Pub Date : 2024-06-01DOI: 10.1016/j.job.2024.03.002
Tao Su , Yi-hui Chen , Kan-kui Wu , Xiao-hong Xu
Objectives
To elucidate the association between the anticancer activities of piperlongumine (PL) and its potential target, transient receptor potential melastatin 7 channel (TRPM7), in oral squamous cell carcinoma (OSCC).
Methods
The expression levels and electrical characteristics of TRPM7 as well as cell viability in response to various PL treatments were investigated in the OSCC cell line Cal27.
Results
PL treatment resulted in a concentration- and time-dependent reduction in TRPM7 mRNA and protein expression in Cal27 cells. Furthermore, PL treatment inhibited TRPM7-like rectifying currents in Cal27 cells; however, this inhibition was less effective than that of the TRPM7 antagonist waixenicin A. Rapid perfusion and washout experiments revealed an immediate inhibitory effect of PL on TRPM7-like currents. The antagonistic effect of PL occurred within 1 min and was not completely reversed following washout. Notably, the extracellular Ca2+ concentration still influenced PL-induced changes in the TRPM7-like current, indicating that PL can directly but gently antagonize the TRPM7 channel. Functional changes in TRPM7 correlated with the observed antiproliferative and cytotoxic effects of PL in Cal27 cells.
Conclusions
These findings suggest that PL exhibits potent inhibitory effects on TRPM7 and exerts its anti-cancer effects by downregulating TRPM7 expression and antagonizing channel currents.
{"title":"Anti-cancer agent piperlongumine is an inhibitor of transient receptor potential melastatin 7 channel in oral squamous cell carcinoma","authors":"Tao Su , Yi-hui Chen , Kan-kui Wu , Xiao-hong Xu","doi":"10.1016/j.job.2024.03.002","DOIUrl":"10.1016/j.job.2024.03.002","url":null,"abstract":"<div><h3>Objectives</h3><p>To elucidate the association between the anticancer activities of piperlongumine (PL) and its potential target, transient receptor potential melastatin 7 channel (TRPM7), in oral squamous cell carcinoma (OSCC).</p></div><div><h3>Methods</h3><p>The expression levels and electrical characteristics of TRPM7 as well as cell viability in response to various PL treatments were investigated in the OSCC cell line Cal27.</p></div><div><h3>Results</h3><p>PL treatment resulted in a concentration- and time-dependent reduction in <em>TRPM7</em> mRNA and protein expression in Cal27 cells. Furthermore, PL treatment inhibited TRPM7-like rectifying currents in Cal27 cells; however, this inhibition was less effective than that of the TRPM7 antagonist waixenicin A. Rapid perfusion and washout experiments revealed an immediate inhibitory effect of PL on TRPM7-like currents. The antagonistic effect of PL occurred within 1 min and was not completely reversed following washout. Notably, the extracellular Ca<sup>2+</sup> concentration still influenced PL-induced changes in the TRPM7-like current, indicating that PL can directly but gently antagonize the TRPM7 channel. Functional changes in TRPM7 correlated with the observed antiproliferative and cytotoxic effects of PL in Cal27 cells.</p></div><div><h3>Conclusions</h3><p>These findings suggest that PL exhibits potent inhibitory effects on TRPM7 and exerts its anti-cancer effects by downregulating TRPM7 expression and antagonizing channel currents.</p></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140060723","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-06-01DOI: 10.1016/j.job.2024.04.006
Boang Liu , Chiho Mashimo , Takayuki Nambu , Hugo Maruyama , Toshinori Okinaga
Objectives
Rothia spp. are emerging as significant bacteria associated with oral health, with Rothia dentocariosa being one of the most prevalent species. However, there is a lack of studies examining these properties at the genetic level. This study aimed to establish a genetic modification platform for R. dentocariosa.
Methods
Rothia spp. were isolated from saliva samples collected from healthy volunteers. Subsequently, R. dentocariosa strains were identified through colony morphology, species-specific polymerase chain reaction (PCR), and 16S ribosomal RNA gene sequencing. The identified strains were then transformed with plasmid pJRD215, and the most efficient strain was selected. Transposon insertion mutagenesis was performed to investigate the possibility of genetic modifications.
Results
A strain demonstrating high transforming ability, designated as R. dentocariosa LX16, was identified. This strain underwent transposon insertion mutagenesis and was screened for 5-fluoroorotic acid-resistant transposants. The insertion sites were confirmed using arbitrary primed PCR, gene-specific PCR, and Sanger sequencing.
Conclusion
This study marks the first successful genetic modification of R. dentocariosa. Investigating R. dentocariosa at the genetic level can provide insights into its role within the oral microbiome.
目的齿槽菌属正在成为与口腔健康相关的重要细菌，其中齿槽菌是最普遍的种类之一。然而，目前还缺乏从基因层面研究这些特性的研究。本研究的目的是为牙关紧闭杆菌建立一个基因改造平台。方法从健康志愿者的唾液样本中分离出牙关紧闭杆菌。随后，通过菌落形态学、物种特异性聚合酶链式反应（PCR）和 16S 核糖体 RNA 基因测序鉴定出牙关紧闭杆菌菌株。然后用质粒 pJRD215 对鉴定出的菌株进行转化，选出效率最高的菌株。结果 发现了一株转化能力很强的菌株，命名为 R. dentocariosa LX16。对该菌株进行了转座子插入诱变，并筛选出了抗 5-氟乳清酸的转座子。使用任意引物 PCR、基因特异性 PCR 和 Sanger 测序对插入位点进行了确认。在基因水平上研究 R. dentocariosa 可以深入了解其在口腔微生物组中的作用。
{"title":"Transposon insertion in Rothia dentocariosa","authors":"Boang Liu , Chiho Mashimo , Takayuki Nambu , Hugo Maruyama , Toshinori Okinaga","doi":"10.1016/j.job.2024.04.006","DOIUrl":"10.1016/j.job.2024.04.006","url":null,"abstract":"<div><h3>Objectives</h3><p><em>Rothia</em> spp. are emerging as significant bacteria associated with oral health, with <em>Rothia dentocariosa</em> being one of the most prevalent species. However, there is a lack of studies examining these properties at the genetic level. This study aimed to establish a genetic modification platform for <em>R. dentocariosa.</em></p></div><div><h3>Methods</h3><p><em>Rothia</em> spp. were isolated from saliva samples collected from healthy volunteers. Subsequently, <em>R. dentocariosa</em> strains were identified through colony morphology, species-specific polymerase chain reaction (PCR), and 16S ribosomal RNA gene sequencing. The identified strains were then transformed with plasmid pJRD215, and the most efficient strain was selected. Transposon insertion mutagenesis was performed to investigate the possibility of genetic modifications.</p></div><div><h3>Results</h3><p>A strain demonstrating high transforming ability, designated as <em>R. dentocariosa</em> LX16, was identified. This strain underwent transposon insertion mutagenesis and was screened for 5-fluoroorotic acid-resistant transposants. The insertion sites were confirmed using arbitrary primed PCR, gene-specific PCR, and Sanger sequencing.</p></div><div><h3>Conclusion</h3><p>This study marks the first successful genetic modification of <em>R. dentocariosa</em>. Investigating <em>R. dentocariosa</em> at the genetic level can provide insights into its role within the oral microbiome.</p></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140785624","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-06-01DOI: 10.1016/j.job.2024.03.005
Masae Furukawa , Hirobumi Tada , Resmi Raju , Jingshu Wang , Haruna Yokoi , Mitsuyoshi Yamada , Yosuke Shikama , Takashi Saito , Takaomi C. Saido , Kenji Matsushita
Objectives
Many patients with Alzheimer's disease (AD) experience behavioral and psychological symptoms of dementia (BPSD), which significantly affect their quality of life. It is known that 5-Hydroxytryptamine (5-HT) plays a crucial role in the development of BPSD. While the relationship between tooth loss and AD symptoms has been acknowledged, the aspect of aggression has not been focused on until now. Despite the established importance of 5-HT in BPSD, how tooth loss is related to the exacerbation of AD symptoms, especially in terms of aggression, remains largely unexplored. Although nutritional status is known to influence the progression of dementia, the specific effect of tooth loss on peripheral symptoms, notably aggression, is not well understood.
Methods
In our study, we conducted maxillary molar extractions in aged C57BL/6J and AppNL-G-F mice and observed their condition over a 3-month period. During this time, we documented significant behavioral and genetic differences between mice in the control groups and mice that underwent tooth extraction. Notably, mice that underwent tooth extraction exhibited a considerable decline in cognitive function and increased in aggression 3 months after tooth extraction compared with the control groups (C57BL/6J and AppNL-G-Fmice).
Results
Our findings suggest that molar loss may lead to reduced 5-HT levels in the hippocampus, possibly mediated by the trigeminal nerve, contributing to the development of aggression and BPSD in AD.
Conclusion
This study sheds light on the intricate relationships between oral health, 5-HT, and AD symptoms, offering valuable insights into potential therapeutic avenues for managing BPSD in patients with dementia.
{"title":"Effects of tooth loss on behavioral and psychological symptoms of dementia in app knock-in mice","authors":"Masae Furukawa , Hirobumi Tada , Resmi Raju , Jingshu Wang , Haruna Yokoi , Mitsuyoshi Yamada , Yosuke Shikama , Takashi Saito , Takaomi C. Saido , Kenji Matsushita","doi":"10.1016/j.job.2024.03.005","DOIUrl":"10.1016/j.job.2024.03.005","url":null,"abstract":"<div><h3>Objectives</h3><p>Many patients with Alzheimer's disease (AD) experience behavioral and psychological symptoms of dementia (BPSD), which significantly affect their quality of life. It is known that 5-Hydroxytryptamine (5-HT) plays a crucial role in the development of BPSD. While the relationship between tooth loss and AD symptoms has been acknowledged, the aspect of aggression has not been focused on until now. Despite the established importance of 5-HT in BPSD, how tooth loss is related to the exacerbation of AD symptoms, especially in terms of aggression, remains largely unexplored. Although nutritional status is known to influence the progression of dementia, the specific effect of tooth loss on peripheral symptoms, notably aggression, is not well understood.</p></div><div><h3>Methods</h3><p>In our study, we conducted maxillary molar extractions in aged C57BL/6J and <em>App</em><sup><em>NL-G-F</em></sup> mice and observed their condition over a 3-month period. During this time, we documented significant behavioral and genetic differences between mice in the control groups and mice that underwent tooth extraction. Notably, mice that underwent tooth extraction exhibited a considerable decline in cognitive function and increased in aggression 3 months after tooth extraction compared with the control groups (C57BL/6J and <em>App</em><sup><em>NL-G-F</em></sup>mice).</p></div><div><h3>Results</h3><p>Our findings suggest that molar loss may lead to reduced 5-HT levels in the hippocampus, possibly mediated by the trigeminal nerve, contributing to the development of aggression and BPSD in AD.</p></div><div><h3>Conclusion</h3><p>This study sheds light on the intricate relationships between oral health, 5-HT, and AD symptoms, offering valuable insights into potential therapeutic avenues for managing BPSD in patients with dementia.</p></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1349007924000744/pdfft?md5=5ddb384ef8a89c918dfd65de96394830&pid=1-s2.0-S1349007924000744-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140194811","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}
Factors that induce bone formation during orthodontic tooth movement (OTM) remain unclear. Gli1 was recently identified as a stem cell marker in the periodontal ligament (PDL). Therefore, we evaluated the mechanism of differentiation of Cre/LoxP-mediated Gli1/Tomato+ cells into osteoblasts during OTM.
Methods
After the final administration of tamoxifen to 8-week-old Gli1-CreERT2/ROSA26-loxP-stop-loxP-tdTomato mice for 2 days, nickel–titanium closed coil springs were attached between the upper anterior alveolar bone and the first molar. Immunohistochemical localizations of β-catenin, Smad4, and Runx2 were observed in the PDL on 2, 5, and 10 days after OTM initiation.
Results
In the untreated tooth, few Gli1/Tomato+ cells were detected in the PDL. Two days after OTM initiation, the number of Gli1/Tomato+ cells increased in the PDL on the tension side. On this side, 49.3 ± 7.0% of β-catenin+ and 48.7 ± 5.7% of Smad4+ cells were found in the PDL, and Runx2 expression was detected in some Gli1/Tomato+ cells apart from the alveolar bone. The number of positive cells in the PDL reached a maximum on day 5. In contrast, on the compression side, β-catenin and Smad4 exhibited less immunoreactivity. On day 10, Gli1/Tomato+ cells were aligned on the alveolar bone on the tension side, with some expressing Runx2.
Conclusions
Gli1+ cells in the PDL differentiated into osteoblasts during OTM. Wnt and bone morphogenetic proteins signaling pathways may be involved in this differentiation.
{"title":"Osteoblast differentiation of Gli1⁺ cells via Wnt and BMP signaling pathways during orthodontic tooth movement","authors":"Yuri Seki , Hiroaki Takebe , Yuya Nakao , Kohei Sato , Toshihide Mizoguchi , Hiroaki Nakamura , Masahiro Iijima , Akihiro Hosoya","doi":"10.1016/j.job.2024.03.004","DOIUrl":"10.1016/j.job.2024.03.004","url":null,"abstract":"<div><h3>Objectives</h3><p>Factors that induce bone formation during orthodontic tooth movement (OTM) remain unclear. Gli1 was recently identified as a stem cell marker in the periodontal ligament (PDL). Therefore, we evaluated the mechanism of differentiation of Cre/LoxP-mediated Gli1/Tomato<sup>+</sup> cells into osteoblasts during OTM.</p></div><div><h3>Methods</h3><p>After the final administration of tamoxifen to 8-week-old Gli1-Cre<sup>ERT2</sup>/ROSA26-loxP-stop-loxP-tdTomato mice for 2 days, nickel–titanium closed coil springs were attached between the upper anterior alveolar bone and the first molar. Immunohistochemical localizations of β-catenin, Smad4, and Runx2 were observed in the PDL on 2, 5, and 10 days after OTM initiation.</p></div><div><h3>Results</h3><p>In the untreated tooth, few Gli1/Tomato<sup>+</sup> cells were detected in the PDL. Two days after OTM initiation, the number of Gli1/Tomato<sup>+</sup> cells increased in the PDL on the tension side. On this side, 49.3 ± 7.0% of β-catenin<sup>+</sup> and 48.7 ± 5.7% of Smad4<sup>+</sup> cells were found in the PDL, and Runx2 expression was detected in some Gli1/Tomato<sup>+</sup> cells apart from the alveolar bone. The number of positive cells in the PDL reached a maximum on day 5. In contrast, on the compression side, β-catenin and Smad4 exhibited less immunoreactivity. On day 10, Gli1/Tomato<sup>+</sup> cells were aligned on the alveolar bone on the tension side, with some expressing Runx2.</p></div><div><h3>Conclusions</h3><p>Gli1<sup>+</sup> cells in the PDL differentiated into osteoblasts during OTM. Wnt and bone morphogenetic proteins signaling pathways may be involved in this differentiation.</p></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140159190","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Rheumatoid arthritis (RA) is an autoimmune disease characterized by progressive joint destruction. Early diagnosis and treatment, before joint deformation or destruction occurs, are crucial. Identifying novel biomarkers for RA in saliva could potentially enable early detection of the disease, prior to its onset.
Methods
We conducted a comprehensive proteomic analysis of salivary proteins in a mouse model of RA. Proteins were identified using western blotting and enzyme-linked immunosorbent assay in the serum, saliva, and ankle joints of DBA/1JJmsSlc mice, a model of RA. Ankle joints and submandibular glands were stained with hematoxylin and eosin and immunostained, and the results were compared with those of control mice.
Results
Citrullinated alpha-1 antitrypsin (A1AT, 46 kDa) was commonly detected in the saliva, serum, and ankle joints of mice with severe RA and was confirmed through proteomic analysis. Western blotting showed a band corresponding to 46 kDa in the serum, saliva, and ankle joints. Immunostaining of the ankle joints with the A1AT antibody showed a strong positive signal in the synovium.
Conclusions
In DBA/1JJmsSlc mice, cyclic citrullinated peptide antibodies and A1AT may be involved in citrullination and contribute to the development and severity of RA, making them valuable treatment targets requiring further study.
目的：类风湿性关节炎（RA）是一种以进行性关节破坏为特征的自身免疫性疾病。在关节变形或破坏发生之前进行早期诊断和治疗至关重要。鉴定唾液中的新型类风湿性关节炎生物标志物有可能在疾病发作前对其进行早期检测：我们对RA小鼠模型的唾液蛋白质进行了全面的蛋白质组学分析。我们使用 Western 印迹法和酶联免疫吸附法鉴定了 DBA/1JJmsSlc 小鼠（一种 RA 模型）血清、唾液和踝关节中的蛋白质。对踝关节和颌下腺进行苏木精和伊红染色及免疫染色，并将结果与对照组小鼠进行比较：结果：在严重RA小鼠的唾液、血清和踝关节中普遍检测到瓜氨酸化α-1抗胰蛋白酶（A1AT，46 kDa），并通过蛋白质组分析得到证实。Western 印迹显示，在血清、唾液和踝关节中都有一条与 46 kDa 相对应的条带。用 A1AT 抗体对踝关节进行免疫染色显示，滑膜中出现了强烈的阳性信号：结论：在DBA/1JJmsSlc小鼠中，环瓜氨酸肽抗体和A1AT可能参与瓜氨酸化，并导致RA的发生和严重程度，因此是有价值的治疗靶点，需要进一步研究。
{"title":"Identification of citrullinated α1-antitrypsin (A1AT) in saliva in a mouse model of rheumatoid arthritis","authors":"Wakako Sakaguchi , Juri Saruta , Yuko Yamamoto , Tomoko Shimizu , Shinya Fuchida , Keiichi Tsukinoki","doi":"10.1016/j.job.2024.03.007","DOIUrl":"10.1016/j.job.2024.03.007","url":null,"abstract":"<div><h3>Objectives</h3><p>Rheumatoid arthritis (RA) is an autoimmune disease characterized by progressive joint destruction. Early diagnosis and treatment, before joint deformation or destruction occurs, are crucial. Identifying novel biomarkers for RA in saliva could potentially enable early detection of the disease, prior to its onset.</p></div><div><h3>Methods</h3><p>We conducted a comprehensive proteomic analysis of salivary proteins in a mouse model of RA. Proteins were identified using western blotting and enzyme-linked immunosorbent assay in the serum, saliva, and ankle joints of DBA/1JJmsSlc mice, a model of RA. Ankle joints and submandibular glands were stained with hematoxylin and eosin and immunostained, and the results were compared with those of control mice.</p></div><div><h3>Results</h3><p>Citrullinated alpha-1 antitrypsin (A1AT, 46 kDa) was commonly detected in the saliva, serum, and ankle joints of mice with severe RA and was confirmed through proteomic analysis. Western blotting showed a band corresponding to 46 kDa in the serum, saliva, and ankle joints. Immunostaining of the ankle joints with the A1AT antibody showed a strong positive signal in the synovium.</p></div><div><h3>Conclusions</h3><p>In DBA/1JJmsSlc mice, cyclic citrullinated peptide antibodies and A1AT may be involved in citrullination and contribute to the development and severity of RA, making them valuable treatment targets requiring further study.</p></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140330215","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The development of bio-three-dimensional (bio-3D) printers has led to significant advances in regenerative medicine. Three-dimensional constructs, including spheroids, are maintained by extracellular matrix proteins secreted by cells so that the cells can be cultured in conditions closer to the physiological environment. This study aimed to create a useful 3D construct as a model of the dentin-pulp complex.
Methods
We examined the expression patterns of extracellular matrix proteins and cell proliferation areas in a 3D construct created using O9-1 cells derived from cranial neural crest cells of mice. The 3D construct was created by sticking the spheroid cultures onto a needle array using a bio-3D printer.
Results
Cell proliferation areas along with characteristic expression of tenascin C and DMP1 were evaluated. The expression of tenascin C and DMP1 was significantly enhanced in the spheroids compared to that in two-dimensional cultures. Moreover, cell proliferation regions and tenascin C expression were confirmed in the outer layer of spheroids in the embryonic stem cell medium, with insignificant DMP1 expression being observed. Interestingly, in a 3D construct cultured in calcification-induction medium, DMP1 expression was promoted, and DMP1-positive cells existed in the outermost layer without overlapping with tenascin C expression.
Conclusions
The extracellular matrix proteins, tenascin C and DMP1, were expressed in a polarized manner in spheroids and 3D constructs, similar to the findings in the dental papilla. Therefore, these 3D constructs show potential as artificial models for studying odontogenesis.
目的：生物三维（bio-3D）打印机的开发为再生医学带来了重大进展。三维构建体（包括球体）由细胞分泌的细胞外基质蛋白维持，这样细胞就能在更接近生理环境的条件下进行培养。本研究旨在创建一种有用的三维构建体，作为牙本质-牙髓复合体的模型 方法：我们研究了使用小鼠颅神经嵴细胞衍生的 O9-1 细胞创建的三维构建体中细胞外基质蛋白的表达模式和细胞增殖区域。三维构建体是用生物三维打印机将球形培养物粘贴在针阵列上制作而成的：结果：对细胞增殖区域以及tenascin C和DMP1的特征表达进行了评估。与二维培养物相比，球形培养物中强筋蛋白 C 和 DMP1 的表达明显增强。此外，在胚胎干细胞培养基中，细胞增殖区和腱鞘蛋白 C 的表达在球形体外层得到了证实，而 DMP1 的表达却不明显。有趣的是，在钙化诱导培养基中培养的三维构建体中，DMP1的表达得到了促进，DMP1阳性细胞存在于最外层，与tenascin C的表达没有重叠：结论：细胞外基质蛋白tenascin C和DMP1在球体和三维构建体中以极化方式表达，这与在牙乳头中的发现相似。因此，这些三维构建体显示出作为研究牙体发生的人工模型的潜力。
{"title":"Creating 3D constructs with cranial neural crest-derived cell lines using a bio-3D printer","authors":"Masahide Taguchi , Shohei Yoshimoto , Kanako Suyama , Satoko Sumi , Shirabe Ohki , Kayoko Ogata , Ryota Fujimoto , Daiki Murata , Koichi Nakayama , Kyoko Oka","doi":"10.1016/j.job.2024.05.005","DOIUrl":"10.1016/j.job.2024.05.005","url":null,"abstract":"<div><h3>Objectives</h3><p>The development of bio-three-dimensional (bio-3D) printers has led to significant advances in regenerative medicine. Three-dimensional constructs, including spheroids, are maintained by extracellular matrix proteins secreted by cells so that the cells can be cultured in conditions closer to the physiological environment. This study aimed to create a useful 3D construct as a model of the dentin-pulp complex.</p></div><div><h3>Methods</h3><p>We examined the expression patterns of extracellular matrix proteins and cell proliferation areas in a 3D construct created using O9-1 cells derived from cranial neural crest cells of mice. The 3D construct was created by sticking the spheroid cultures onto a needle array using a bio-3D printer.</p></div><div><h3>Results</h3><p>Cell proliferation areas along with characteristic expression of tenascin C and DMP1 were evaluated. The expression of tenascin C and DMP1 was significantly enhanced in the spheroids compared to that in two-dimensional cultures. Moreover, cell proliferation regions and tenascin C expression were confirmed in the outer layer of spheroids in the embryonic stem cell medium, with insignificant DMP1 expression being observed. Interestingly, in a 3D construct cultured in calcification-induction medium, DMP1 expression was promoted, and DMP1-positive cells existed in the outermost layer without overlapping with tenascin C expression.</p></div><div><h3>Conclusions</h3><p>The extracellular matrix proteins, tenascin C and DMP1, were expressed in a polarized manner in spheroids and 3D constructs, similar to the findings in the dental papilla. Therefore, these 3D constructs show potential as artificial models for studying odontogenesis.</p></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1349007924000884/pdfft?md5=8f885124196ccc1de6d2898988bc6099&pid=1-s2.0-S1349007924000884-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140945462","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}
This systematic review aimedto evaluate the remineralizing efficacy of calcium sucrose phosphate (CaSP) for the treatment of white spot lesions (WSLs) that commonly occur after orthodontic treatment with fixed appliances using various randomized controlled trials (RCTs) available in the literature todate.
Highlights
Adhering to the Preferred Reporting Items for Systematic Reviews and Meta-Analyzes (PRISMA) guidelines, RCTs that assessed the efficacious remineralizing potential of CaSP on WSLs and demineralized enamel and compared it with either no intervention or other remineralizing agents wereselected. The methodological rigor of the included studies was subjected to the Risk of Bias tool-2 (ROB-2) and Grading of Recommendations, Assessment, Development and Evaluation (GRADE) tools. Furthermore, a Begg's Funnel Plot was used to assess publication bias. The qualitative analysis encompassed a corpus of 36 studies. The remineralization potential of CaSP was investigated using an array of parameters, including surface microhardness, surface morphology, surface roughness, mineral content, and lesion size and depth. Based on the ROB-2 tool, most of the included studies were judged to be high risk, largely attributable to the presence of attrition bias. Using the GRADE framework, the certainty of evidence was determined to be moderate.
Conclusion
This systematic review reveals that CaSP yields favorableoutcomes in terms of increased surface microhardness and calcium-phosphate content, reduced demineralized area and surface roughness, and enhanced surface topography
{"title":"Remineralizing potential of Calcium Sucrose Phosphate in white spot lesions: A Systematic Review","authors":"Tanisha Rout, Amol Patil, Sonakashee Deshmukh, Sonakshi Sharma","doi":"10.1016/j.job.2024.04.005","DOIUrl":"10.1016/j.job.2024.04.005","url":null,"abstract":"<div><h3>Background</h3><p>This systematic review aimedto evaluate the remineralizing efficacy of calcium sucrose phosphate (CaSP) for the treatment of white spot lesions (WSLs) that commonly occur after orthodontic treatment with fixed appliances using various randomized controlled trials (RCTs) available in the literature todate.</p></div><div><h3>Highlights</h3><p>Adhering to the Preferred Reporting Items for Systematic Reviews and Meta-Analyzes (PRISMA) guidelines, RCTs that assessed the efficacious remineralizing potential of CaSP on WSLs and demineralized enamel and compared it with either no intervention or other remineralizing agents wereselected. The methodological rigor of the included studies was subjected to the Risk of Bias tool-2 (ROB-2) and Grading of Recommendations, Assessment, Development and Evaluation (GRADE) tools. Furthermore, a Begg's Funnel Plot was used to assess publication bias. The qualitative analysis encompassed a corpus of 36 studies. The remineralization potential of CaSP was investigated using an array of parameters, including surface microhardness, surface morphology, surface roughness, mineral content, and lesion size and depth. Based on the ROB-2 tool, most of the included studies were judged to be high risk, largely attributable to the presence of attrition bias. Using the GRADE framework, the certainty of evidence was determined to be moderate.</p></div><div><h3>Conclusion</h3><p>This systematic review reveals that CaSP yields favorableoutcomes in terms of increased surface microhardness and calcium-phosphate content, reduced demineralized area and surface roughness, and enhanced surface topography</p></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140762573","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-06-01DOI: 10.1016/j.job.2024.04.007
Juan Du , Wei Zhou , Zhe Sun , Weilong Zhang , Wei Luo , Shanshan Liu
Objectives
Osteoporosis is the most common metabolic bone disease worldwide. The decrease in bone mass is primarily accompanied by a decrease in the number and activity of osteoblasts. Peroxiredoxins (PRDXs) are proteins that detect extremely low peroxide levels and act as sensors that regulate oxidation signals, thereby regulating various cellular functions. This study aimed to evaluate the effects of PRDX1 and estrogen on the biological behavior of osteoblasts, including their proliferation and differentiation.
Methods
Ovariectomized (OVX) mice were used to establish a model of osteoporosis and perform morphological and immunohistochemical analyses. Prdx1 gene knockout and overexpression were performed in mouse MC3T3-E1 pre-osteoblasts to assess proliferation and osteogenic differentiation using the cell counting kit-8, quantitative reverse transcription polymerase chain reaction, western blotting (WB), Alizarin Red S staining, etc.
Results
The OVX mice exhibited osteoporosis and PRDX1 expression increased. In vitro experiments showed that during the osteogenic differentiation of osteoblasts, PRDX1 expression decreased, while the expression of COL1 and RUNX2 increased. After Prdx1 knockout, the proliferation of osteoblasts decreased; expression of Runx2, ALP, and COL1 increased; and mineralization increased. However, after Prdx1 overexpression, osteoblast proliferation was enhanced, whereas osteogenic differentiation and mineralization were inhibited. Estrogen inhibits the H2O2-induced decrease in osteoblastic differentiation and increase in PRDX1 expression. WB revealed that when LY294002 inhibited the AKT signaling pathway, the levels of p-AKT1, p-P65, and PRDX1 protein in MC3T3-E1 cells decreased. However, when pyrrolidine dithiocarbamate (PDTC) inhibited the NF-κB signaling pathway, the expression of p-AKT1 and PRDX1 did not change except for a significant reduction of p-P65 expression. Furthermore, PDTC reversed the decreased expression of RUNX2, ALP, and COL1 caused by PRDX1 overexpression.
Conclusions
PRDX1 promotes the proliferation of osteoblasts and inhibits osteogenic differentiation. Estrogen regulated osteoblastic differentiation by affecting the expression of PRDX1 in osteoblasts, and the effect is related to the AKT1/NF-κB signaling pathway.
{"title":"Peroxiredoxin 1 promotes proliferation and inhibits differentiation of MC3T3-E1 cells via AKT1 / NF-κB signaling pathway","authors":"Juan Du , Wei Zhou , Zhe Sun , Weilong Zhang , Wei Luo , Shanshan Liu","doi":"10.1016/j.job.2024.04.007","DOIUrl":"10.1016/j.job.2024.04.007","url":null,"abstract":"<div><h3>Objectives</h3><p>Osteoporosis is the most common metabolic bone disease worldwide. The decrease in bone mass is primarily accompanied by a decrease in the number and activity of osteoblasts. Peroxiredoxins (PRDXs) are proteins that detect extremely low peroxide levels and act as sensors that regulate oxidation signals, thereby regulating various cellular functions. This study aimed to evaluate the effects of PRDX1 and estrogen on the biological behavior of osteoblasts, including their proliferation and differentiation.</p></div><div><h3>Methods</h3><p>Ovariectomized (OVX) mice were used to establish a model of osteoporosis and perform morphological and immunohistochemical analyses. <em>Prdx1</em> gene knockout and overexpression were performed in mouse MC3T3-E1 pre-osteoblasts to assess proliferation and osteogenic differentiation using the cell counting kit-8, quantitative reverse transcription polymerase chain reaction, western blotting (WB), Alizarin Red S staining, etc.</p></div><div><h3>Results</h3><p>The OVX mice exhibited osteoporosis and PRDX1 expression increased. <em>In vitro</em> experiments showed that during the osteogenic differentiation of osteoblasts, PRDX1 expression decreased, while the expression of COL1 and RUNX2 increased. After <em>Prdx1</em> knockout, the proliferation of osteoblasts decreased; expression of Runx2, ALP, and COL1 increased; and mineralization increased. However, after <em>Prdx1</em> overexpression, osteoblast proliferation was enhanced, whereas osteogenic differentiation and mineralization were inhibited. Estrogen inhibits the H<sub>2</sub>O<sub>2</sub>-induced decrease in osteoblastic differentiation and increase in PRDX1 expression. WB revealed that when LY294002 inhibited the AKT signaling pathway, the levels of <em>p</em>-AKT1, p-P65, and PRDX1 protein in MC3T3-E1 cells decreased. However, when pyrrolidine dithiocarbamate (PDTC) inhibited the NF-κB signaling pathway, the expression of <em>p</em>-AKT1 and PRDX1 did not change except for a significant reduction of p-P65 expression. Furthermore, PDTC reversed the decreased expression of RUNX2, ALP, and COL1 caused by PRDX1 overexpression.</p></div><div><h3>Conclusions</h3><p>PRDX1 promotes the proliferation of osteoblasts and inhibits osteogenic differentiation. Estrogen regulated osteoblastic differentiation by affecting the expression of PRDX1 in osteoblasts, and the effect is related to the AKT1/NF-κB signaling pathway.</p></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140768176","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Typical agonists of G protein-coupled receptors (GPCRs), including muscarinic acetylcholine receptors (mAChRs), activate both G-protein and β-arrestin signaling systems, and are termed balanced agonists. In contrast, biased agonists selectively activate a single pathway, thereby offering therapeutic potential for the specific activation of that pathway. The mAChR agonists carbachol and pilocarpine are known to induce phosphorylation of extracellular signal-regulated kinase-1/2 (ERK1/2) via G-protein-dependent and -independent pathways, respectively. We investigated the involvement of β-arrestin and its downstream mechanisms in the ERK1/2 phosphorylation induced by carbachol and pilocarpine in the human salivary ductal cell line, HSY cells.
Methods
HSY cells were stimulated with pilocarpine or carbachol, with or without various inhibitors. The cell lysates were analyzed by western blotting using the antibodies p44/p42MAPK and phosphor-p44/p42MAPK.
Results
Western blot analysis revealed that carbachol elicited greater stimulation of ERK1/2 phosphorylation compared to pilocarpine. ERK1/2 phosphorylation was inhibited by atropine and gefitinib, suggesting that mAChR activation induces transactivation of epidermal growth factor receptors (EGFR). Moreover, inhibition of carbachol-mediated ERK1/2 phosphorylation was achieved by GF-109203X (a PKC inhibitor), a βARK1/GRK2 inhibitor, barbadin (a β-arrestin inhibitor), pitstop 2 (a clathrin inhibitor), and dynole 34-2 (a dynamin inhibitor). In contrast, pilocarpine-mediated ERK1/2 phosphorylation was only inhibited by barbadin (a β-arrestin inhibitor) and PP2 (a Src inhibitor).
Conclusion
Carbachol activates both G-protein and β-arrestin pathways, whereas pilocarpine exclusively activates the β-arrestin pathway. Additionally, downstream of β-arrestin, carbachol activates clathrin-dependent internalization, while pilocarpine activates Src.
目的：G 蛋白偶联受体（GPCRs）（包括毒蕈碱乙酰胆碱受体（mAChRs））的典型激动剂可同时激活 G 蛋白和 β-restin 信号系统，因此被称为平衡激动剂。相比之下，偏性激动剂选择性地激活单一途径，从而为特定途径的激活提供治疗潜力。众所周知，mAChR 激动剂卡巴胆碱（carbachol）和皮洛卡品（pilocarpine）可分别通过依赖 G 蛋白和不依赖 G 蛋白的途径诱导细胞外信号调节激酶-1/2（ERK1/2）磷酸化。我们研究了β-arrestin及其下游机制在卡巴胆碱和皮洛卡品诱导人唾液腺导管细胞系HSY细胞ERK1/2磷酸化中的参与情况。用 p44/p42MAPK 和 phosphor-p44/p42MAPK 抗体对细胞裂解液进行 Western 印迹分析：Western 印迹分析显示，与皮洛卡品相比，卡巴胆碱对 ERK1/2 磷酸化的刺激更大。阿托品和吉非替尼抑制了ERK1/2的磷酸化，这表明mAChR的激活诱导了表皮生长因子受体（EGFR）的转激活。此外，GF-109203X（PKC 抑制剂）、βARK1/GRK2 抑制剂、barbadin（β-arrestin 抑制剂）、pitstop 2（clathrin 抑制剂）和 dynole 34-2（dynamin 抑制剂）都能抑制卡巴胆碱介导的 ERK1/2 磷酸化。相反，皮洛卡品介导的ERK1/2磷酸化只受到barbadin（一种β-阿司匹林抑制剂）和PP2（一种Src抑制剂）的抑制：结论：卡巴胆碱同时激活G蛋白和β-阿司匹林通路，而皮洛卡品只激活β-阿司匹林通路。此外，在 β-阿司匹林的下游，卡巴胆碱激活依附于凝集素的内化，而皮洛卡品激活 Src。
{"title":"Muscarinic acetylcholine receptor-mediated phosphorylation of extracellular signal-regulated kinase in HSY salivary ductal cells involves distinct signaling pathways","authors":"Rezon Yanuar, Shingo Semba, Akihiro Nezu, Akihiko Tanimura","doi":"10.1016/j.job.2024.02.002","DOIUrl":"10.1016/j.job.2024.02.002","url":null,"abstract":"<div><h3>Objectives</h3><p>Typical agonists of G protein-coupled receptors (GPCRs), including muscarinic acetylcholine receptors (mAChRs), activate both G-protein and β-arrestin signaling systems, and are termed balanced agonists. In contrast, biased agonists selectively activate a single pathway, thereby offering therapeutic potential for the specific activation of that pathway. The mAChR agonists carbachol and pilocarpine are known to induce phosphorylation of extracellular signal-regulated kinase-1/2 (ERK1/2) <em>via</em> G-protein-dependent and -independent pathways, respectively. We investigated the involvement of β-arrestin and its downstream mechanisms in the ERK1/2 phosphorylation induced by carbachol and pilocarpine in the human salivary ductal cell line, HSY cells.</p></div><div><h3>Methods</h3><p>HSY cells were stimulated with pilocarpine or carbachol, with or without various inhibitors. The cell lysates were analyzed by western blotting using the antibodies p44/p42<sup>MAPK</sup> and phosphor-p44/p42<sup>MAPK</sup>.</p></div><div><h3>Results</h3><p>Western blot analysis revealed that carbachol elicited greater stimulation of ERK1/2 phosphorylation compared to pilocarpine. ERK1/2 phosphorylation was inhibited by atropine and gefitinib, suggesting that mAChR activation induces transactivation of epidermal growth factor receptors (EGFR). Moreover, inhibition of carbachol-mediated ERK1/2 phosphorylation was achieved by GF-109203X (a PKC inhibitor), a βARK1/GRK2 inhibitor, barbadin (a β-arrestin inhibitor), pitstop 2 (a clathrin inhibitor), and dynole 34-2 (a dynamin inhibitor). In contrast, pilocarpine-mediated ERK1/2 phosphorylation was only inhibited by barbadin (a β-arrestin inhibitor) and PP2 (a Src inhibitor).</p></div><div><h3>Conclusion</h3><p>Carbachol activates both G-protein and β-arrestin pathways, whereas pilocarpine exclusively activates the β-arrestin pathway. Additionally, downstream of β-arrestin, carbachol activates clathrin-dependent internalization, while pilocarpine activates Src.</p></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139713228","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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