{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:54:02Z","timestamp":1785405242562,"version":"3.56.0"},"reference-count":28,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T00:00:00Z","timestamp":1777420800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006345","name":"Nicolaus Copernicus University","doi-asserted-by":"publisher","award":["51\/2024\/Grants4NCUStudents"],"award-info":[{"award-number":["51\/2024\/Grants4NCUStudents"]}],"id":[{"id":"10.13039\/501100006345","id-type":"DOI","asserted-by":"publisher"}]},{"award":["51\/2024\/Grants4NCUStudents"],"award-info":[{"award-number":["51\/2024\/Grants4NCUStudents"]}],"id":[{"id":"https:\/\/ror.org\/0102mm775","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Transforming growth factor-\u03b21 (TGF-\u03b21) and tumor necrosis factor-\u03b1 (TNF-\u03b1) are central regulators of vascular inflammation and remodeling in coronary artery disease. However, their cell-type-specific and context-dependent effects in primary human coronary artery endothelial cells (ECs) and vascular smooth muscle cells (VSMCs) remain incompletely defined. Primary human coronary artery endothelial cells (pHCAECs) and smooth muscle cells (pHCASMCs) were stimulated with TGF-\u03b21 (10 ng\/mL), TNF-\u03b1 (100 ng\/mL), or their combination. Canonical SMAD2\/3 activation, Kr\u00fcppel-like factor 11 (KLF11) expression, cytoskeletal and junctional remodeling, vascular cell adhesion molecule-1 (VCAM-1) expression, migration dynamics (wound healing and confluent assays), and endothelial tube formation were assessed using immunofluorescence microscopy, live-cell imaging, and quantitative trajectory analysis. Both cytokines were associated with increased nuclear pSMAD2\/3 signal in ECs and VSMCs, consistent with functional interplay between inflammatory and TGF-\u03b2-related signaling pathways. In pHCAECs, TNF-\u03b1 robustly induced VCAM-1 functional expression and disrupted VE-cadherin continuity, whereas TGF-\u03b21 primarily promoted cytoskeletal remodeling without strong inflammatory activation. TGF-\u03b21 increased endothelial migration velocity and accumulated distance. In contrast, TNF-\u03b1 preferentially enhanced Euclidean displacement and directional persistence, shifting the migratory pattern toward more directed movement most evident under combined TGF-\u03b21 + TNF-\u03b1 stimulation. Notably, TGF-\u03b21 significantly reduced endothelial tube formation, indicating impaired network organization rather than proangiogenic activity. In pHCASMCs, TGF-\u03b21 enhanced migratory activity, particularly in confluent monolayers, whereas TNF-\u03b1 enhanced directional displacement. KLF11 was induced by TGF-\u03b21 in both pHCAECs and pHCASMCs. In pHCAECs, TNF-\u03b1 also increased KLF11 and co-stimulation promoted nuclear enrichment, whereas in pHCASMCs TNF-\u03b1 alone was not effective and combined treatment amplified the TGF-\u03b21 response, supporting cell-type-specific integration of inflammatory and TGF-\u03b2-dependent signals. TGF-\u03b21 and TNF-\u03b1 differentially regulate the inflammatory activation and migration of primary human coronary vascular cells in a cell-type- and structural-context-dependent manner. TGF-\u03b21 enhances migratory force generation, whereas TNF-\u03b1 reinforces directional polarization, and their integration determines effective vascular repair dynamics. Canonical SMAD2\/3 activation does not uniformly predict functional outcome, and KLF11 was identified as a context-sensitive transcription-associated factor showing differential nuclear localization in response to cytokine stimulation, representing a hypothesis-generating observation for future mechanistic studies.<\/jats:p>","DOI":"10.3390\/ijms27093948","type":"journal-article","created":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T09:46:11Z","timestamp":1777455971000},"page":"3948","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["TGF-\u03b2 and TNF-\u03b1 Signaling Crosstalk in Human Coronary Artery Cells"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2098-2799","authenticated-orcid":false,"given":"Klaudia","family":"Bonowicz-Koz\u0142owska","sequence":"first","affiliation":[{"name":"Department of Histology and Embryology, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, 85-092 Bydgoszcz, Poland"},{"name":"Department of Morphological and Physiological Sciences, Faculty of Medicine, Collegium Medicum, Mazovian Academy in P\u0142ock, 09-402 P\u0142ock, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-3777-598X","authenticated-orcid":false,"given":"Dominika","family":"Jerka","sequence":"additional","affiliation":[{"name":"Department of Histology and Embryology, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, 85-092 Bydgoszcz, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Damian","family":"Twardak","sequence":"additional","affiliation":[{"name":"Department of Histology and Embryology, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, 85-092 Bydgoszcz, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1311-263X","authenticated-orcid":false,"given":"Konrad","family":"Kleszczy\u0144ski","sequence":"additional","affiliation":[{"name":"Department of Dermatology, University of M\u00fcnster, Von-Esmarch-Str. 58, 48149 M\u00fcnster, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5445-8821","authenticated-orcid":false,"given":"Maciej","family":"Gagat","sequence":"additional","affiliation":[{"name":"Department of Histology and Embryology, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, 85-092 Bydgoszcz, Poland"},{"name":"Department of Morphological and Physiological Sciences, Faculty of Medicine, Collegium Medicum, Mazovian Academy in P\u0142ock, 09-402 P\u0142ock, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2383","DOI":"10.1016\/j.immuni.2025.09.012","article-title":"Inflammation in atherosclerosis: Lessons and therapeutic implications","volume":"58","author":"Libby","year":"2025","journal-title":"Immunity"},{"key":"ref_2","first-page":"2965","article-title":"Tropomyosin\u2014Master regulator of actin filament function in the cytoskeleton","volume":"128","author":"Gunning","year":"2015","journal-title":"J. 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