Journal of Dental Research最新文献

{"title":"DHFR-Driven Metabolic Memory Sustains Periodontal Tissue Destruction","authors":"L. Nie, Y. Sun, H. Dong, M. You, A. Cui, Z. Yue, P. Zhao, Q. Lv, N. Ji, H. Wang, X. Xu, W.K. Leung, J. Wang, Q. Wang","doi":"10.1177/00220345251340632","DOIUrl":"https://doi.org/10.1177/00220345251340632","url":null,"abstract":"Diabetes mellitus is one of the most common metabolic diseases worldwide, with periodontal tissue destruction being a major complication. Hyperglycemia-induced changes in metabolism and immune responses may lead to persistent periodontal tissue destruction. This study aimed to investigate hyperglycemia-induced chronic periodontal tissue destruction by focusing on dihydrofolate reductase (DHFR) and its role in metabolic memory. We used CD45.2 <jats:sup>+</jats:sup> BKS-Lepr <jats:sup>em2Cd479</jats:sup> /Gpt mice and CD45.1 <jats:sup>+</jats:sup> FVB/NJGpt mice to construct metabolic memory and bone marrow transplantation models, respectively. Our findings showed that hyperglycemia induced a persistent inflammatory senescent phenotype in macrophages. Insulin glycemic control was unable to reverse these pathological changes in bone marrow–derived macrophages and gingival tissues. Furthermore, combined metabolomic and transcriptomic analyses revealed reduced DHFR-mediated 1-carbon metabolism under hyperglycemia, with decreased levels of the reduced form of nicotinamide-adenine dinucleotide phosphate and adenosine triphosphate caused by altered glucose metabolism, impairing the function of DHFR. Alterations in DNA methylation may be responsible for memory-like metabolic patterns in macrophages. Finally, DHFR overexpression reversed hyperglycemia-induced persistent metabolic and pathological changes in macrophages. In summary, this study highlights DHFR-mediated metabolic memory in macrophages as a key factor driving hyperglycemia-induced chronic periodontal tissue destruction.","PeriodicalId":15596,"journal":{"name":"Journal of Dental Research","volume":"10 1","pages":""},"PeriodicalIF":7.6,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144500629","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cancer-Associated Fibroblast-Driven Progression of HPV+ Head and Neck Cancer with Organoid Models","authors":"P.X. Xue, R.R. Xiao, L. Chen, G.Z. Yang, C.T. Fan, J.Y. Zhu, J.C. Zhao, H. Zhang, L.L. Sun, S.Y. Sun","doi":"10.1177/00220345251342168","DOIUrl":"https://doi.org/10.1177/00220345251342168","url":null,"abstract":"Human papillomavirus–positive head and neck cancer (HPV+ HNC) is a distinct tumor entity that differs significantly from HPV-negative HNC (HPV− HNC) in clinical characteristics, genetic variations, and biological behavior. Focusing on the disease-specific features of HPV+ HNC is crucial for understanding its mechanisms of occurrence and progression as well as for advancing therapeutic strategies. Compared with HPV− cases, we found that HPV+ HNCs were enriched in cancer-associated fibroblasts (CAFs), and CAFs contribute significantly to the poor prognosis of HPV+ HNCs. To better explore the interaction between HPV+ HNCs and CAFs, we pioneered the development of 5 HPV+ HNC patient-derived organoid (PDO) models, which faithfully recapitulate the HPV infection status and the distinct drug responses of HPV+ HNCs. Using the generated HPV+ PDOs, we discovered that CAFs significantly enhanced HPV16 E7 expression in HPV+ cases while also promoting proliferation, stemness, and drug resistance. Moreover, our data showed that interleukin-6 (IL6), secreted by CAFs, regulates the activation of the hypoxia-inducible factor 1A (HIF1A) pathway in HPV+ cases. Drugs targeting HIF1A and the IL6 receptor significantly reduce HPV16 E7 expression and the proliferation of HPV+ HNC PDOs. These findings uncover unique cellular and molecular factors that drive the malignant progression of HPV+ HNC and hold promise for the development and application of novel therapeutic strategies for this disease.","PeriodicalId":15596,"journal":{"name":"Journal of Dental Research","volume":"246 1","pages":""},"PeriodicalIF":7.6,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144500677","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Aggregatibacter aphrophilus T6SS Effectors in Host–Bacterial Interactions","authors":"K. Bao, J. Oscarsson, P. Gehring, J. Grossmann, G.N. Belibasakis, N. Bostanci","doi":"10.1177/00220345251337745","DOIUrl":"https://doi.org/10.1177/00220345251337745","url":null,"abstract":"<jats:italic>Aggregatibacter aphrophilus</jats:italic> is the only known oral bacterium with a functional type VI secretion system (T6SS) that acts against <jats:italic>Aggregatibacter actinomycetemcomitans</jats:italic> . Bacteria use the T6SS to deliver toxic effectors into bacterial or eukaryotic cells during interbacterial competition or host colonization. To date, the T6SS of <jats:italic>A. aphrophilus</jats:italic> has not been demonstrated to participate in antieukaryotic activity, nor have the cytotoxic effectors involved been identified. Here, we identified 2 T6SS effectors in <jats:italic>A. aphrophilus</jats:italic> , a glycosyl hydrolase (Glh) and a phospholipase D (Tle5), which, upon inactivation of their respective genes, together resulted in abolished T6SS activity against <jats:italic>A. actinomycetemcomitans</jats:italic> in multispecies biofilms. Next, we probed the role of the 2 T6SS effectors in host–cell interactions using gingival keratinocytes. Interestingly, although neither of the effectors appeared to contribute to the acute inflammatory response directly, the co-presence of both species reduced the inflammatory effect, likely due to the T6SS-dependent elimination of <jats:italic>A. actinomycetemcomitans</jats:italic> , hence decreasing the bacterial abundance. This reduction was not observed using <jats:italic>A. aphrophilus</jats:italic> mutants lacking the effectors or the T6SS “tube” core protein, hemolysin co-regulated protein (Hcp). Here, we show that the T6SS effectors in <jats:italic>A. aphrophilus</jats:italic> have distinct functions in eukaryotic versus bacterial cell interactions. Hence, these T6SS effectors may represent novel mechanisms of interaction between bacteria and the oral–mucosal barrier, offering potential therapeutic targets for managing periodontal pathogens.","PeriodicalId":15596,"journal":{"name":"Journal of Dental Research","volume":"17 1","pages":""},"PeriodicalIF":7.6,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144500811","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Targeting the IL-6–Th17–Neutrophil Axis Reduces Local Inflammation in Stomatitis","authors":"Z. Wang, Y. Wang, D. Zhang, Y. Lin, X. Fan, J. Liu, W. Zhang, H. Xu, N. Ji, B. Huang, Q. Chen","doi":"10.1177/00220345251338031","DOIUrl":"https://doi.org/10.1177/00220345251338031","url":null,"abstract":"Recurrent aphthous stomatitis (RAS) is a common chronic oral disease with unclear pathogenesis. Chronic inflammation associated with RAS has been linked to the dysregulation of local immune responses. We used a murine model of acetic acid–induced chemical stomatitis (CS) to assess changes in the systemic and local immune environments of CS mice relative to healthy mice. Flow cytometry revealed a significant increase in neutrophil infiltration and elevated proportions of Th1 (CD4 <jats:sup>+</jats:sup> IFN-γ <jats:sup>+</jats:sup> ) and Th17 (CD4 <jats:sup>+</jats:sup> IL-17a <jats:sup>+</jats:sup> ) cells in the lingual mucosa of CS mice 7 d after CS induction, indicating an active inflammatory response in the CS immune microenvironment. Given that Th17 cells indirectly recruit neutrophils, we used <jats:italic>Rorc</jats:italic> <jats:sup>-/-</jats:sup> mice to evaluate the effects of Th17 cell depletion. Neutrophil infiltration was markedly reduced, and decreased tissue damage was observed in the lingual mucosa of <jats:italic>Rorc</jats:italic> <jats:sup>-/-</jats:sup> mice, as confirmed by hematoxylin and eosin staining. To further investigate the mechanism underlying Th17 cell generation in stomatitis, we induced CS in <jats:italic>Il6ra</jats:italic> <jats:sup>-/-</jats:sup> mice, which exhibited significantly reduced inflammatory cell infiltration and ulcer severity in the lingual mucosa. Treatment with an anti-Ly6G antibody treatment, which can directly target and deplete neutrophils, also significantly reduced local inflammation in the CS mouse immune microenvironment and diminished Th1 and Th17 cell infiltration, indicating a positive feedback loop between Th17 cells and neutrophils in stomatitis. In conclusion, the IL-6–Th17–neutrophil axis plays a critical role in stomatitis pathogenesis, suggesting that targeting this axis could present a novel immunotherapeutic strategy for alleviating mucosal damage in patients with RAS.","PeriodicalId":15596,"journal":{"name":"Journal of Dental Research","volume":"18 1","pages":""},"PeriodicalIF":7.6,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144500666","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Birth Cohort Study Identifies Candida albicans as a Risk Factor for Dental Caries","authors":"N. Alkhars, S. Manning, N. Al Jallad, Y. Zeng, T.T. Wu, C. Fogarty, M. Mendoza, E. van Wijngaarden, D.T. Kopycka-Kedzierawski, R. Billings, K. Fiscella, H. Koo, J. Xiao","doi":"10.1177/00220345251340040","DOIUrl":"https://doi.org/10.1177/00220345251340040","url":null,"abstract":"<jats:italic>Candida albicans</jats:italic> has been implicated as a potential cariogenic microorganism, yet no prospective longitudinal studies have examined its role in severe early childhood caries (S-ECC). This study aimed to evaluate the association between oral <jats:italic>C. albicans</jats:italic> and the onset of S-ECC in a longitudinal setting. This prospective birth cohort study (2018 to 2023) enrolled 186 low-income pregnant women in their third trimester in Western New York, United States. Overall, 160 eligible infants born to these women were followed from birth to 2 y at 7 time points. Oral samples were collected to assess <jats:italic>Candida</jats:italic> species ( <jats:italic>C. albicans</jats:italic> , <jats:italic>Candida krusei</jats:italic> , and <jats:italic>Candida glabrata</jats:italic> ) and <jats:italic>Streptococcus mutans</jats:italic> . The primary outcome was the onset of S-ECC. Two-step LASSO (least absolute shrinkage and selection operator)–penalized logistic regression models were developed to identify predictive factors for S-ECC from 234 covariates grouped by their proximal association with S-ECC: infant oral microorganisms, biological-environmental factors, and maternal characteristics. Logistic regression was used to validate the association between <jats:italic>C. albicans</jats:italic> and S-ECC. Among the 118 children who completed the study, 28% developed S-ECC. The racial background of the cohort was 57% Black, 21% White, and 22% other. Oral <jats:italic>C. albicans</jats:italic> colonized in 12% of infants at 1 wk, peaking at 57% by 18 mo. Salivary <jats:italic>C. albicans</jats:italic> was associated with a 4.47-fold increased risk for S-ECC (odds ratio [OR]; 95% CI, 1.28 to 15.58; <jats:italic>P</jats:italic> = 0.02), in addition to other risk factors, including plaque score (OR, 5.19; 95% CI, 2.10 to 12.83) and salivary <jats:italic>S. mutans</jats:italic> (OR, 9.74; 95% CI, 2.27 to 41.79). <jats:italic>C. albicans</jats:italic> demonstrated strong time sensitivity in predicting S-ECC as early as 1 mo, contrasting with <jats:italic>S. mutans</jats:italic> , which exhibited predictive ability after 1 y. Oral <jats:italic>C. albicans</jats:italic> could serve as a novel biological marker for predicting ECC risk in infancy, shining a light on opportunities to develop innovative caries-predictive and preventive strategies targeting fungal contributions in pediatric care settings.","PeriodicalId":15596,"journal":{"name":"Journal of Dental Research","volume":"90 1","pages":""},"PeriodicalIF":7.6,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144503574","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Single-Cell Insights into Unicystic and Solid Ameloblastoma Heterogeneity","authors":"C. Feng, X. Dou, S.-R. Li, X. Xu, Z. Dang, Z. Jiang, E. Jiang, Z. Shang","doi":"10.1177/00220345251340892","DOIUrl":"https://doi.org/10.1177/00220345251340892","url":null,"abstract":"The clinical classification of ameloblastoma (AM) plays a decisive role in the selection of treatment options, but the difference of single-cell landscape among clinical classifications is still unclear. At the same time, there is an urgent need to understand the key cell subtypes that determine the clinical subtypes. We characterized the single-cell transcriptional profiles of clinical subtypes of AM. We also characterized a pseudotime transition trajectory from immunoactive epithelial cells to vascular-associated fibroblasts, identifying key transcription factors involved in this process. Notably, we observed significant heterogeneity between M1 and M2 macrophages among the clinical subtypes of AM. Furthermore, our analysis revealed that metabolic disorder in AM was primarily driven by the metabolic disturbances in M1 and M2 macrophages. At the cellular communication level, we highlighted the role of M2 macrophages in mediating cell interactions, focusing on the RANKL/RANK pathway associated with osteoclast activity. Finally, we attempted to establish a unicystic AM–derived epithelial cell line and utilized it to construct an AM-like organoid model; we found that M2 macrophages competed with AM for L-cysteine to achieve cystic changes in the solid lesion. Our exploration of pathogenesis underlying various clinical types of AM advances our knowledge of AM heterogeneity, offering promising targets for novel therapeutic strategies.","PeriodicalId":15596,"journal":{"name":"Journal of Dental Research","volume":"10 1","pages":""},"PeriodicalIF":7.6,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144503578","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Axin2 Deficiency Causes Hypomineralization and Delayed Tooth Development","authors":"J. Song, B. Kim, J. Na, J.J. Ryu, H.-C. Park, J.S. Shim","doi":"10.1177/00220345251342967","DOIUrl":"https://doi.org/10.1177/00220345251342967","url":null,"abstract":"Axin2 negatively regulates the Wnt signaling pathway, which plays a crucial role in multiple tooth development phases. This study aimed to investigate the effects of Axin2 deficiency on tooth development in zebrafish, which are categorized as polyphyodonts. To investigate the role of Axin2 in tooth formation, we generated Axin2 knockout zebrafish using the CRISPR/Cas9 system. Tooth development in zebrafish was observed in the transgenic line, which specifically emits fluorescence in dental tissues. Micro–computed tomography and field emission scanning electron microscopy were used to evaluate tooth shape. Tooth elemental composition was examined using energy-dispersive spectrometry analysis, and gene expression was determined using quantitative polymerase chain reaction. The results showed that the body length of the Axin2 mutant was significantly lower than that of the wild type. β-Catenin expression increased in the dental pulp of the Axin2 mutant compared with that of Axin2 <jats:sup>+/+</jats:sup> zebrafish. Confocal imaging showed that initiation and morphogenesis of teeth were sequentially delayed at positions 2V and 1V. Dentin elements, including phosphorus, calcium, carbon, and oxygen, were detected at significantly low levels in Axin2 <jats:sup>−/−</jats:sup> zebrafish. In addition, <jats:italic>runx2a</jats:italic> , <jats:italic>runx2b, sp7, scpp5</jats:italic> , and <jats:italic>col1a1a/b</jats:italic> expression was significantly reduced in Axin2 <jats:sup>−/−</jats:sup> . The results show that the loss of Axin2 function causes hypomineralization and delayed tooth development in zebrafish and suggest that spatiotemporal regulation of Wnt signaling is necessary for the normal maturation of odontoblasts.","PeriodicalId":15596,"journal":{"name":"Journal of Dental Research","volume":"17 1","pages":""},"PeriodicalIF":7.6,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144500477","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Vascular Invasion of the Dental Epithelium Is Essential for Ameloblasts","authors":"H. Asrar, J.M. Fons, A.S. Tucker","doi":"10.1177/00220345251341850","DOIUrl":"https://doi.org/10.1177/00220345251341850","url":null,"abstract":"The tooth is a highly vascularized organ. During odontogenesis, blood vessels enter the forming tooth through the dental papilla and surround the dental epithelium from the cap stage. We show that during the late bell stage, endothelial cells invade the outer enamel epithelium (OEE) and migrate through the stellate reticulum to vascularize the forming ameloblast layer. This migration was evident in both mouse and human tooth germs and is likely to represent a conserved mechanism. Migration was coordinated by dynamic changes in <jats:italic>Vegf</jats:italic> expression in the dental epithelium, with expression at the OEE at the bell stage shifting toward the ameloblast layer. Invasion through the OEE involved loss of integrity of the basement membrane, downregulation of tight junctions, and apoptosis of some OEE cells. Changes in the OEE were dependent on Hedgehog signaling, with a failure to invade in <jats:italic>K14creSmoothenedfl/fl</jats:italic> mice, where epithelial cells cannot respond to Hedgehog signaling. The impact of failed migration through the OEE was followed in <jats:italic> Cdh5creERT2 Vegfr2 <jats:sup>fl/fl</jats:sup> </jats:italic> mutant mice, where the endothelial cells cannot respond to vascular endothelial growth factor (VEGF). Failure of OEE invasion resulted in differentiation defects and extensive cell death of the ameloblast layer, highlighting the essential requirement for vascularization for development of this layer. Our results reveal the essential role of Hedgehog and VEGF signaling in correct vascularization of the tooth germ epithelial layers, allowing breakdown of the OEE and targeting endothelial cell migration into the epithelium. A better understanding of the molecular regulation of endothelial cells will help decipher how this cell population interacts with different cells of the enamel organ and will aid in attempts to revascularize teeth.","PeriodicalId":15596,"journal":{"name":"Journal of Dental Research","volume":"57 1","pages":""},"PeriodicalIF":7.6,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144500468","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Nano-Toothbrush for Noninvasive Control of Periodontitis","authors":"X. Liu, Y. Liang, Z. Li, C. Xie, Y. Zhang, W. Wang, S. Ge, J. Li","doi":"10.1177/00220345251335922","DOIUrl":"https://doi.org/10.1177/00220345251335922","url":null,"abstract":"Periodontitis is a prevalent oral disease, and current clinical management remains limited to mechanical debridement of subgingival plaque and adjunctive antibiotic therapy. In this study, we introduced a piezocatalytic nanoparticle-constructed “nano-toothbrush” as a noninvasive strategy for oral biofilm removal. The dextran-mediated biofilm affinity of the nano-toothbrush induced localized oxidative stress under ultrasound irradiation, effectively disrupting biofilms and inhibiting pathogenic bacteria. This efficacy was evidenced by the removal of ex vivo biofilms from periodontitis patients. Importantly, the selective action of the nano-toothbrush minimized the damage to mouse fibroblasts and human immortalized oral epithelial cells, affirming its biosafety. In vivo studies showed that the nano-toothbrush significantly inhibited the cell viability of <jats:italic>Porphyromonas gingivalis</jats:italic> , effectively suppressed alveolar bone resorption, and alleviated inflammatory responses in a rat periodontitis model while concurrently preserving the diversity of the oral microbiome. This study presents a promising noninvasive and precise method for the removal of oral biofilms, offering new perspectives on the application of nanotherapeutics in the treatment of periodontitis and other plaque-induced diseases.","PeriodicalId":15596,"journal":{"name":"Journal of Dental Research","volume":"31 1","pages":""},"PeriodicalIF":7.6,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144500628","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Single-Cell Atlas of Immune Microenvironment in Orthodontic Tooth Movement","authors":"J. Wang, C. Tao, H. Liu, Y. Yang, Y. Zhao, K. Tan, F. Chen, E. Yang, Y. Huang, W. Li","doi":"10.1177/00220345251334583","DOIUrl":"https://doi.org/10.1177/00220345251334583","url":null,"abstract":"Orthodontic treatment depends on periodontal tissue remodeling. While the immune system has been found to regulate this process, previous studies mostly focused on the static molecular changes in conventional immune cell sets. A recent study used single-cell RNA sequencing (scRNA-seq) to analyze macrophages under orthodontic tooth movement (OTM), introducing new subclusters and functions. However, changes in other immune cells and their interactions with surrounding stromal cells are yet unclarified. Therefore, we performed scRNA-seq in mice, aiming to describe a more comprehensive immune landscape during OTM, while further exploring the dynamic changes, functions, and cellular interactions of immune cells at a higher resolution. We first confirmed the dynamic activation of osteoclasts, osteoblasts, and immune cells at various time points. The scRNA-seq analysis identified 7 cell lineages and 18 major cell types, with immune cells forming the largest proportion. Monocytic cells, granulocytes, and lymphocytes were selected for individual reclustering, followed by analysis of specific gene expression, functional enrichment, and distribution changes during OTM. Pseudotime analysis was applied to monocytic cells and granulocytes. We identified 4 developmental pathways in monocytic cells toward dendritic cells, different subsets of macrophages, and osteoclasts. Monocytic cells tended to be more differentiated during OTM. Meanwhile, in granulocytes, neutrophil subclusters were all highly differentiated. Additionally, we assessed the cellular interactions during OTM, revealing enhanced signaling from macrophages toward osteoclasts, especially in Ccl, Tnf, and Spp1 pathways. We identified that the <jats:italic> C3ar1 <jats:sup>+</jats:sup> </jats:italic> macrophage subcluster expressed these cytokines at a high level and was enriched in positive regulation of the mitogen-activated protein kinase cascade, indicating its positive regulation toward osteoclast activity. In conclusion, this study revealed the complex immune microenvironment during OTM, providing a detailed perspective on the diverse immune cell types, their specific functions, and cellular interactions.","PeriodicalId":15596,"journal":{"name":"Journal of Dental Research","volume":"13 1","pages":""},"PeriodicalIF":7.6,"publicationDate":"2025-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144500679","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}