{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T20:54:56Z","timestamp":1790628896958,"version":"4.1.0"},"reference-count":105,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T00:00:00Z","timestamp":1782259200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T00:00:00Z","timestamp":1782259200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100003803","name":"University of Hong Kong","doi-asserted-by":"publisher","award":["2302101543"],"award-info":[{"award-number":["2302101543"]}],"id":[{"id":"10.13039\/501100003803","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Int Endodontic J"],"published-print":{"date-parts":[[2026,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Dental pulp inflammation triggers immune responses involving macrophages and dental pulp stem cells (DPSCs), which interact to regulate angiogenesis essential for tissue repair. M1 pro\u2010inflammatory macrophages predominate early in pulpitis, and clarifying their angiogenic role is vital in identifying inflammatory regenerative mechanisms.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methodology<\/jats:title>\n                    <jats:p>\n                      THP\u20101 cells and peripheral blood monocyte (PBM)\u2010derived macrophages were polarized to M1 or M2 phenotypes, characterized by qRT\u2010PCR, ELISA, and angiogenesis arrays. A vasculature\u2010on\u2010a\u2010chip comprising DPSCs, human umbilical vein endothelial cells (HUVECs) and THP\u20101\u2010derived macrophages was imaged, and the vascular segments\/sprouts were quantified using ImageJ. Density effects used 5\u2009\u00d7\u200910\n                      <jats:sup>4<\/jats:sup>\n                      versus 7.5\u2009\u00d7\u200910\n                      <jats:sup>4<\/jats:sup>\n                      \u2009M1 macrophages\/device, with propidium iodide staining for cytotoxicity. IL\u20108 effects on DPSC VEGF secretion were assessed by ELISA (with\/without Reparaxin 1\u2009\u03bcM), Matrigel tube formation assays, and exogenous IL\u20108 (0.5\u2009ng\/mL). Transwell co\u2010cultures underwent RNA sequencing and bioinformatics analysis, which identified candidate hub genes and signalling pathways; the results were validated by Western blotting (p\u2010ERK, HIF\u20101\u03b1; ERK inhibitor SCH772984, 25\u2009nM). Statistical testing was performed using ANOVA with Tukey's post hoc test (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.05).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      M1 macrophages at low density (5\u2009\u00d7\u200910\n                      <jats:sup>4<\/jats:sup>\n                      cells\/device) significantly enhanced vascularization in the vasculature\u2010on\u2010a\u2010chip, increasing vascular segments (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.0001) and free sprouts (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.05\u20130.01) compared to M0 or no\u2010macrophage controls, with effects comparable to M2. High\u2010density M1 seeding (7.5\u2009\u00d7\u200910\n                      <jats:sup>4<\/jats:sup>\n                      cells\/device) reduced sprouts (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.0001\u2009day 4,\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.01\u2009day 5) due to increased cytotoxicity (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.0001). Both THP\u20101\u2010 and PBM\u2010derived M1\u2010conditioned media (CM) showed significantly elevated IL\u20108 levels. M1 CM (THP\u20101\/PBM) induced DPSC VEGF secretion, blocked by Reparaxin (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.01\u20130.0001), confirming IL\u20108 mediation via CXCR1\/2. Exogenous IL\u20108 (0.5\u2009ng\/mL) upregulated DPSC VEGF protein\/mRNA (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.05) and Matrigel tube formation (segments\/junctions\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.05). M1\u2010DPSC CM enhanced vascular meshes\/segments on Matrigel (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.05), reduced by Reparaxin. RNA\u2010seq of M1 co\u2010cultured DPSCs identified 17 angiogenic genes (logFC &gt;\u20091.2), with HIF\u20101\u03b1 as a hub gene and an enriched MAPK\/ERK pathway. Western blot analysis confirmed MAPK\/ERK\u2010HIF\u20101\u03b1 as a contributory pathway in IL\u20108\u2010induced upregulation of VEGF in DPSCs.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>M1 macrophages promote angiogenesis via IL\u20108\u2010induced DPSC VEGF secretion through CXCR1\/2\u2010MAPK\/ERK\u2010HIF\u20101\u03b1 signalling density\u2010dependently, suggesting that therapeutic modulation rather than total suppression of M1 activity could improve outcomes in vital pulp therapy.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1111\/iej.70198","type":"journal-article","created":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T09:18:11Z","timestamp":1782292691000},"page":"2143-2163","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["The Interplay of\n                    <scp>M1<\/scp>\n                    Macrophages and Dental Pulp Stem Cells Promotes Angiogenesis Through\n                    <scp>IL<\/scp>\n                    \u20108\u2010Dependent\n                    <scp>VEGF<\/scp>\n                    Regulation: An In\u00a0Vitro Study"],"prefix":"10.1111","volume":"59","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5420-9879","authenticated-orcid":false,"given":"Dineshi Sewvandi","family":"Thalakiriyawa","sequence":"first","affiliation":[{"name":"Applied Oral Sciences &amp; Community Dental Care, Faculty of Dentistry The University of Hong Kong  Hong Kong SAR China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohamad","family":"Koohi\u2010Moghadam","sequence":"additional","affiliation":[{"name":"Department of Diagnostic Radiology, Li K Shing Faculty of Medicine The University of Hong Kong  Hong Kong SAR China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingxin","family":"Hu","sequence":"additional","affiliation":[{"name":"Restorative Dental Sciences, Faculty of Dentistry The University of Hong Kong  Hong Kong SAR China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hong","family":"Wang","sequence":"additional","affiliation":[{"name":"Applied Oral Sciences &amp; Community Dental Care, Faculty of Dentistry The University of Hong Kong  Hong Kong SAR China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9379-0889","authenticated-orcid":false,"given":"Colman","family":"McGrath","sequence":"additional","affiliation":[{"name":"Applied Oral Sciences &amp; Community Dental Care, Faculty of Dentistry The University of Hong Kong  Hong Kong SAR China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3621-4866","authenticated-orcid":false,"given":"Waruna Lakmal","family":"Dissanayaka","sequence":"additional","affiliation":[{"name":"Applied Oral Sciences &amp; Community Dental Care, Faculty of Dentistry The University of Hong Kong  Hong Kong SAR 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