Journal of Oral Biosciences最新文献

{"title":"Reactivation of Epstein-Barr virus by n-butyric acid from Pseudoramibacter alactolyticus induces inflammatory cytokines in periapical granulomas.","authors":"Taiki Miyata, Osamu Takeichi, Kenichi Imai, Masayuki Okano, Seiya Inoue, Takuya Yasukawa, Yusuke Suzuki","doi":"10.1016/j.job.2024.10.001","DOIUrl":"https://doi.org/10.1016/j.job.2024.10.001","url":null,"abstract":"<p><strong>Objectives: </strong>This study investigates whether latent Epstein-Barr virus (EBV) can be reactivated by n-butyric acid from Pseudoramibacter alactolyticus, and if such reactivation induces expression of interleukin (IL)-1β and IL-6 in periapical granulomas.</p><p><strong>Methods: </strong>We analyzed periapical granulomas and healthy gingival tissues to detect the presence of EBV and P. alactolyticus. The concentration of n-butyric acid in P. alactolyticus culture supernatants was measured. BZLF-1 luciferase assays were conducted with or without these supernatants. Immunohistochemical detection of ZEBRA-, IL-1β-, and IL-6-expressing cells was performed in the tissue samples. Additionally, mRNA expression levels of BZLF-1, IL-1β, and IL-6 were quantified and statistically analyzed for correlation. The expression of these mRNAs was also measured in Daudi cells treated with or without the culture supernatants.</p><p><strong>Results: </strong>Both EBV and P. alactolyticus were detected in periapical granulomas, but not in healthy tissues. The concentration of n-butyric acid in the culture supernatants was ∼3.58 mmol/L. BZLF-1 luciferase activity in the presence of the culture supernatants was comparable to that of commercially available butyric acid, whereas no activity was detected without the supernatants. Cells expressing ZEBRA co-expressed IL-1β and IL-6. The mRNA levels of BZLF-1, IL-1β, and IL-6 in periapical granulomas were correlated with the number of EBV DNA copies. Daudi cells treated with the culture supernatants expressed BZLF-1, IL-1β, and IL-6 mRNA, while those without the supernatants did not.</p><p><strong>Conclusions: </strong>The study concludes that EBV can be reactivated by n-butyric acid produced by P. alactolyticus, leading to the induction of IL-1β and IL-6 expression in periapical granulomas.</p>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142477313","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}

{"title":"ED-71 promotes osseointegration of titanium implants in a rat model of GIOP by alleviating the effects of dexamethasone on bone remodeling in a SIRT1-dependent manner.","authors":"Chunying Li, Pengfei Xue, Guanglin Duan, Ailing Song, Runbing Zhai, Jie Ma, Minqi Li","doi":"10.1016/j.job.2024.10.003","DOIUrl":"https://doi.org/10.1016/j.job.2024.10.003","url":null,"abstract":"<p><strong>Objective: </strong>Glucocorticoid-induced osteoporosis (GIOP), a common complication of glucocorticoid usage, plays a critical role in the success of dental implant restoration by affecting osseointegration. Eldecalcitol (ED-71) prevents GIOP; however, its role in the osseointegration of implants under GIOP conditions remains elusive.</p><p><strong>Methods: </strong>Dexamethasone was used to establish a rat model of GIOP. Subsequently, mini-implant surgery was performed on the femur. GIOP rats were administered ED-71 via gavage to assess its role in the osseointegration of titanium implants under GIOP conditions. MC3T3-E1 and RAW264.7 cells were utilized to explore the molecular mechanism of ED-71 in ameliorating disorder of bone remodeling caused by dexamethasone.</p><p><strong>Results: </strong>The administration of ED-71 promoted the formation of newly formed woven bone and the resolution of inflammation around titanium implants. In vitro experiments indicated that ED-71 ameliorated dexamethasone-induced dysfunction of osteoblasts and osteoclasts by increasing the expression level of sirtuin 1 (SIRT1). Inhibition of SIRT1 by selisistat counteracts the regulatory effects of ED-71 on dexamethasone-induced disorder of bone remodeling. Molecular docking and Western blotting revealed that the neurogenic locus notch homolog protein and nuclear factor kappa B signaling pathways are essential for the effects of ED-71 on dexamethasone-induced disorder of bone remodeling.</p><p><strong>Conclusion: </strong>ED-71 promoted implant osseointegration in a rat model of GIOP by alleviating the effects of dexamethasone on bone remodeling in a SIRT1-dependent manner.</p>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142477312","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}

{"title":"Development of a new antibody drug to treat congenital tooth agenesis.","authors":"K Takahashi, H Kiso, E Mihara, J Takagi, Y Tokita, A Murashima-Suginami","doi":"10.1016/j.job.2024.10.002","DOIUrl":"https://doi.org/10.1016/j.job.2024.10.002","url":null,"abstract":"<p><strong>Background: </strong>This study aimed to develop a therapeutic agent promoting teeth regeneration from autologous tissues for congenital tooth agenesis, specifically for hypodontia (≤ 5 missing congenital teeth, 10% prevalence) and oligodontia (≥ 6 missing congenital teeth, 0.1% prevalence).</p><p><strong>Highlight: </strong>We studied mice genetically deficient in the USAG-1 protein, an antagonist of BMP/Wnt which forms excessive teeth. We identified USAG-1 as a target molecule for increasing the number of teeth. Crossing USAG-1-deficient mice with a congenital tooth agenesis model restored tooth formation. We produced anti-USAG-1 neutralizing antibodies as potential therapeutic agents for the treatment of congenital tooth agenesis. Mice anti-USAG-1 neutralizing antibodies can potentially rescue the developmentally arrested tooth germ programmed to a certain tooth type. A humanized anti-USAG-1 antibody was developed as the final candidate.</p><p><strong>Conclusion: </strong>Targeting USAG-1 shows promise for treating missing congenital tooth. Anti-USAG-1 neutralizing antibodies have been developed and will progress towards clinical trials, which may regenerate missing congenital teeth in conditions, such as hypodontia and oligodontia. The protocol framework for a phase 1 study has been finalized, and preparation for future studies is underway.</p>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142401587","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}

{"title":"Herbal medicine Ninjinyoeito inhibits RANKL-induced osteoclast differentiation and bone resorption activity by regulating NF-kB and MAPK pathway.","authors":"Kaung Htike, Kunihiro Yoshida, Takanori Eguchi, Katsuki Takebe, Xueming Li, Yaxin Qu, Eiko Sakai, Takayuki Tsukuba, Kuniaki Okamoto","doi":"10.1016/j.job.2024.09.007","DOIUrl":"10.1016/j.job.2024.09.007","url":null,"abstract":"<p><strong>Objectives: </strong>Osteoporosis is a systemic bone metabolism disorder characterized by decreased bone mass and strength. Osteoclasts (OCs) are giant multinucleated cells that regulate bone homeostasis by degrading bone matrix. Excessive OC differentiation and activity can lead to serious bone metabolic disorders including osteoporosis. Current treatments, including antiresorptive drugs, exert considerable adverse effects, including jaw osteonecrosis. Herbal medicines, such as Ninjinyoeito (NYT), may also offer efficacy, but with fewer adverse effects. In this study, we investigated NYT's effects on osteoclastogenesis.</p><p><strong>Methods: </strong>Tartrate-resistant acid phosphatase (TRAP) staining and bone resorption assays were performed to examine NYT's effects on OC differentiation and function. OC-related gene expression at mRNA and protein levels was investigated to confirm NYT's inhibitory action against osteoclastogenesis. We also demonstrated involvement of signaling pathways mediated by IκBα and mitogen-activated protein kinases (MAPK) [extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38] and showed nuclear translocation of nuclear factor of activated T-cell cytoplasmic 1 (NFATc1) and nuclear factor kappa B (NF-κB) p65 during osteoclastogenesis.</p><p><strong>Results: </strong>TRAP staining and bone resorption assays confirmed that NYT significantly inhibited OC differentiation and function. Western blot and RT-PCR results showed that NYT ameliorated osteoclastogenesis by suppressing mRNA and protein level expression of OC-related genes. Moreover, blots and immunocytochemistry (ICC) data clarified that NYT abrogates signaling pathways mediated by IκBα and MAPK (ERK, JNK, p38), and demonstrated nuclear translocation of NFATc1 and NF-κB p65 during OC differentiation.</p><p><strong>Conclusions: </strong>These findings suggest NYT is an alternative therapeutic candidate for treating osteoporosis.</p>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142376160","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}

{"title":"Comparison of rhythmic jaw muscle activities induced by electrical stimulations of the corticobulbar tract during rapid eye movement sleep with those during wakefulness and non-rapid eye movement sleep in freely moving guinea pigs.","authors":"Makoto Higashiyama, Yuji Masuda, Ayano Katagiri, Hiroki Toyoda, Masaharu Yamada, Atsushi Yoshida, Takafumi Kato","doi":"10.1016/j.job.2024.09.004","DOIUrl":"https://doi.org/10.1016/j.job.2024.09.004","url":null,"abstract":"<p><strong>Objective: </strong>Rhythmic jaw muscle activities (RJMAs) occur during rapid eye movement (REM) sleep in humans and animals even though motoneurons are inhibited. The present study compared the characteristics of jaw muscle activities induced by electrical microstimulations of the corticobulbar tract (CT) during REM sleep with those during wakefulness and non-REM sleep.</p><p><strong>Methods: </strong>Eleven guinea pigs were surgically prepared for polygraphic recordings with the implantation of a stimulating electrode. Long- and short-train repetitive electrical microstimulations were applied to the CT under freely moving conditions. The response rate, latency, burst amplitude, and cycle length in the digastric muscle were calculated and cortical and cardiac activities were quantified.</p><p><strong>Results: </strong>Long-train microstimulations induced RJMAs in the digastric muscle followed by masseter muscle activity during wakefulness and non-REM sleep and only induced rhythmic digastric muscle activity during REM sleep. The response rate of RJMAs and the burst amplitude of digastric muscles were significantly lower during REM sleep than during wakefulness and non-REM sleep. However, response latency did not significantly differ between REM sleep and wakefulness. Transient cortical and cardiac changes were associated with RJMAs induced during non-REM sleep, but not during REM sleep. Short-train microstimulations induced a short-latency digastric response, the amplitude of which was significantly lower during REM sleep than during non-REM sleep and wakefulness.</p><p><strong>Conclusions: </strong>These results suggest that the masticatory CPG was activated by electrical CT stimulations independently of the motoneuron inhibitory system during REM sleep.</p>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142298230","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}

{"title":"Potassium nitrate suppresses hyperactivities of Vc neurons of the model with dentin hypersensitivity.","authors":"Shiori Sugawara, Koichi Iwata, Toshiki Takamizawa, Masashi Miyazaki, Masayuki Kobayashi","doi":"10.1016/j.job.2024.09.005","DOIUrl":"https://doi.org/10.1016/j.job.2024.09.005","url":null,"abstract":"<p><strong>Objective: </strong>Potassium nitrate (KNO₃) suppresses nociception induced by dental hypersensitivity (HYS). We aimed to examine the effects of KNO₃ on the neural activity of the trigeminal spinal subnucleus caudalis (Vc) in HYS model rats.</p><p><strong>Methods: </strong>KNO₃ or vehicle was applied to the exposed dentin of HYS rats for 3 days. c-Fos expression and neuronal activity in the Vc after acetone treatment for cold stimulation were examined to evaluate the effects of KNO₃ application on dentin.</p><p><strong>Results: </strong>The number of c-Fos-immunoreactive cells in the Vc was lower in the group that received KNO₃ (KNO₃ group) than in the group that received vehicle (control group). Spike firing of Vc neurons in response to cold stimulation of the dentin was recorded before and after KNO₃ application to the cavity, and the increased neural activity was effectively suppressed by KNO₃ application. Scanning electron microscopy revealed that the dentin tubules were not occluded by deposits in any of the groups.</p><p><strong>Conclusions: </strong>KNO₃-induced suppression of Vc neuronal activity does not involve physical occlusion of the dentin tubules but likely involves suppression of Aδ or C-fiber activities in the tooth pulp, resulting in the suppression of Vc neuronal activities.</p>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142298231","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}

{"title":"Antihypertensive agent losartan promotes tongue squamous cell carcinoma cell proliferation via EGFR/ERK1/2/cyclin D1 signaling axis.","authors":"Luo-Yun Wu, Bor-Chyuan Su, Hsin-Hsien Yu, Chih-Cheng Cheng, Chia-Chi Tsai, Pei-Ling Hsu, Chu-Wan Lee","doi":"10.1016/j.job.2024.09.003","DOIUrl":"https://doi.org/10.1016/j.job.2024.09.003","url":null,"abstract":"<p><strong>Objective: </strong>To study the effects of losartan, an angiotensin II receptor blocker, in the SCC4 and SCC25 human tongue squamous cell carcinoma cell lines.</p><p><strong>Methods: </strong>Cell proliferation was measured by MTS/PMS activity and trypan blue exclusion assays. The levels of the cell proliferation marker, cyclin D1, were analyzed by western blotting. Apoptosis was assessed by caspase-3 activation and Annexin V-FITC/propidium iodide double staining. Activation of epidermal growth factor receptor (EGFR) and ERK1/2 was validated by western blotting.</p><p><strong>Results: </strong>Moderate concentrations of losartan enhanced the proliferation of SCC4 and SCC25 cells. However, high losartan concentrations induced apoptosis in SCC4 cells. Losartan activated the EGFR/ERK1/2/cyclin D1 signaling axis, which in turn promoted cell proliferation. Afatinib (EGFR inhibitor) and U0126 (ERK1/2 inhibitor) abolished losartan-induced cell proliferation. In contrast, UC2288 (p21 inhibitor) enhanced it.</p><p><strong>Conclusions: </strong>Losartan exhibited dual effects on tongue squamous cell carcinoma cells. Moderate losartan concentrations facilitated cell proliferation, whereas high concentrations induced cytotoxicity in tongue carcinoma cells.</p>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142157193","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}

{"title":"Productions of Th2 cytokines, IL-4 and IL-10, were enhanced via the function of IL-2 from anti-CD3 antibody-stimulated mouse spleen cells treated with caffeic acid phenethyl ester.","authors":"Moe Takahashi, Masako Mizuno-Kamiya, Shifa Rahman, Hanemi Tsuruta, Kumiko Ikeno, Harumi Kawaki, Genjiro Nakamura, Yasunori Muramatsu, Toru Nikaido, Hisakazu Fujita, Nobuo Kondoh","doi":"10.1016/j.job.2024.09.001","DOIUrl":"https://doi.org/10.1016/j.job.2024.09.001","url":null,"abstract":"<p><strong>Objectives: </strong>Interleukin (IL)-2 production by mouse spleen cells stimulated with an anti-CD3 antibody is significantly enhanced by caffeic acid phenethyl ester (CAPE), a major constituent of Chinese propolis (CP). In this study, we evaluated the functional significance of IL-2 in CAPE-treated activated spleen cells.</p><p><strong>Methods: </strong>Mouse spleen cells were stimulated with an anti-CD3 monoclonal antibody in the presence of CAPE. Cytokine production was examined using an enzyme-linked immunosorbent assay (ELISA). Messenger RNA level expression was examined via reverse transcription quantitative polymerase chain reaction (RT-PCR). IL-2 function was assessed using IL-2 and a neutralizing antibody. Spleen cell subsets were identified and characterized using flow cytometry.</p><p><strong>Results: </strong>CAPE treatment of anti-CD3 antibody-stimulated spleen cells reduced IFN-γ production, then enhanced IL-2 production, followed by enhancement of IL-4 and IL-10 production. The Th2 cytokine production enhancing effects of CAPE were completely abolished by addition of an anti-IL-2 neutralizing antibody. In the absence of CAPE, exogenously added IL-2 could enhance IL-4 production to a lesser degree, but did not stimulate IL-10 production, in stimulated spleen cells. Interestingly, CAPE significantly reduced the proportions of CD4⁺ and CD8⁺ cells, and increased those of CD4^-CD8^- cells among anti-CD3 stimulated spleen cells, in the presence or absence of anti-IL-2 neutralizing antibody treatment.</p><p><strong>Conclusions: </strong>CAPE reduced IFN-γ production, then enhanced IL-4 and IL-10 production via the activity of specifically elevated IL-2 in stimulated spleen cells. CAPE exerted these effects in a CD4^- CD8^- cell specific manner.</p>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142146567","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}

{"title":"STAT3 interactome predicts presence of proteins that regulates immune system in oral squamous cell carcinoma.","authors":"Rajdeep Chakraborty, Pallavi Khodlan, Aidan Tay, Fei Liu","doi":"10.1016/j.job.2024.09.002","DOIUrl":"https://doi.org/10.1016/j.job.2024.09.002","url":null,"abstract":"<p><strong>Objectives: </strong>Signal transducer and activator of transcription 3 (STAT3) is one of the key proliferation mechanism-related proteins that helps in oral squamous cell carcinoma (OSCC) progression. Immune evasion by STAT3 is mediated by the JAK2/STAT3/PDL1 signaling axis. Based on previous findings, we hypothesized that STAT3-binding partners participate in the inhibition of anti-tumor activity in OSCC.</p><p><strong>Methods: </strong>A 3D cancer-immune co-culture model was constructed using oral cancer cell lines SCC4, SCC9, SCC25, and CAL27 and normal oral cell line OKF6. The cells were co-cultured with natural killer (NK-92) and Jurkat cells. The target protein STAT3 was chosen based on SWATH data, and co-immunoprecipitation (Co-IP)-based proteomics was conducted. The Co-IP LC-MS/MS output was analyzed to determine the protein interaction network, gene ontology, pathway analysis, and protein cluster annotation.</p><p><strong>Results: </strong>STAT3 in oral cancer cell lines interacts with the epidermal growth factor receptor (EGFR) and other proteins that participate in proliferation and immune mechanisms. Proteome analysis showed that some STAT3-binding proteins found in this study are known immune system regulators.</p><p><strong>Conclusion: </strong>Overall, STAT3 interactive proteins regulate the immune system in oral squamous cell carcinoma cells.</p>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142141360","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}

{"title":"Y-27632 enables long-term expansion of mouse submandibular gland epithelial cells via inactivation of TGF-β1/CTGF/p38 and ROCK2/JNK signaling pathway.","authors":"Kichul Kim, Naeun Oh, Hyewon Kim, Sangho Roh","doi":"10.1016/j.job.2024.08.005","DOIUrl":"https://doi.org/10.1016/j.job.2024.08.005","url":null,"abstract":"<p><strong>Objectives: </strong>This study aimed to investigate the effects of Y-27632 on the long-term maintainence of mouse submandibular epithelial cells (SG-Epis) in vitro and to elucidate the underlying mechanisms.</p><p><strong>Methods: </strong>The role of the Rho-associated kinase (ROCK) inhibitor Y-27632 in maintaining SG-Epis and its underlying mechanisms were evaluated by examining the in vitro expansion of mouse SG-Epis. Changes in key cellular characteristics, such as proliferation, long-term expansion, and mRNA and protein expression, were assessed in the presence or absence of Y-27632.</p><p><strong>Results: </strong>Treatment with Y-27632 significantly enhanced the proliferative potential of SG-Epis, preserving Krt8 and Krt14 expression over 17 passages. In the absence of Y-27632, SG-Epis lost their epithelial morphology. However, Y-27632 treatment maintained the epithelial morphology and downregulated mRNA levels of Tgf-β1, Ctgf, and Rock2. Treatment with TGF-β1 indicated that TGF-β/CTGF/p38 signaling is responsible for the maintenance of SG-Epis, while RNA interference studies revealed that ROCK2/c-Jun N-terminal kinase (JNK) signaling is also crucial for SG-Epis proliferation and maintenance.</p><p><strong>Conclusions: </strong>The TGF-β1/CTGF/p38 and ROCK2/JNK signaling pathways are responsible for SG-Epis proliferation, and Y-27632 treatment effectively inactivates these pathways, enabling long-term in vitro maintenance of SG-Epis. The culture method utilizing Y-27632 provides an effective approach for the in vitro expansion of SG-Epis.</p>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142120851","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}