{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T04:16:26Z","timestamp":1790223386948,"version":"4.1.0"},"reference-count":53,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,1,6]],"date-time":"2026-01-06T00:00:00Z","timestamp":1767657600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100017686","name":"Fujian Provincial Health Technology Project","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100017686","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Bioeng. Biotechnol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>Periodontal ligament stem cells (PDLSCs) represent a promising therapeutic cell source for regenerating periodontal tissues impaired by disease. While exogenous BMP4 has been used to induce human PDLSC differentiation into distinct periodontal cell lineages, its dose-dependent effects on lineage specification remain poorly understood.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Here, we systematically investigated how BMP4 dosage modulates human PDLSC differentiation toward osteogenic and tenogenic lineages using single-cell RNA sequencing and\n                      <jats:italic>in vitro<\/jats:italic>\n                      functional assays.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Single-cell transcriptomic analysis revealed that endogenous\n                      <jats:italic>BMP4<\/jats:italic>\n                      expression inversely correlated with\n                      <jats:italic>RUNX2<\/jats:italic>\n                      but positively correlated with\n                      <jats:italic>SCX<\/jats:italic>\n                      within PDLSC subpopulations. Pseudotemporal analysis demonstrated biphasic differentiation dynamics: low\n                      <jats:italic>BMP4<\/jats:italic>\n                      expression corresponded to high expression of both markers, while\n                      <jats:italic>BMP4<\/jats:italic>\n                      upregulation inversely correlated with\n                      <jats:italic>RUNX2<\/jats:italic>\n                      but positively correlated with\n                      <jats:italic>SCX<\/jats:italic>\n                      .\n                      <jats:italic>In vitro<\/jats:italic>\n                      validation confirmed dose-dependent effects: low-dose BMP4 (10\u00a0ng\/mL) sustained multipotency by upregulating both\n                      <jats:italic>RUNX2<\/jats:italic>\n                      and\n                      <jats:italic>SCX<\/jats:italic>\n                      , while high-dose BMP4 (100\u00a0ng\/mL) promoted tenogenic differentiation and suppressed osteogenic commitment. BMP4 overexpression induced nuclear SCX accumulation, confirming its pivotal role in tenogenic lineage commitment. Mechanistically, osteogenic differentiation required both BMPR1A and BMPR1B receptors, whereas tenogenic differentiation depended exclusively on BMPR1B signaling.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>These findings demonstrate that precise BMP4 dosage regulation dictates PDLSC differentiation outcomes, providing crucial insights for optimizing PDLSC-based periodontal regenerative therapies.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fbioe.2025.1738051","type":"journal-article","created":{"date-parts":[[2026,1,6]],"date-time":"2026-01-06T06:33:45Z","timestamp":1767681225000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["BMP4 dose dictates lineage specification bias in human periodontal ligament stem cells"],"prefix":"10.3389","volume":"13","author":[{"given":"Yihong","family":"Li","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qinghuang","family":"Tang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liwen","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chensheng","family":"Lin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Minhui","family":"Guo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanding","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fuhua","family":"Yan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Minkui","family":"Lin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,1,6]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1007\/s10266-015-0198-1","article-title":"Isolation, characterization and investigation of differentiation potential of human periodontal ligament cells and dental follicle progenitor cells and their response to BMP-7 in vitro","volume":"104","author":"Acil","year":"2016","journal-title":"Odontology"},{"key":"B2","doi-asserted-by":"publisher","first-page":"S39","DOI":"10.4103\/0975-7406.191964","article-title":"Bone morphogenetic proteins: signaling periodontal bone regeneration and repair","volume":"8","author":"Anusuya","year":"2016","journal-title":"J. Pharm. Bioallied Sci."},{"key":"B3","doi-asserted-by":"publisher","first-page":"32","DOI":"10.4236\/jbise.2013.68A1004","article-title":"BMP signaling in mesenchymal stem cell differentiation and bone formation","volume":"6","author":"Beederman","year":"2013","journal-title":"J. Biomed. Sci. Eng."},{"key":"B4","doi-asserted-by":"publisher","first-page":"e54115","DOI":"10.7759\/cureus.54115","article-title":"Advancements in periodontal regeneration: a comprehensive review of stem cell therapy","volume":"16","author":"Bharuka","year":"2024","journal-title":"Cureus"},{"key":"B5","volume-title":"EnhancedVolcano: publication-ready volcano plots with enhanced colouring and labeling","author":"Blighe","year":"2018"},{"key":"B6","doi-asserted-by":"publisher","first-page":"2680","DOI":"10.1242\/dev.093906","article-title":"Tendon-bone attachment unit is formed modularly by a distinct pool of Scx- and Sox9-positive progenitors","volume":"140","author":"Blitz","year":"2013","journal-title":"Development"},{"key":"B7","doi-asserted-by":"publisher","first-page":"963","DOI":"10.1016\/j.bioactmat.2020.06.023","article-title":"Advanced fabrication for electrospun three-dimensional nanofiber aerogels and scaffolds","volume":"5","author":"Chen","year":"2020","journal-title":"Bioact. Mater"},{"key":"B8","doi-asserted-by":"publisher","first-page":"165","DOI":"10.1186\/s13287-021-02227-7","article-title":"Effect of fibronectin, FGF-2, and BMP4 in the stemness maintenance of BMSCs and the metabolic and proteomic cues involved","volume":"12","author":"Chen","year":"2021","journal-title":"Stem Cell Res. Ther."},{"key":"B9","doi-asserted-by":"publisher","first-page":"1074","DOI":"10.7150\/thno.65694","article-title":"Single-cell RNA landscape of the osteoimmunology microenvironment in periodontitis","volume":"12","author":"Chen","year":"2022","journal-title":"Theranostics"},{"key":"B10","doi-asserted-by":"publisher","first-page":"102076","DOI":"10.1016\/j.jbc.2022.102076","article-title":"Type II BMP and activin receptors BMPR2 and ACVR2A share a conserved mode of growth factor recognition","volume":"298","author":"Chu","year":"2022","journal-title":"J. Biol. Chem."},{"key":"B11","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1016\/s1359-6101(99)00027-1","article-title":"The smads: transcriptional regulation and mouse models","volume":"11","author":"Datto","year":"2000","journal-title":"Cytokine Growth Factor Rev."},{"key":"B12","doi-asserted-by":"publisher","first-page":"4270","DOI":"10.1021\/acsabm.4c00613","article-title":"Nanofibrous microspheres: a biomimetic platform for bone tissue regeneration","volume":"7","author":"Desai","year":"2024","journal-title":"ACS Appl. Bio Mater"},{"key":"B13","doi-asserted-by":"publisher","first-page":"659","DOI":"10.1146\/annurev.cellbio.21.022404.142018","article-title":"Specificity and versatility in tgf-beta signaling through smads","volume":"21","author":"Feng","year":"2005","journal-title":"Annu. Rev. Cell Dev. Biol."},{"key":"B14","doi-asserted-by":"publisher","first-page":"149","DOI":"10.1111\/j.1601-6343.2007.00399.x","article-title":"Isolation and characterization of multipotent human periodontal ligament stem cells","volume":"10","author":"Gay","year":"2007","journal-title":"Orthod. Craniofac Res."},{"key":"B15","doi-asserted-by":"publisher","first-page":"119","DOI":"10.1002\/jbm.b.32988","article-title":"Bone morphogenetic protein-2, -6, and -7 differently regulate osteogenic differentiation of human periodontal ligament stem cells","volume":"102","author":"Hakki","year":"2014","journal-title":"J. Biomed. Mater Res. B Appl. Biomater."},{"key":"B16","doi-asserted-by":"publisher","first-page":"293","DOI":"10.1038\/s41587-023-01767-y","article-title":"Dictionary learning for integrative, multimodal and scalable single-cell analysis","volume":"42","author":"Hao","year":"2024","journal-title":"Nat. Biotechnol."},{"key":"B17","doi-asserted-by":"publisher","first-page":"1345","DOI":"10.1243\/09544119JEIM820","article-title":"Periodontal regeneration: a challenge for the tissue engineer?","volume":"224","author":"Hughes","year":"2010","journal-title":"Proc. Inst. Mech. Eng. H."},{"key":"B18","doi-asserted-by":"publisher","first-page":"550","DOI":"10.14348\/molcells.2017.0019","article-title":"Effect of FGF-2, TGF-beta-1, and BMPs on teno\/ligamentogenesis and osteo\/cementogenesis of human periodontal ligament stem cells","volume":"40","author":"Hyun","year":"2017","journal-title":"Mol. Cells"},{"key":"B19","doi-asserted-by":"publisher","first-page":"583","DOI":"10.4103\/0972-124X.119275","article-title":"Bone morphogenetic proteins: the anomalous molecules","volume":"17","author":"Jain","year":"2013","journal-title":"J. Indian Soc. Periodontol."},{"key":"B20","doi-asserted-by":"publisher","first-page":"1088","DOI":"10.1038\/s41467-021-21246-9","article-title":"Inference and analysis of cell-cell communication using CellChat","volume":"12","author":"Jin","year":"2021","journal-title":"Nat. Commun."},{"key":"B21","doi-asserted-by":"publisher","first-page":"337","DOI":"10.1002\/stem.2750","article-title":"Differences in the activity of endogenous bone morphogenetic protein signaling impact on the ability of induced pluripotent stem cells to differentiate to corneal epithelial-like cells","volume":"36","author":"Kamarudin","year":"2018","journal-title":"Stem Cells"},{"key":"B22","doi-asserted-by":"publisher","first-page":"43","DOI":"10.1007\/978-1-4419-1050-9_5","article-title":"Regulation of osteoblast differentiation by Runx2","volume":"658","author":"Komori","year":"2010","journal-title":"Adv. Exp. Med. Biol."},{"key":"B23","doi-asserted-by":"publisher","first-page":"e66107","DOI":"10.1371\/journal.pone.0066107","article-title":"Augmented BMPRIA-mediated BMP signaling in cranial neural crest lineage leads to cleft palate formation and delayed tooth differentiation","volume":"8","author":"Li","year":"2013","journal-title":"PLoS One"},{"key":"B24","doi-asserted-by":"publisher","first-page":"1670","DOI":"10.1089\/scd.2012.0548","article-title":"Effect of bone morphogenetic protein-4 on the expression of Sox2, Oct-4, and c-Myc in human periodontal ligament cells during long-term culture","volume":"22","author":"Liu","year":"2013","journal-title":"Stem Cells Dev."},{"key":"B25","doi-asserted-by":"publisher","first-page":"550","DOI":"10.1186\/s13059-014-0550-8","article-title":"Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2","volume":"15","author":"Love","year":"2014","journal-title":"Genome Biol."},{"key":"B26","doi-asserted-by":"publisher","first-page":"2783","DOI":"10.1101\/gad.1350705","article-title":"Smad transcription factors","volume":"19","author":"Massague","year":"2005","journal-title":"Genes Dev."},{"key":"B27","doi-asserted-by":"publisher","first-page":"1017613","DOI":"10.3389\/fbioe.2022.1017613","article-title":"Stem cell homing in periodontal tissue regeneration","volume":"10","author":"Meng","year":"2022","journal-title":"Front. Bioeng. Biotechnol."},{"key":"B28","doi-asserted-by":"publisher","first-page":"1001","DOI":"10.1089\/scd.2013.0490","article-title":"Periodontal-ligament-derived stem cells exhibit the capacity for long-term survival, self-renewal, and regeneration of multiple tissue types in vivo","volume":"23","author":"Menicanin","year":"2014","journal-title":"Stem Cells Dev."},{"key":"B29","doi-asserted-by":"publisher","first-page":"e10172","DOI":"10.1002\/jbm4.10172","article-title":"Heterotopic ossification: a comprehensive review","volume":"3","author":"Meyers","year":"2019","journal-title":"JBMR Plus"},{"key":"B30","doi-asserted-by":"publisher","first-page":"333","DOI":"10.1016\/j.reth.2025.01.002","article-title":"The next generation of regenerative dentistry: from tooth development biology to periodontal tissue, dental pulp, and whole tooth reconstruction in the clinical setting","volume":"28","author":"Morita","year":"2025","journal-title":"Regen. Ther."},{"key":"B31","doi-asserted-by":"publisher","first-page":"72","DOI":"10.1016\/j.reth.2019.12.011","article-title":"Mesenchymal stem cell-based tissue regeneration therapies for periodontitis","volume":"14","author":"Ouchi","year":"2020","journal-title":"Regen. Ther."},{"key":"B32","doi-asserted-by":"publisher","first-page":"1374","DOI":"10.1007\/s10439-023-03173-6","article-title":"Role and mechanism of BMP4 in regenerative medicine and tissue engineering","volume":"51","author":"Pan","year":"2023","journal-title":"Ann. Biomed. Eng."},{"key":"B33","doi-asserted-by":"publisher","first-page":"121","DOI":"10.5051\/jpis.2020.50.2.121","article-title":"Enhancement of peri-implant bone formation via parathyroid hormone administration in a rat model at risk for medication-related osteonecrosis of the jaw","volume":"50","author":"Park","year":"2020","journal-title":"J. Periodontal Implant Sci."},{"key":"B34","doi-asserted-by":"publisher","first-page":"399","DOI":"10.1038\/s41580-018-0002-5","article-title":"Single-cell transcriptional profiling: a window into embryonic cell-type specification","volume":"19","author":"Pijuan-Sala","year":"2018","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"B35","doi-asserted-by":"publisher","first-page":"979","DOI":"10.1038\/nmeth.4402","article-title":"Reversed graph embedding resolves complex single-cell trajectories","volume":"14","author":"Qiu","year":"2017","journal-title":"Nat. Methods"},{"key":"B36","doi-asserted-by":"publisher","first-page":"e1062","DOI":"10.4317\/medoral.17925","article-title":"Mesenchymal dental stem cells in regenerative dentistry","volume":"17","author":"Rodriguez-Lozano","year":"2012","journal-title":"Med. Oral Patol. Oral Cir. Bucal"},{"key":"B37","doi-asserted-by":"publisher","first-page":"2","DOI":"10.3390\/jfb16010002","article-title":"Nano delivery system for atherosclerosis","volume":"16","author":"Rong","year":"2024","journal-title":"J. Funct. Biomater."},{"key":"B38","doi-asserted-by":"publisher","first-page":"20220706","DOI":"10.1515\/biol-2022-0706","article-title":"Signalling pathways in the osteogenic differentiation of periodontal ligament stem cells","volume":"18","author":"Ru","year":"2023","journal-title":"Open Life Sci."},{"key":"B39","doi-asserted-by":"publisher","first-page":"17","DOI":"10.1016\/j.diff.2022.02.003","article-title":"BMP-2 and asporin expression regulate 5-aza-dC-mediated osteoblast\/cementoblast differentiation of periodontal dental ligament mesenchymal progenitor cells","volume":"124","author":"Sacramento","year":"2022","journal-title":"Differentiation"},{"key":"B40","doi-asserted-by":"publisher","first-page":"3855","DOI":"10.1242\/dev.128.19.3855","article-title":"Analysis of the tendon cell fate using scleraxis, a specific marker for tendons and ligaments","volume":"128","author":"Schweitzer","year":"2001","journal-title":"Development"},{"key":"B41","doi-asserted-by":"publisher","first-page":"e24712","DOI":"10.7554\/eLife.24712","article-title":"Resolving stem and progenitor cells in the adult mouse incisor through gene co-expression analysis","volume":"6","author":"Seidel","year":"2017","journal-title":"Elife"},{"key":"B42","doi-asserted-by":"publisher","first-page":"149","DOI":"10.1016\/S0140-6736(04)16627-0","article-title":"Investigation of multipotent postnatal stem cells from human periodontal ligament","volume":"364","author":"Seo","year":"2004","journal-title":"Lancet"},{"key":"B43","doi-asserted-by":"publisher","first-page":"1045103","DOI":"10.3389\/fcell.2022.1045103","article-title":"The role of BMP4 in adipose-derived stem cell differentiation: a minireview","volume":"10","author":"Setiawan","year":"2022","journal-title":"Front. Cell Dev. Biol."},{"key":"B44","doi-asserted-by":"publisher","first-page":"433","DOI":"10.1186\/s12859-021-04344-9","article-title":"CellProfiler 4: improvements in speed, utility and usability","volume":"22","author":"Stirling","year":"2021","journal-title":"BMC Bioinforma."},{"key":"B45","doi-asserted-by":"publisher","first-page":"44","DOI":"10.1038\/s41368-025-00371-x","article-title":"The research progress on periodontitis by the National Natural Science Foundation of China","volume":"17","author":"Xie","year":"2025","journal-title":"Int. J. Oral Sci."},{"key":"B46","doi-asserted-by":"publisher","first-page":"71","DOI":"10.1177\/00220345231205283","article-title":"Single-cell transcriptomic analysis of dental pulp and periodontal ligament stem cells","volume":"103","author":"Yang","year":"2024","journal-title":"J. Dent. Res."},{"key":"B47","doi-asserted-by":"publisher","first-page":"1065","DOI":"10.7150\/ijms.8473","article-title":"Bone morphogenetic protein-9 induces PDLSCs osteogenic differentiation through the ERK and p38 signal pathways","volume":"11","author":"Ye","year":"2014","journal-title":"Int. J. Med. Sci."},{"key":"B48","doi-asserted-by":"publisher","first-page":"284","DOI":"10.1089\/omi.2011.0118","article-title":"clusterProfiler: an R package for comparing biological themes among gene clusters","volume":"16","author":"Yu","year":"2012","journal-title":"OMICS"},{"key":"B49","doi-asserted-by":"publisher","first-page":"150","DOI":"10.1002\/jcp.24693","article-title":"Effect of CTGF\/CCN2 on osteo\/cementoblastic and fibroblastic differentiation of a human periodontal ligament stem\/progenitor cell line","volume":"230","author":"Yuda","year":"2015","journal-title":"J. Cell Physiol."},{"key":"B50","doi-asserted-by":"publisher","first-page":"93","DOI":"10.1016\/j.archoralbio.2019.04.001","article-title":"MicroRNA-23a inhibits osteogenesis of periodontal mesenchymal stem cells by targeting bone morphogenetic protein signaling","volume":"102","author":"Zhang","year":"2019","journal-title":"Arch. Oral Biol."},{"key":"B51","doi-asserted-by":"publisher","first-page":"108069","DOI":"10.1016\/j.celrep.2020.108069","article-title":"Unsupervised inference of developmental directions for single cells using VECTOR","volume":"32","author":"Zhang","year":"2020","journal-title":"Cell Rep."},{"key":"B52","doi-asserted-by":"publisher","first-page":"653","DOI":"10.1186\/s13287-025-04777-6","article-title":"Periodontal ligament stem cells in tissue remodeling: from mechanical forces to inflammatory signals","volume":"16","author":"Zhao","year":"2025","journal-title":"Stem Cell Res. Ther."},{"key":"B53","doi-asserted-by":"publisher","first-page":"972313","DOI":"10.1155\/2015\/972313","article-title":"Periodontal ligament stem cells: current status, concerns, and future prospects","volume":"2015","author":"Zhu","year":"2015","journal-title":"Stem Cells Int."}],"container-title":["Frontiers in Bioengineering and Biotechnology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fbioe.2025.1738051\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,6]],"date-time":"2026-01-06T06:33:48Z","timestamp":1767681228000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fbioe.2025.1738051\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,6]]},"references-count":53,"alternative-id":["10.3389\/fbioe.2025.1738051"],"URL":"https:\/\/doi.org\/10.3389\/fbioe.2025.1738051","relation":{},"ISSN":["2296-4185"],"issn-type":[{"value":"2296-4185","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,6]]},"article-number":"1738051"}}