 29212 was performed by selecting metabolites that were part of both the previously identified pathways and a curated library with confirmed physical and chemical identity, followed by confirmatory targeted nitrogen panel analysis. Significantly regulated metabolites (p < .05) were selected based on fold change cut-offs of 1.2 and 0.8 for upregulation and downregulation, respectively, and subjected to pathway enrichment analysis. The identified metabolites and pathways were validated by minimum biofilm inhibitory concentration (MBIC) and colony forming unit (CFU) assays with targeted inhibitors.
Results
Metabolomics analysis showed upregulation of betaine, hypoxanthine, glycerophosphorylcholine, tyrosine, inosine, allantoin and citrulline in Ef 63 w.r.t Ef 64 and Ef 29212, and thesemetabolites mapped to purinemetabolism, urea cycle and aspartate metabolism pathways. MBIC and CFU assays using compounds against selected metabolites and metabolic pathways, namely glutathione against hypoxanthine and hydroxylamine against aspartate metabolism showed inhibitory effects against E. faecalis biofilm formation.
Conclusions
The study demonstrated the importance of oxidative stress inducers such as hypoxanthine and aspartate metabolism pathway in E. faecalis biofilm formation. Targeted therapeutics against these metabolic markers can reduce the healthcare burden associated with E. faecalis infections.
目的：(i) 通过对强（Ef63）和弱（Ef64）生物膜形成的粪肠球菌临床分离株进行半靶向代谢组学和靶向氮面板分析，确定粪肠球菌生物膜形成途径的特征；(ii) 使用靶向抑制剂验证已确定的代谢标记物：之前对具有强生物膜形成能力和弱生物膜形成能力的粪大肠杆菌临床分离株进行的蛋白质组学分析表明，小分子、核苷酸和氮化合物代谢过程和生物合成途径、辅助因子代谢过程、细胞氨基酸和衍生物代谢过程以及裂解酶活性水平的差异与生物膜形成的差异有关。因此，对 Ef 63、Ef 64 和 ATC 对照菌株 Ef 29212 进行了半靶向分析，选择了既属于先前确定的途径又属于已确认物理和化学特性的策划文库的代谢物，然后进行了确认性靶向氮小组分析。受显著调控的代谢物（p 结果：代谢组学分析表明，与 Ef 64 和 Ef 29212 相比，Ef 63 中的甜菜碱、次黄嘌呤、甘油磷酸胆碱、酪氨酸、肌苷、尿囊素和瓜氨酸出现上调，这些代谢物与嘌呤代谢、尿素循环和天冬氨酸代谢途径相关。利用针对选定代谢物和代谢途径的化合物（即针对次黄嘌呤代谢的谷胱甘肽和针对天冬氨酸代谢的羟胺）进行的 MBIC 和 CFU 检测显示，这些化合物对粪肠杆菌生物膜的形成具有抑制作用：该研究证明了氧化应激诱导剂（如次黄嘌呤和天门冬氨酸代谢途径）在粪大肠杆菌生物膜形成中的重要性。针对这些代谢标记物的靶向疗法可以减轻与粪大肠杆菌感染相关的医疗负担。
{"title":"Targeted metabolomics analysis approach to unravel the biofilm formation pathways of Enterococcus faecalis clinical isolates","authors":"Tanujaa Suriyanarayanan, Lye Siang Lee, Sharon Hong Yu Han, Jianhong Ching, Chaminda J. Seneviratne","doi":"10.1111/iej.14110","DOIUrl":"10.1111/iej.14110","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Aim</h3>\u0000 \u0000 <p>(i) To characterize <i>Enterococcus faecalis</i> biofilm formation pathways by semi-targeted metabolomics and targeted nitrogen panel analysis of strong (Ef63) and weak (Ef 64) biofilm forming <i>E. faecalis</i> clinical isolates and (ii) to validate the identified metabolic markers using targeted inhibitors.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methodology</h3>\u0000 \u0000 <p>Previous proteomics profiling of <i>E. faecalis</i> clinical isolates with strong and weak biofilm formation revealed that differences in metabolic activity levels of small molecule, nucleotide and nitrogen compound metabolic processes and biosynthetic pathways, cofactor metabolic process, cellular amino acid and derivative metabolic process and lyase activity were associated with differences in biofilm formation. Hence, semi-targeted analysis of Ef 63, Ef 64 and ATC control strain Ef 29212 was performed by selecting metabolites that were part of both the previously identified pathways and a curated library with confirmed physical and chemical identity, followed by confirmatory targeted nitrogen panel analysis. Significantly regulated metabolites (<i>p</i> < .05) were selected based on fold change cut-offs of 1.2 and 0.8 for upregulation and downregulation, respectively, and subjected to pathway enrichment analysis. The identified metabolites and pathways were validated by minimum biofilm inhibitory concentration (MBIC) and colony forming unit (CFU) assays with targeted inhibitors.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Metabolomics analysis showed upregulation of betaine, hypoxanthine, glycerophosphorylcholine, tyrosine, inosine, allantoin and citrulline in Ef 63 w.r.t Ef 64 and Ef 29212, and thesemetabolites mapped to purinemetabolism, urea cycle and aspartate metabolism pathways. MBIC and CFU assays using compounds against selected metabolites and metabolic pathways, namely glutathione against hypoxanthine and hydroxylamine against aspartate metabolism showed inhibitory effects against <i>E. faecalis</i> biofilm formation.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>The study demonstrated the importance of oxidative stress inducers such as hypoxanthine and aspartate metabolism pathway in <i>E. faecalis</i> biofilm formation. Targeted therapeutics against these metabolic markers can reduce the healthcare burden associated with <i>E. faecalis</i> infections.</p>\u0000 </section>\u0000 </div>","PeriodicalId":13724,"journal":{"name":"International endodontic journal","volume":null,"pages":null},"PeriodicalIF":5.4,"publicationDate":"2024-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141418841","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
<div> <section> <h3> Background</h3> <p>The dental pulp's environment is essential for the regulation of mesenchymal stem cells' homeostasis and thus, it is of great importance to evaluate the materials used in regenerative procedures.</p> </section> <section> <h3> Aim</h3> <p>To assess in vitro (i) the effect of chitosan nanoparticles, 0.2% chitosan irrigation solution, Dual Rinse®, 17% EDTA, 10% citric acid and 2.5% NaOCl on DSCS viability; (ii) the effect of different concentrations of TGF-β1 on DCSC proliferation; and (iii) whether treatment with TGF-β1 following exposure to the different irrigation solutions could compensate for their negative effects.</p> </section> <section> <h3> Methodology</h3> <p>(i) DSCS were treated with three dilutions (1:10, 1:100 and 1:1000) of the six irrigation solutions prepared in DMEM for 10 and 60 min to assess the effect on viability. (ii) The effect of different concentrations (0, 1, 5 and 10 ng/mL) of TGF-β1 on DCSC proliferation was assessed at 1, 3 and 7 days. (iii) The proliferative effect of TGF-β1 following 10-min exposure to 1:10 dilution of each irrigation solution was also tested. We used MTT assay to assess viability and proliferation. We performed statistical analysis using Prism software.</p> </section> <section> <h3> Results</h3> <p>(i) The different endodontic irrigation solutions tested showed a significant effect on cell viability (<i>p</i> ≤ .0001). Significant interactions between the endodontic irrigation solutions and their dilutions were also found for all parameters (<i>p</i> ≤ .0001). Chitosan nanoparticles and 0.2% chitosan irrigation solution were the least cytotoxic to DSCS whilst 2.5% NaOCl was the most cytotoxic followed by 17% EDTA. (ii) TGF-β1 at concentrations of 1 and 5 ng/mL resulted in significantly higher proliferation compared to the control group. (iii) Exposure to 17% EDTA or 2.5% NaOCl for 10 min was sufficient to make DSCS cells refractory to the proliferative effects of TGF-β1. DSCS groups treated with TGF-β1 following exposure to chitosan nanoparticles, 0.2% chitosan irrigation solution, Dual Rinse® and 10% CA demonstrated significantly higher proliferation compared to non-TGF-β1-treated groups (<i>p</i> ≤ .0001, <i>p</i> ≤ .0001, <i>p</i> ≤ .0001 and <i>p</i> = .01 respectively).</p> </section> <section> <h3> Conclusions</h3> <p>The current study offers data that can be implemented to improve the outcome of regenerative endodontic procedures by using less toxic irrig
{"title":"Chitosan-based endodontic irrigation solutions and TGF-β1 treatment: Creating the most favourable environment for the survival and proliferation of stem cells of the apical papilla in vitro","authors":"Roumaissa Belkadi, Diana Sanz-Serrano, Francesc Ventura, Montse Mercade","doi":"10.1111/iej.14112","DOIUrl":"10.1111/iej.14112","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>The dental pulp's environment is essential for the regulation of mesenchymal stem cells' homeostasis and thus, it is of great importance to evaluate the materials used in regenerative procedures.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Aim</h3>\u0000 \u0000 <p>To assess in vitro (i) the effect of chitosan nanoparticles, 0.2% chitosan irrigation solution, Dual Rinse®, 17% EDTA, 10% citric acid and 2.5% NaOCl on DSCS viability; (ii) the effect of different concentrations of TGF-β1 on DCSC proliferation; and (iii) whether treatment with TGF-β1 following exposure to the different irrigation solutions could compensate for their negative effects.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methodology</h3>\u0000 \u0000 <p>(i) DSCS were treated with three dilutions (1:10, 1:100 and 1:1000) of the six irrigation solutions prepared in DMEM for 10 and 60 min to assess the effect on viability. (ii) The effect of different concentrations (0, 1, 5 and 10 ng/mL) of TGF-β1 on DCSC proliferation was assessed at 1, 3 and 7 days. (iii) The proliferative effect of TGF-β1 following 10-min exposure to 1:10 dilution of each irrigation solution was also tested. We used MTT assay to assess viability and proliferation. We performed statistical analysis using Prism software.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>(i) The different endodontic irrigation solutions tested showed a significant effect on cell viability (<i>p</i> ≤ .0001). Significant interactions between the endodontic irrigation solutions and their dilutions were also found for all parameters (<i>p</i> ≤ .0001). Chitosan nanoparticles and 0.2% chitosan irrigation solution were the least cytotoxic to DSCS whilst 2.5% NaOCl was the most cytotoxic followed by 17% EDTA. (ii) TGF-β1 at concentrations of 1 and 5 ng/mL resulted in significantly higher proliferation compared to the control group. (iii) Exposure to 17% EDTA or 2.5% NaOCl for 10 min was sufficient to make DSCS cells refractory to the proliferative effects of TGF-β1. DSCS groups treated with TGF-β1 following exposure to chitosan nanoparticles, 0.2% chitosan irrigation solution, Dual Rinse® and 10% CA demonstrated significantly higher proliferation compared to non-TGF-β1-treated groups (<i>p</i> ≤ .0001, <i>p</i> ≤ .0001, <i>p</i> ≤ .0001 and <i>p</i> = .01 respectively).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>The current study offers data that can be implemented to improve the outcome of regenerative endodontic procedures by using less toxic irrig","PeriodicalId":13724,"journal":{"name":"International endodontic journal","volume":null,"pages":null},"PeriodicalIF":5.4,"publicationDate":"2024-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/iej.14112","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141418840","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号