{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T13:37:12Z","timestamp":1785418632282,"version":"3.56.0"},"reference-count":36,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,4,11]],"date-time":"2026-04-11T00:00:00Z","timestamp":1775865600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"European Union, Next Generation EU","award":["PNRR-MAD-2022-12376878"],"award-info":[{"award-number":["PNRR-MAD-2022-12376878"]}]},{"award":["PNRR-MAD-2022-12376878"],"award-info":[{"award-number":["PNRR-MAD-2022-12376878"]}],"id":[{"id":"https:\/\/ror.org\/019w4f821","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Inflammation is associated with metabolic alterations that can lead to the release of volatile organic compounds (VOCs) reflecting cellular biochemical activity. Profiling these volatile metabolites may provide insight into cellular responses to inflammatory stimuli, although their characterization in skin-derived cells remains limited. In this exploratory proof-of-concept study, we investigated the volatile metabolite profiles of human skin fibroblasts exposed to different inflammatory stimuli. Fibroblast cell lines were stimulated with polyinosinic:polycytidylic acid (Poly I:C), tumor necrosis factor-alpha (TNF-\u03b1), and lipopolysaccharide (LPS) to model viral-, cytokine-, and bacterial-associated stress conditions. Headspace solid-phase microextraction coupled with comprehensive two-dimensional gas chromatography and time-of-flight mass spectrometry (HS-SPME-GC\u00d7GC-TOFMS) was applied to analyze volatile metabolites released from the cell cultures, enabling exploratory profiling of the fibroblast volatilome. A data-processing workflow including pairwise comparisons between experimental groups and statistical filtering was implemented to identify volatile features associated with the different conditions. Several VOCs were tentatively identified, mainly belonging to alcohol, ester, and hydrocarbon classes, and showed differential abundance patterns between stimulated and control samples. Multivariate analysis indicated a separation between stimulated and non-stimulated groups, suggesting stimulus-associated differences in the volatile profiles of fibroblast cultures. While these observations may reflect metabolic responses occurring under inflammatory stimulation, the chemical identity and biochemical origins of several detected features remain to be confirmed. All in all, this study demonstrates the feasibility of applying HS-SPME-GC\u00d7GC-TOFMS-based volatilome profiling to investigate stimulus-associated changes in fibroblast cultures. The detected VOC patterns should therefore be considered preliminary observations requiring further chemical characterization and independent validation. Future studies including larger sample numbers, complementary biological verification of the inflammatory response, and more physiologically relevant experimental models will be necessary to further assess the robustness and potential relevance of these volatile signatures in the context of inflammatory processes.<\/jats:p>","DOI":"10.3390\/ijms27083429","type":"journal-article","created":{"date-parts":[[2026,4,13]],"date-time":"2026-04-13T08:49:02Z","timestamp":1776070142000},"page":"3429","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Exploratory Volatilome Profiling of Inflammation in Skin Fibroblasts: A Proof-of-Concept Study"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-1379-6805","authenticated-orcid":false,"given":"Riccardo","family":"Di Stefano","sequence":"first","affiliation":[{"name":"Department of Chemical, Pharmaceutical and Agricultural Sciences, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-2582-6025","authenticated-orcid":false,"given":"Marco","family":"De Poli","sequence":"additional","affiliation":[{"name":"Department of Chemical, Pharmaceutical and Agricultural Sciences, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0060-5870","authenticated-orcid":false,"given":"Chiara","family":"Moltrasio","sequence":"additional","affiliation":[{"name":"Dermatology Unit, Fondazione IRCCS Ca\u2019 Granda Ospedale Maggiore Policlinico, 20122 Milan, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8160-4169","authenticated-orcid":false,"given":"Angelo V.","family":"Marzano","sequence":"additional","affiliation":[{"name":"Dermatology Unit, Fondazione IRCCS Ca\u2019 Granda Ospedale Maggiore Policlinico, 20122 Milan, Italy"},{"name":"Department of Pathophysiology and Transplantation, Universit\u00e0 Degli Studi di Milano, 20122 Milan, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8241-2513","authenticated-orcid":false,"given":"Erika","family":"Rimondi","sequence":"additional","affiliation":[{"name":"Department of Translational Medicine and LTTA Centre, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7829-0824","authenticated-orcid":false,"given":"Elisabetta","family":"Melloni","sequence":"additional","affiliation":[{"name":"Department of Translational Medicine and LTTA Centre, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paola","family":"Secchiero","sequence":"additional","affiliation":[{"name":"Department of Translational Medicine and LTTA Centre, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-8491-6035","authenticated-orcid":false,"given":"Giada","family":"Lodi","sequence":"additional","affiliation":[{"name":"Department of Translational Medicine and LTTA Centre, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-4216-4489","authenticated-orcid":false,"given":"Marta","family":"Manfredini","sequence":"additional","affiliation":[{"name":"Department of Translational Medicine, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alberto","family":"Cavazzini","sequence":"additional","affiliation":[{"name":"Department of Chemical, Pharmaceutical and Agricultural Sciences, University of Ferrara, 44121 Ferrara, Italy"},{"name":"Council for Agricultural Research and Economics (CREA), 00184 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3427-618X","authenticated-orcid":false,"given":"Annalisa","family":"Marcuzzi","sequence":"additional","affiliation":[{"name":"Department of Translational Medicine, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8493-1168","authenticated-orcid":false,"given":"Sergio","family":"Crovella","sequence":"additional","affiliation":[{"name":"Department of Environmental and Prevention Sciences, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7236-4266","authenticated-orcid":false,"given":"Flavio A.","family":"Franchina","sequence":"additional","affiliation":[{"name":"Department of Chemical, Pharmaceutical and Agricultural Sciences, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"802","DOI":"10.1016\/j.jid.2017.09.045","article-title":"SFRP2\/DPP4 and FMO1\/LSP1 Define Major Fibroblast Populations in Human Skin","volume":"138","author":"Tabib","year":"2018","journal-title":"J. Investig. Dermatol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1735","DOI":"10.1016\/j.jid.2020.11.028","article-title":"Human Dermal Fibroblast Subpopulations Are Conserved across Single-Cell RNA Sequencing Studies","volume":"141","author":"Izeta","year":"2021","journal-title":"J. Investig. Dermatol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"eaba6500","DOI":"10.1126\/science.aba6500","article-title":"Developmental cell programs are co-opted in inflammatory skin disease","volume":"371","author":"Reynolds","year":"2021","journal-title":"Science"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1807","DOI":"10.1038\/s41590-025-02267-8","article-title":"A single-cell and spatial genomics atlas of human skin fibroblasts reveals shared disease-related fibroblast subtypes across tissues","volume":"26","author":"Steele","year":"2025","journal-title":"Nat. Immunol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"19385","DOI":"10.1038\/s41598-021-98651-z","article-title":"TNF induces glycolytic shift in fibroblast like synoviocytes via GLUT1 and HIF1A","volume":"11","author":"Koedderitzsch","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"5555","DOI":"10.1111\/febs.15644","article-title":"Metabolic dysfunction and inflammatory disease: The role of stromal fibroblasts","volume":"288","author":"Farah","year":"2021","journal-title":"FEBS J."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"00427","DOI":"10.1183\/23120541.00427-2022","article-title":"Validation of volatile metabolites of pulmonary oxidative injury: A bench to bedside study","volume":"9","author":"Fenn","year":"2023","journal-title":"ERJ Open Res."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"M\u00fcller-Wirtz, L.M., Kiefer, D., Ruffing, S., Brausch, T., H\u00fcppe, T., Sessler, D.I., Volk, T., Fink, T., Kreuer, S., and Maurer, F. (2021). Quantification of Volatile Aldehydes Deriving from In Vitro Lipid Peroxidation in the Breath of Ventilated Patients. Molecules, 26.","DOI":"10.3390\/molecules26113089"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Kononova, E., Me\u017emale, L., Po\u013caka, I., Veliks, V., Anarkulova, L., Vilkoite, I., Tolmanis, I., \u013be\u0161\u010dinska, A.M., Ston\u0101ns, I., and P\u010dolkins, A. (2024). Breath Fingerprint of Colorectal Cancer Patients Based on the Gas Chromatography\u2013Mass Spectrometry Analysis. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25031632"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Albrecht, F.W., Maurer, F., M\u00fcller-Wirtz, L.M., Schwaiblmair, M.H., H\u00fcppe, T., Wolf, B., Sessler, D.I., Volk, T., Kreuer, S., and Fink, T. (2020). Exhaled Volatile Organic Compounds during Inflammation Induced by TNF-\u03b1 in Ventilated Rats. Metabolites, 10.","DOI":"10.3390\/metabo10060245"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1007\/s11306-024-02180-5","article-title":"The human volatilome meets cancer diagnostics: Past, present, and future of noninvasive applications","volume":"20","author":"Barbosa","year":"2024","journal-title":"Metabolomics"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Tsou, P.-H., Lin, Z.-L., Pan, Y.-C., Yang, H.-C., Chang, C.-J., Liang, S.-K., Wen, Y.-F., Chang, C.-H., Chang, L.-Y., and Yu, K.-L. (2021). Exploring Volatile Organic Compounds in Breath for High-Accuracy Prediction of Lung Cancer. Cancers, 13.","DOI":"10.3390\/cancers13061431"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"334","DOI":"10.1111\/apt.15622","article-title":"Volatile organic compounds in breath can serve as a non-invasive diagnostic biomarker for the detection of advanced adenomas and colorectal cancer","volume":"51","author":"Jansen","year":"2020","journal-title":"Aliment. Pharmacol. Ther."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"5949","DOI":"10.1021\/cr300174a","article-title":"Volatile Organic Compounds of Lung Cancer and Possible Biochemical Pathways","volume":"112","author":"Hakim","year":"2012","journal-title":"Chem. Rev."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1007\/s11306-024-02142-x","article-title":"Progress and challenges of developing volatile metabolites from exhaled breath as a biomarker platform","volume":"20","author":"Chou","year":"2024","journal-title":"Metabolomics"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1159\/000522417","article-title":"Metabolic Regulation of Fibroblast Activation and Proliferation during Organ Fibrosis","volume":"8","author":"Wang","year":"2022","journal-title":"Kidney Dis."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"de Jong, F.J.M., Lilien, T.A., Fenn, D.W., Wingelaar, T.T., van Ooij, P.-J.A.M., der Zee, A.H.M.-V., Hollmann, M.W., van Hulst, R.A., and Brinkman, P. (2024). Volatile Organic Compounds in Cellular Headspace after Hyperbaric Oxygen Exposure: An In Vitro Pilot Study. Metabolites, 14.","DOI":"10.3390\/metabo14050281"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"732","DOI":"10.1038\/35099560","article-title":"Recognition of double-stranded RNA and activation of NF-\u03baB by Toll-like receptor 3","volume":"413","author":"Alexopoulou","year":"2001","journal-title":"Nature"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1038\/nrrheum.2015.169","article-title":"TNF biology, pathogenic mechanisms and emerging therapeutic strategies","volume":"12","author":"Kalliolias","year":"2016","journal-title":"Nat. Rev. Rheumatol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1111\/wrr.12985","article-title":"Role of haematopoietic cell-specific protein 1-associated protein X-1 gene in lipopolysaccharide-induced apoptosis of human dermal fibroblasts","volume":"30","author":"Ning","year":"2022","journal-title":"Wound Repair. Regen."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1016\/j.jchromb.2009.12.033","article-title":"Volatile profiles of human skin cell cultures in different degrees of senescence","volume":"878","author":"Acevedo","year":"2010","journal-title":"J. Chromatogr. B."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1016\/j.trac.2018.10.015","article-title":"Advances in the application of comprehensive two-dimensional gas chromatography in metabolomics","volume":"109","author":"Keppler","year":"2018","journal-title":"TrAC Trends Anal. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Szymczak, K., Nawrocka, J., and Bonikowski, R. (2024). Fingerprinting of Volatile Organic Compounds in Old and Commercial Apple Cultivars by HS-SPME GC\/GC-ToF-MS. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms252413478"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1002\/ansa.202000142","article-title":"30th Anniversary of comprehensive two-dimensional gas chromatography: Latest advances","volume":"2","author":"Zanella","year":"2021","journal-title":"Anal. Sci. Adv."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Shi, Z., Liu, Z., Wei, Y., Zhang, R., Deng, Y., and Li, D. (2024). The role of dermal fibroblasts in autoimmune skin diseases. Front. Immunol., 15.","DOI":"10.3389\/fimmu.2024.1379490"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"274","DOI":"10.1007\/s12016-024-08997-1","article-title":"Fibroblast: A Novel Target for Autoimmune and Inflammatory Skin Diseases Therapeutics","volume":"66","author":"Chen","year":"2024","journal-title":"Clin. Rev. Allergy Immunol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1007","DOI":"10.1081\/SAC-200040345","article-title":"Some Modifications on the Application of the Exact Wilcoxon\u2013Mann\u2013Whitney Test","volume":"33","author":"Chen","year":"2004","journal-title":"Commun. Stat. Simul. Comput."},{"key":"ref_28","first-page":"72","article-title":"Different Outcomes of the Wilcoxon\u2014Mann\u2014Whitney Test from Different Statistics Packages","volume":"54","author":"Bergmann","year":"2000","journal-title":"Am. Stat."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Abaffy, T., Duncan, R., Riemer, D.D., Tietje, O., Elgart, G., Milikowski, C., and DeFazio, R.A. (2010). Differential Volatile Signatures from Skin, Naevi and Melanoma: A Novel Approach to Detect a Pathological Process. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0013813"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"7238","DOI":"10.1038\/s41598-024-57967-2","article-title":"Effects of psychological stress on the emission of volatile organic compounds from the skin","volume":"14","author":"Lucchi","year":"2024","journal-title":"Sci. Rep."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1016\/j.biochi.2023.09.004","article-title":"Ether-lipids and cellular signaling: A differential role of alkyl- and alkenyl-ether-lipids?","volume":"215","author":"Papin","year":"2023","journal-title":"Biochimie"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1016\/j.aca.2018.03.037","article-title":"SPME-GC\u00d7GC-TOF MS fingerprint of virally-infected cell culture: Sample preparation optimization and data processing evaluation","volume":"1027","author":"Purcaro","year":"2018","journal-title":"Anal. Chim. Acta"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1016\/j.foodchem.2016.07.088","article-title":"Coffee aroma: Chemometric comparison of the chemical information provided by three different samplings combined with GC\u2013MS to describe the sensory properties in cup","volume":"214","author":"Bressanello","year":"2017","journal-title":"Food Chem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.chroma.2004.12.084","article-title":"Equilibrium in-fibre standardisation technique for solid-phase microextraction","volume":"1072","author":"Wang","year":"2005","journal-title":"J. Chromatogr. A"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1097\/00001648-199001000-00010","article-title":"No adjustments are needed for multiple comparisons","volume":"1","author":"Rothman","year":"1990","journal-title":"Epidemiology"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1016\/S0895-4356(00)00314-0","article-title":"Adjusting for multiple testing\u2014When and how?","volume":"54","author":"Bender","year":"2001","journal-title":"J. Clin. Epidemiol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/8\/3429\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,13]],"date-time":"2026-04-13T08:52:16Z","timestamp":1776070336000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/8\/3429"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,11]]},"references-count":36,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,4]]}},"alternative-id":["ijms27083429"],"URL":"https:\/\/doi.org\/10.3390\/ijms27083429","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,11]]}}}