Pub Date : 2025-01-15DOI: 10.1016/j.job.2025.100614
Su-Kyung Son, Jung-Sun Moon, Dong-Wook Yang, Na-Ri Jung, Jee-Hae Kang, Bin-Na Lee, Sun-Hun Kim, Min-Seok Kim
Objectives
We investigated the involvement of FOXO3a in lipopolysaccharide (LPS)-induced inflammation in primary human dental pulp cells (HDPCs).
Methods
HDPCs that were isolated from donors undergoing tooth extraction for orthodontic purposes were cultured with or without 1 μg/mL LPS at various intervals. The FOXO3a localization in the HDPCs was verified using immunofluorescence. Proinflammatory cytokines, such as interleukin (IL) 1β, IL6, and IL8, as well as their underlying mechanisms were assessed by observing gene and protein expressions through quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analyses.
Results
LPS treatment enhanced the expressions of IL1β, IL6, and IL8 in HDPCs, concurrently activating nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB). Furthermore, FOXO3a expression was higher in the LPS-stimulated HDPCs, as confirmed by immunofluorescence localization. The results of loss-/gain-of-function approaches confirmed the regulatory role of FOXO3a in inflammatory HDPCs. FOXO3a knockdown attenuated proinflammatory cytokine expression; FOXO3a overexpression augmented their expression levels. FOXO3a inhibited retinoic acid receptor-related orphan receptor alpha (RORα) expression, thereby inactivating NFκB.
Conclusion
Our findings suggest that FOXO3a contributes to homeostasis in HDPCs through modulating the expression of proinflammatory cytokines under inflammatory conditions.
{"title":"Role of FOXO3a in LPS-induced inflammatory conditions in human dental pulp cells","authors":"Su-Kyung Son, Jung-Sun Moon, Dong-Wook Yang, Na-Ri Jung, Jee-Hae Kang, Bin-Na Lee, Sun-Hun Kim, Min-Seok Kim","doi":"10.1016/j.job.2025.100614","DOIUrl":"10.1016/j.job.2025.100614","url":null,"abstract":"<div><h3>Objectives</h3><div>We investigated the involvement of FOXO3a in lipopolysaccharide (LPS)-induced inflammation in primary human dental pulp cells (HDPCs).</div></div><div><h3>Methods</h3><div>HDPCs that were isolated from donors undergoing tooth extraction for orthodontic purposes were cultured with or without 1 μg/mL LPS at various intervals. The FOXO3a localization in the HDPCs was verified using immunofluorescence. Proinflammatory cytokines, such as interleukin (IL) 1β, IL6, and IL8, as well as their underlying mechanisms were assessed by observing gene and protein expressions through quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analyses.</div></div><div><h3>Results</h3><div>LPS treatment enhanced the expressions of IL1β, IL6, and IL8 in HDPCs, concurrently activating nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB). Furthermore, FOXO3a expression was higher in the LPS-stimulated HDPCs, as confirmed by immunofluorescence localization. The results of loss-/gain-of-function approaches confirmed the regulatory role of FOXO3a in inflammatory HDPCs. FOXO3a knockdown attenuated proinflammatory cytokine expression; FOXO3a overexpression augmented their expression levels. FOXO3a inhibited retinoic acid receptor-related orphan receptor alpha (RORα) expression, thereby inactivating NFκB.</div></div><div><h3>Conclusion</h3><div>Our findings suggest that FOXO3a contributes to homeostasis in HDPCs through modulating the expression of proinflammatory cytokines under inflammatory conditions.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 1","pages":"Article 100614"},"PeriodicalIF":2.6,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143013840","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 : 2025-01-11DOI: 10.1016/j.job.2025.100612
Mako Sakakibara , Tomoka Hasegawa , Mai Haraguchi-Kitakamae , Yan Shi , Weisong Li , Jiaxin Cui , Xuanyu Liu , Tomomaya Yamamoto , Hiromi Hongo , Norio Amizuka , Yoshiaki Sato , Masanori Kikuchi
Objectives
Hydroxyapatite (HAp)/collagen (Col) cylinders with laminated collagen layers were implanted into the tibial diaphysis of rats and examined histochemically to clarify how the orientation of HAp and Col bone-like nanocomposite fibers in HAp/Col blocks affects bone resorption and formation.
Methods
HAp/Col fibers were synthesized and compressed into cylindrical blocks to mimic bone nanostructures. These were implanted into the cortical bone cavities of 10-week-old male Wistar rats with fiber bundles parallel to the tibial surface. The implants were histologically analyzed at 3, 5, 7, 14, and 28 days after implantation.
Results
TRAP-positive osteoclasts appeared after 3–5 days in the lateral region of the graft, where the fiber ends were exposed, but not in the bottom region, where the HAp/Col fibers were parallel to the surface. Osteoclasts were observed in both regions by day 14. PHOSPHO1-positive osteoblasts were first detected on day 5, appearing slightly away from the cylinder laterally but directly on the bottom surface. A few osteoblasts contacted the block laterally, whereas many were observed on the new bone tissue at the bottom, between days 7 and 14. Bone formation was induced earlier in the bottom region, whereas lateral resorption was dominant. This suggested the uncoupling of bone resorption and formation in the early postimplantation stages. However, bone remodeling shifted to coupling between osteoclasts and osteoblasts throughout the cylinder by day 28.
Conclusion
The orientation of HAp/Col fibers in HAp/Col graft materials substantially affected the preferential induction of bone resorption or formation during the early stages of bone regeneration.
{"title":"Histochemical analysis of osteoclast and osteoblast distributions on hydroxyapatite/collagen bone-like nanocomposite embedded in rat tibiae","authors":"Mako Sakakibara , Tomoka Hasegawa , Mai Haraguchi-Kitakamae , Yan Shi , Weisong Li , Jiaxin Cui , Xuanyu Liu , Tomomaya Yamamoto , Hiromi Hongo , Norio Amizuka , Yoshiaki Sato , Masanori Kikuchi","doi":"10.1016/j.job.2025.100612","DOIUrl":"10.1016/j.job.2025.100612","url":null,"abstract":"<div><h3>Objectives</h3><div>Hydroxyapatite (HAp)/collagen (Col) cylinders with laminated collagen layers were implanted into the tibial diaphysis of rats and examined histochemically to clarify how the orientation of HAp and Col bone-like nanocomposite fibers in HAp/Col blocks affects bone resorption and formation.</div></div><div><h3>Methods</h3><div>HAp/Col fibers were synthesized and compressed into cylindrical blocks to mimic bone nanostructures. These were implanted into the cortical bone cavities of 10-week-old male Wistar rats with fiber bundles parallel to the tibial surface. The implants were histologically analyzed at 3, 5, 7, 14, and 28 days after implantation.</div></div><div><h3>Results</h3><div>TRAP-positive osteoclasts appeared after 3–5 days in the lateral region of the graft, where the fiber ends were exposed, but not in the bottom region, where the HAp/Col fibers were parallel to the surface. Osteoclasts were observed in both regions by day 14. PHOSPHO1-positive osteoblasts were first detected on day 5, appearing slightly away from the cylinder laterally but directly on the bottom surface. A few osteoblasts contacted the block laterally, whereas many were observed on the new bone tissue at the bottom, between days 7 and 14. Bone formation was induced earlier in the bottom region, whereas lateral resorption was dominant. This suggested the uncoupling of bone resorption and formation in the early postimplantation stages. However, bone remodeling shifted to coupling between osteoclasts and osteoblasts throughout the cylinder by day 28.</div></div><div><h3>Conclusion</h3><div>The orientation of HAp/Col fibers in HAp/Col graft materials substantially affected the preferential induction of bone resorption or formation during the early stages of bone regeneration.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 1","pages":"Article 100612"},"PeriodicalIF":2.6,"publicationDate":"2025-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142980205","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}
Temporomandibular joint (TMJ) osteoarthritis (OA) is an inflammatory disease that involves periarthritis of the TMJ and destruction of cartilage tissue in the mandibular condyle. However, the role of proinflammatory cytokines in the expression levels of matrix metalloproteinase (MMP) remains inconclusive. Thus, in this study, we aimed to investigate the effect of proinflammatory cytokines on the expression of MMPs.
Methods
FLS1 cells (mouse TMJ-derived synovial cell line) were treated with tumor necrosis factor alpha (TNF-α) or interleukin (IL)-1β in the presence or absence of mitogen-activated protein kinase (MAPK) inhibitors. The mRNA expression levels of MMP-2 and MMP-9 were examined by reverse transcription-quantitative polymerase chain reaction. Additionally, the phosphorylation status of extracellular signal-regulated kinase (ERK)1/2 and p38 MAPK in the FLS1 cells treated with TNF-α or IL-1β was evaluated by performing western blotting analysis.
Results
TNF-α and IL-1β significantly increased the expression of MMP-9 in the FLS1 cells; however, MMP-2 expression remained unaffected. Mitogen-activated protein kinase kinase (MEK) and p38 MAPK inhibitors significantly suppressed cytokine-induced MMP-9 upregulation. Conversely, Jun amino-terminal kinase (JNK) inhibitors further increased MMP-9 expression in the cells treated with TNF-α or IL-1β. Moreover, TNF-α and IL-1β enhanced ERK1/2 and p38 MAPK phosphorylation in the FLS1 cells.
Conclusions
TNF-α and IL-1β induced MMP-9 expression in the FLS1 cells via the MEK/ERK and p38 MAPK pathways and suppressed it via the JNK pathway. Thus, proinflammatory cytokines control MMP-9 expression in TMJ-OA by regulating multiple MAPK pathways.
{"title":"Proinflammatory cytokine-induced matrix metalloproteinase-9 expression in temporomandibular joint osteoarthritis is regulated by multiple intracellular mitogen-activated protein kinase pathways","authors":"Karen Abe , Seiji Yokota , Shikino Matsumoto , Hayato Ujiie , Emiko Kikuchi , Kazuro Satoh , Akira Ishisaki , Naoyuki Chosa","doi":"10.1016/j.job.2024.100609","DOIUrl":"10.1016/j.job.2024.100609","url":null,"abstract":"<div><h3>Objectives</h3><div>Temporomandibular joint (TMJ) osteoarthritis (OA) is an inflammatory disease that involves periarthritis of the TMJ and destruction of cartilage tissue in the mandibular condyle. However, the role of proinflammatory cytokines in the expression levels of matrix metalloproteinase (MMP) remains inconclusive. Thus, in this study, we aimed to investigate the effect of proinflammatory cytokines on the expression of MMPs.</div></div><div><h3>Methods</h3><div>FLS1 cells (mouse TMJ-derived synovial cell line) were treated with tumor necrosis factor alpha (TNF-α) or interleukin (IL)-1β in the presence or absence of mitogen-activated protein kinase (MAPK) inhibitors. The mRNA expression levels of MMP-2 and MMP-9 were examined by reverse transcription-quantitative polymerase chain reaction. Additionally, the phosphorylation status of extracellular signal-regulated kinase (ERK)1/2 and p38 MAPK in the FLS1 cells treated with TNF-α or IL-1β was evaluated by performing western blotting analysis.</div></div><div><h3>Results</h3><div>TNF-α and IL-1β significantly increased the expression of MMP-9 in the FLS1 cells; however, MMP-2 expression remained unaffected. Mitogen-activated protein kinase kinase (MEK) and p38 MAPK inhibitors significantly suppressed cytokine-induced MMP-9 upregulation. Conversely, Jun amino-terminal kinase (JNK) inhibitors further increased MMP-9 expression in the cells treated with TNF-α or IL-1β. Moreover, TNF-α and IL-1β enhanced ERK1/2 and p38 MAPK phosphorylation in the FLS1 cells.</div></div><div><h3>Conclusions</h3><div>TNF-α and IL-1β induced MMP-9 expression in the FLS1 cells via the MEK/ERK and p38 MAPK pathways and suppressed it via the JNK pathway. Thus, proinflammatory cytokines control MMP-9 expression in TMJ-OA by regulating multiple MAPK pathways.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 1","pages":"Article 100609"},"PeriodicalIF":2.6,"publicationDate":"2025-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142928232","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}
To investigate the effects of hypoxia on tooth germ development in mice and explore the underlying mechanisms.
Methods
Tooth germs were extracted from E14.5 mouse embryos and divided into the control and hypoxia groups for organ culture. The hypoxia group was exposed to hypoxia (0% oxygen) for 3 h, followed by normoxia for 21 h. After 2 or 7 days, samples were collected for morphometric analysis, reverse transcription-quantitative polymerase chain reaction, immunohistochemistry (IHC), and immunofluorescent staining (IF). Additionally, superoxide dismutase 3 (SOD3) expression patterns in mandibular molar tooth germs from C57BL/6 mouse embryos were analyzed using IHC. The SOD inhibitor sodium N, N-diethyldithiocarbamate trihydrate (DETC; 400 μM) was applied under normoxia for 3 days, followed by morphometry, IHC, and IF.
Results
After 7 days, the hypoxia group exhibited significantly smaller tooth size, fewer cusps, reduced cell proliferation, and increased apoptosis in the epithelium compared to the control group. Sod3 mRNA expression was higher than other Sod family member expressions in the control group. In the hypoxia group, Sod3 mRNA and SOD3 protein expression were significantly decreased, whereas hypoxia-inducible factor-1 expression and reactive oxygen species levels were increased. SOD3 was primarily expressed in the dental epithelium from E12.5 to E17.5. DETC impaired tooth germ development in the control group, resulting in a phenotype similar to that of the hypoxia group, and significantly reduced amelogenin and msh homeobox 2 expression in the epithelium.
Conclusions
Hypoxia impairs tooth germ development. SOD3 probably plays a protective role during this process.
{"title":"Possible role of superoxide dismutase 3 in hypoxia-induced developmental defects in murine molars","authors":"Yeming Lu, Yukiho Kobayashi, Yuki Niki, Keiji Moriyama","doi":"10.1016/j.job.2024.100611","DOIUrl":"10.1016/j.job.2024.100611","url":null,"abstract":"<div><h3>Objectives</h3><div>To investigate the effects of hypoxia on tooth germ development in mice and explore the underlying mechanisms.</div></div><div><h3>Methods</h3><div>Tooth germs were extracted from E14.5 mouse embryos and divided into the control and hypoxia groups for organ culture. The hypoxia group was exposed to hypoxia (0% oxygen) for 3 h, followed by normoxia for 21 h. After 2 or 7 days, samples were collected for morphometric analysis, reverse transcription-quantitative polymerase chain reaction, immunohistochemistry (IHC), and immunofluorescent staining (IF). Additionally, superoxide dismutase 3 (SOD3) expression patterns in mandibular molar tooth germs from C57BL/6 mouse embryos were analyzed using IHC. The SOD inhibitor sodium N, N-diethyldithiocarbamate trihydrate (DETC; 400 μM) was applied under normoxia for 3 days, followed by morphometry, IHC, and IF.</div></div><div><h3>Results</h3><div>After 7 days, the hypoxia group exhibited significantly smaller tooth size, fewer cusps, reduced cell proliferation, and increased apoptosis in the epithelium compared to the control group. <em>Sod3</em> mRNA expression was higher than other <em>Sod</em> family member expressions in the control group. In the hypoxia group, <em>Sod3</em> mRNA and SOD3 protein expression were significantly decreased, whereas hypoxia-inducible factor-1 expression and reactive oxygen species levels were increased. SOD3 was primarily expressed in the dental epithelium from E12.5 to E17.5. DETC impaired tooth germ development in the control group, resulting in a phenotype similar to that of the hypoxia group, and significantly reduced amelogenin and msh homeobox 2 expression in the epithelium.</div></div><div><h3>Conclusions</h3><div>Hypoxia impairs tooth germ development. SOD3 probably plays a protective role during this process.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 1","pages":"Article 100611"},"PeriodicalIF":2.6,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142923314","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 bacterium Porphyromonas gingivalis is a major causative agent of periodontitis. In this study, the anti-P. gingivalis compound in bilberry (Vaccinium myrtillus L.) was identified and its activity was compared with that of its related analogs.
Methods
An acetone-soluble bilberry fruit extract was purified using silica gel column chromatography, and the minimum inhibitory concentrations (MICs) of the purified fractions were determined against P. gingivalis. After purification, mass spectrometry, nuclear magnetic resonance, and optical rotation analyses were performed to identify the anti-P. gingivalis compounds. Furthermore, cell assays were performed to assess the anti-P. gingivalis activity and cytotoxicity of the identified compounds. The activity of these compounds was compared with that of their pentacyclic triterpene analogs.
Results
The anti-P. gingivalis in bilberry extracts was identified as ursolic acid, a pentacyclic triterpene (PCT). The MIC of ursolic acid against P. gingivalis was between 6.25 and 12.5 μg/mL; it killed P. gingivalis within 4 h of treatment at these concentrations. However, it showed no cytotoxicity against gingival carcinoma Ca9-22 cells at the MIC. Ursane-type PCT, including ursolic acid and oleanane-type PCT, exhibited anti-P. gingivalis activity.
Conclusions
Ursolic acid found in bilberry fruit extract exhibits anti-P. gingivalis activity. Similar activity is observed in a class of PCTs with a common structure.
{"title":"Characterization of the anti-Porphyromonas gingivalis compound in bilberry (Vaccinium myrtillus L.) and comparison with its analogs","authors":"Yutaroh Satoh , Kazuyuki Ishihara , Takaaki Kubota","doi":"10.1016/j.job.2024.100610","DOIUrl":"10.1016/j.job.2024.100610","url":null,"abstract":"<div><h3>Objectives</h3><div>The bacterium <em>Porphyromonas gingivalis</em> is a major causative agent of periodontitis. In this study, the anti-<em>P. gingivalis</em> compound in bilberry (<em>Vaccinium myrtillus</em> L.) was identified and its activity was compared with that of its related analogs.</div></div><div><h3>Methods</h3><div>An acetone-soluble bilberry fruit extract was purified using silica gel column chromatography, and the minimum inhibitory concentrations (MICs) of the purified fractions were determined against <em>P. gingivalis</em>. After purification, mass spectrometry, nuclear magnetic resonance, and optical rotation analyses were performed to identify the anti-<em>P. gingivalis</em> compounds. Furthermore, cell assays were performed to assess the anti-<em>P. gingivalis</em> activity and cytotoxicity of the identified compounds. The activity of these compounds was compared with that of their pentacyclic triterpene analogs.</div></div><div><h3>Results</h3><div>The anti-<em>P. gingivalis</em> in bilberry extracts was identified as ursolic acid, a pentacyclic triterpene (PCT). The MIC of ursolic acid against <em>P. gingivalis</em> was between 6.25 and 12.5 μg/mL; it killed <em>P. gingivalis</em> within 4 h of treatment at these concentrations. However, it showed no cytotoxicity against gingival carcinoma Ca9-22 cells at the MIC. Ursane-type PCT, including ursolic acid and oleanane-type PCT, exhibited anti-<em>P. gingivalis</em> activity.</div></div><div><h3>Conclusions</h3><div>Ursolic acid found in bilberry fruit extract exhibits anti-<em>P. gingivalis</em> activity. Similar activity is observed in a class of PCTs with a common structure.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 1","pages":"Article 100610"},"PeriodicalIF":2.6,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142923311","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 commensal microbiota of the finger-skin before and after ethanol disinfection were characterized and compared with the microbes isolated from the surface of smartphone touchscreens. The number of bacteria on the smartphone touchscreens was low, similar to that on the fingers after ethanol disinfection, suggesting that the surface of the touchscreens may not be suitable for the growth of microorganisms, rather than the surface of the fingers. Furthermore, ethanol disinfection reduced the number of bacteria on the finger-skin to 1/13 of the original count before disinfection.
{"title":"Profiling of the microbes on the surface of smartphone touchscreens","authors":"Nanase Takahashi , Anna Wakui , Yume Sekizawa , Miho Kawachi , Mirai Sekiguchi , Takashi Abe , Aya Sato , Misato Miyazawa , Manami Imai , Nagara Kaku , Shingo Maruyama , Hiroto Sano , Nahoko Kakihara , Jumpei Washio , Yuki Abiko , Kaori Tanaka , Nobuhiro Takahashi , Takuichi Sato","doi":"10.1016/j.job.2024.100607","DOIUrl":"10.1016/j.job.2024.100607","url":null,"abstract":"<div><div>The commensal microbiota of the finger-skin before and after ethanol disinfection were characterized and compared with the microbes isolated from the surface of smartphone touchscreens. The number of bacteria on the smartphone touchscreens was low, similar to that on the fingers after ethanol disinfection, suggesting that the surface of the touchscreens may not be suitable for the growth of microorganisms, rather than the surface of the fingers. Furthermore, ethanol disinfection reduced the number of bacteria on the finger-skin to 1/13 of the original count before disinfection.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 1","pages":"Article 100607"},"PeriodicalIF":2.6,"publicationDate":"2024-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142915892","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 cerebral cortex contains neurons that play a pivotal role in controlling rhythmic masticatory jaw movements. However, the population characteristics of individual cortical neuronal activity during mastication and the impact of tooth loss on these characteristics remain unclear. Thus, in this study, we aimed to determine the activity patterns of mastication-related motor cortical neurons elicited during mastication and examine the effects of tooth extraction on neuronal activity using two-photon Ca2+ imaging in head-restrained awake mice.
Methods
GCaMP6f-expressing adeno-associated virus serotype 1 was injected into the left motor cortex (centered 2 mm anterior and 2 mm lateral to the bregma) and electromyography (EMG) electrodes were implanted into the right masseter and digastric muscles of 6–8-week-old C57BL/6j mice. Three weeks after surgery, in vivo two-photon Ca2+ imaging of layer (L) 2/3 neurons and simultaneous EMG recordings were performed during the masticatory sequence.
Results
Mastication induced a remarkable increase in the power and frequency of Ca2+ responses and correlated with majority of the mastication-related motor cortical L2/3 neuronal activity. These mastication-related changes correlated with the activity of neurons with low baseline activity that occurred before mastication. Extraction of the right upper three molars caused clear neuroplastic changes in the mastication-induced Ca2+ activity of L2/3 neurons.
Conclusions
Our in vivo imaging study provides new insights into the neuronal basis of tooth loss-induced cortical neuroplasticity, and suggests a possible therapeutic approach for oral sensorimotor dysfunction.
{"title":"Tooth loss-associated neuroplasticity of mastication-related motor cortical neurons","authors":"Takafumi Katagiri , Shiro Nakamura , Yoshihisa Tachibana , Kiyomi Nakayama , Ayako Mochizuki , Masanori Dantsuji , Kazuyoshi Baba , Tomio Inoue","doi":"10.1016/j.job.2024.100606","DOIUrl":"10.1016/j.job.2024.100606","url":null,"abstract":"<div><h3>Objectives</h3><div>The cerebral cortex contains neurons that play a pivotal role in controlling rhythmic masticatory jaw movements. However, the population characteristics of individual cortical neuronal activity during mastication and the impact of tooth loss on these characteristics remain unclear. Thus, in this study, we aimed to determine the activity patterns of mastication-related motor cortical neurons elicited during mastication and examine the effects of tooth extraction on neuronal activity using two-photon Ca<sup>2+</sup> imaging in head-restrained awake mice.</div></div><div><h3>Methods</h3><div>GCaMP6f-expressing adeno-associated virus serotype 1 was injected into the left motor cortex (centered 2 mm anterior and 2 mm lateral to the bregma) and electromyography (EMG) electrodes were implanted into the right masseter and digastric muscles of 6–8-week-old C57BL/6j mice. Three weeks after surgery, <em>in vivo</em> two-photon Ca<sup>2+</sup> imaging of layer (L) 2/3 neurons and simultaneous EMG recordings were performed during the masticatory sequence.</div></div><div><h3>Results</h3><div>Mastication induced a remarkable increase in the power and frequency of Ca<sup>2+</sup> responses and correlated with majority of the mastication-related motor cortical L2/3 neuronal activity. These mastication-related changes correlated with the activity of neurons with low baseline activity that occurred before mastication. Extraction of the right upper three molars caused clear neuroplastic changes in the mastication-induced Ca<sup>2+</sup> activity of L2/3 neurons.</div></div><div><h3>Conclusions</h3><div>Our <em>in vivo</em> imaging study provides new insights into the neuronal basis of tooth loss-induced cortical neuroplasticity, and suggests a possible therapeutic approach for oral sensorimotor dysfunction.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 1","pages":"Article 100606"},"PeriodicalIF":2.6,"publicationDate":"2024-12-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142907728","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 : 2024-12-27DOI: 10.1016/j.job.2024.100608
Akihiro Ochiai , Atsushi Iwawaki , Yusei Otaka , Takeru Ishii , Kota Ozawa , Yuko Otomo , Shinji Kito , Hideki Saka
Objectives
This study aimed to measure the volume of the third cervical vertebra using head and neck multi-detector computed tomography (MDCT) and establish a sex determination model based on sex differences in volume.
Methods
Head and neck CT images of 85 patients were obtained for dental diagnostic and therapeutic purposes. Digital Imaging and Communications in Medicine (DICOM) data obtained from head and neck CT were constructed using a three-dimensional image analysis software. A region of interest was created that included the entire third cervical vertebra and its volume was measured. Descriptive statistics were calculated for the measurements, and the means, standard deviations, and medians of the measurements were calculated separately for men and women. To create a sex determination model, binomial logistic regression analysis was performed using the training data group, with vertebral volume and sex as the explanatory and objective variables, respectively.
Results
The mean score of men was significantly higher than that of women. The sex determination model using the volume of the third cervical vertebral body resulted in an 80% correct sex classification rate.
Conclusion
The sex determination model can be used with other regional methods to aid sex determination.
{"title":"Sex determination method using the third cervical vertebral body by head and neck CT","authors":"Akihiro Ochiai , Atsushi Iwawaki , Yusei Otaka , Takeru Ishii , Kota Ozawa , Yuko Otomo , Shinji Kito , Hideki Saka","doi":"10.1016/j.job.2024.100608","DOIUrl":"10.1016/j.job.2024.100608","url":null,"abstract":"<div><h3>Objectives</h3><div>This study aimed to measure the volume of the third cervical vertebra using head and neck multi-detector computed tomography (MDCT) and establish a sex determination model based on sex differences in volume.</div></div><div><h3>Methods</h3><div>Head and neck CT images of 85 patients were obtained for dental diagnostic and therapeutic purposes. Digital Imaging and Communications in Medicine (DICOM) data obtained from head and neck CT were constructed using a three-dimensional image analysis software. A region of interest was created that included the entire third cervical vertebra and its volume was measured. Descriptive statistics were calculated for the measurements, and the means, standard deviations, and medians of the measurements were calculated separately for men and women. To create a sex determination model, binomial logistic regression analysis was performed using the training data group, with vertebral volume and sex as the explanatory and objective variables, respectively.</div></div><div><h3>Results</h3><div>The mean score of men was significantly higher than that of women. The sex determination model using the volume of the third cervical vertebral body resulted in an 80% correct sex classification rate.</div></div><div><h3>Conclusion</h3><div>The sex determination model can be used with other regional methods to aid sex determination.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 1","pages":"Article 100608"},"PeriodicalIF":2.6,"publicationDate":"2024-12-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142903708","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}
This study aimed to elucidate the roles of Prrx1 and Prrx2, homeobox transcription factors, in tooth development and determine whether Prrx2 regulates pannexin 3 (Panx3) expression, which is important in preodontoblasts.
Methods
Tooth sections were prepared from 13.5-, 15.5-, and 18.5-day-old embryonic ICR mice, and Prrx1- and Prrx2-expressing cells were identified by in situ hybridization. To clarify the direct relationship between Prrx2 and Panx3, dual-luciferase reporter assay and electrophoretic mobility shift assay (EMSA) were performed. The effect of endogenous Prrx2 suppression on Panx3 expression was analyzed using an siRNA assay.
Results
In situ hybridization revealed that in the molars, Prrx1 and Prrx2 were similarly expressed in the bud and cap stages; however, only Prrx2 was expressed in preodontoblasts at the bell stage. In the incisors, Prrx2-expressing cells were observed from dental papilla cells to preodontoblasts. In serial sections, Prrx2-expressing cells in preodontoblasts corresponded to Panx3-expressing cells. Luciferase reporter assay using luciferase reporter plasmids containing Panx3 promoter revealed that Prrx2 overexpression in HEK293 cells significantly increased luciferase activity. EMSA of nuclear extract proteins from Prrx2-overexpressing HEK293 cells or mouse dental papilla-derived cells to the Panx3 promoter showed the protein-probe complex bands. SiRNA assay revealed that Prrx2 knockdown inhibited Panx3 expression.
Conclusions
Our results suggest that Prrx2 may regulate Panx3 expression during odontoblastic differentiation.
{"title":"Prrx2, the paired-related homeobox transcription factor, functions as a potential regulator of pannexin 3 expression in odontoblast differentiation","authors":"Manami Tanaka , Asuna Sugimoto , Kokoro Iwata , Yumiko Nakashima , Muhammad Dhiaulfikri Nauval Hadiana , Yusuke Iwabuchi , Kanae Wada , Atsushi Oishi , Tsutomu Iwamoto","doi":"10.1016/j.job.2024.100601","DOIUrl":"10.1016/j.job.2024.100601","url":null,"abstract":"<div><h3>Objectives</h3><div>This study aimed to elucidate the roles of Prrx1 and Prrx2, homeobox transcription factors, in tooth development and determine whether Prrx2 regulates pannexin 3 (Panx3) expression, which is important in preodontoblasts.</div></div><div><h3>Methods</h3><div>Tooth sections were prepared from 13.5-, 15.5-, and 18.5-day-old embryonic ICR mice, and Prrx1- and Prrx2-expressing cells were identified by <em>in situ</em> hybridization. To clarify the direct relationship between Prrx2 and Panx3, dual-luciferase reporter assay and electrophoretic mobility shift assay (EMSA) were performed. The effect of endogenous Prrx2 suppression on Panx3 expression was analyzed using an siRNA assay.</div></div><div><h3>Results</h3><div><em>In situ</em> hybridization revealed that in the molars, Prrx1 and Prrx2 were similarly expressed in the bud and cap stages; however, only Prrx2 was expressed in preodontoblasts at the bell stage. In the incisors, Prrx2-expressing cells were observed from dental papilla cells to preodontoblasts. In serial sections, Prrx2-expressing cells in preodontoblasts corresponded to Panx3-expressing cells. Luciferase reporter assay using luciferase reporter plasmids containing Panx3 promoter revealed that Prrx2 overexpression in HEK293 cells significantly increased luciferase activity. EMSA of nuclear extract proteins from Prrx2-overexpressing HEK293 cells or mouse dental papilla-derived cells to the Panx3 promoter showed the protein-probe complex bands. SiRNA assay revealed that Prrx2 knockdown inhibited Panx3 expression.</div></div><div><h3>Conclusions</h3><div>Our results suggest that Prrx2 may regulate Panx3 expression during odontoblastic differentiation.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 1","pages":"Article 100601"},"PeriodicalIF":2.6,"publicationDate":"2024-12-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142903692","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 : 2024-12-20DOI: 10.1016/j.job.2024.100603
Ai Orimoto , Ziyi Wang , Mitsuaki Ono , Chiaki Kitamura , Kentaro Ono
Objectives
Dental pulp stem cells (DPSCs) are essential for reparative dentinogenesis following damage or infection. DPSCs surrounding theblood vessels in the central region of the dental pulp actively proliferate after tooth injury and differentiate into new odontoblast-like cells or odontoblasts to form reparative dentin. However, the signaling pathways involved in undifferentiated and osteodifferentiated DPSCs under inflammatory conditions remain unclear. This study aimed to compare the expression profiles of immortalized undifferentiated and osteo-differentiated human DPSCs (hDPSCs) treated with and without lipopolysaccharide (LPS) to elucidate the molecular regulatory mechanisms involved in inflammatory conditions.
Methods
We investigated the differences between undifferentiated and osteodifferentiated hDPSCs in response to LPS. RNA-seq analyses of undifferentiated and osteodifferentiated hDPSCs were performed with and without LPS.
Results
Whole-transcriptome profiling revealed distinct differences in the expression patterns of LPS-treated undifferentiated and osteodifferentiated DPSCs. Death-associated protein kinase 1 levels downregulated in LPS-treated osteodifferentiated cells, inhibiting apoptosis and enhancing cell survival After LPS treatment, osteodifferentiated DPSCs exhibited higher expression levels of various inflammatory cytokines and chemokines than undifferentiated DPSCs.
Conclusion
This study provides valuable transcriptomic data as a critical resource for uncovering potential therapeutic targets to enhance cell survival and regulate inflammation within the dental pulp. By elucidating the key molecular mechanisms and identifying specific gene expression changes linked to inflammatory and immune responses, these findings provide significant insights into osteo-differentiated hDPSCs.
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