{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,8]],"date-time":"2026-06-08T13:41:28Z","timestamp":1780926088786,"version":"3.54.1"},"reference-count":47,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2023,10,16]],"date-time":"2023-10-16T00:00:00Z","timestamp":1697414400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100009566","name":"UiO: Life Science at the University of Oslo","doi-asserted-by":"publisher","award":["2018\/10221"],"award-info":[{"award-number":["2018\/10221"]}],"id":[{"id":"10.13039\/501100009566","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100009566","name":"UiO: Life Science at the University of Oslo","doi-asserted-by":"publisher","award":["2019050"],"award-info":[{"award-number":["2019050"]}],"id":[{"id":"10.13039\/501100009566","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006095","name":"South-Eastern Norway Regional Health Authority","doi-asserted-by":"publisher","award":["2018\/10221"],"award-info":[{"award-number":["2018\/10221"]}],"id":[{"id":"10.13039\/501100006095","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006095","name":"South-Eastern Norway Regional Health Authority","doi-asserted-by":"publisher","award":["2019050"],"award-info":[{"award-number":["2019050"]}],"id":[{"id":"10.13039\/501100006095","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Cytokines are mediators of inflammation that could lead to fibrosis. The aim was to monitor cytokine levels in saliva and serum after locally fractionated radiotherapy of the head and neck in mice and investigate associations with salivary gland fibrosis and hyposalivation. C57BL\/6 mice were randomized to sham or X-ray irradiation of 66 Gy in 10 fractions over 5 days. Blood and saliva were collected on days \u22127, 5, 35, 80, and 105 following cytokine analysis. The harvested submandibular salivary gland was assessed for the presence of fibrosis. Decision tree regression analysis was used to investigate whether cytokine levels could predict late endpoints in terms of hyposalivation or fibrosis. Significant formation of fibrosis in gland tissue and reduced saliva production was found after irradiation. The pro-inflammatory cytokines IL-1\u03b1, TNF, TIMP1, G-CSF, KC, and MIP-1\u03b1 showed increased levels in saliva in irradiated mice and a strong correlation with late endpoints. The decision tree analysis largely separated controls from irradiated animals, with IL-1\u03b1 being the strongest predictor. Pro-inflammatory cytokines in saliva, but not in serum, were associated with late endpoints. This indicates that cytokine expression in saliva is a good biomarker for local salivary gland damage with IL-1\u03b1 as the strongest single predictor.<\/jats:p>","DOI":"10.3390\/ijms242015218","type":"journal-article","created":{"date-parts":[[2023,10,16]],"date-time":"2023-10-16T01:31:22Z","timestamp":1697419882000},"page":"15218","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Cytokine Levels in Saliva Are Associated with Salivary Gland Fibrosis and Hyposalivation in Mice after Fractionated Radiotherapy of the Head and Neck"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2744-9663","authenticated-orcid":false,"given":"Olga","family":"Zlygosteva","sequence":"first","affiliation":[{"name":"Department of Physics, University of Oslo, 0371 Oslo, Norway"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6317-5671","authenticated-orcid":false,"given":"Inga Solg\u00e5rd","family":"Juvkam","sequence":"additional","affiliation":[{"name":"Institute of Oral Biology, University of Oslo, 0372 Oslo, Norway"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hans Christian D.","family":"Aass","sequence":"additional","affiliation":[{"name":"The Blood Cell Research Group, Department of Medical Biochemistry, Oslo University Hospital, 0450 Oslo, Norway"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hilde K.","family":"Galtung","sequence":"additional","affiliation":[{"name":"Institute of Oral Biology, University of Oslo, 0372 Oslo, Norway"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tine M.","family":"S\u00f8land","sequence":"additional","affiliation":[{"name":"Institute of Oral Biology, University of Oslo, 0372 Oslo, Norway"},{"name":"Department of Pathology, Oslo University Hospital, 0372 Oslo, Norway"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eirik","family":"Malinen","sequence":"additional","affiliation":[{"name":"Department of Physics, University of Oslo, 0371 Oslo, Norway"},{"name":"Department of Radiation Biology, Oslo University Hospital, 0379 Oslo, Norway"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6995-131X","authenticated-orcid":false,"given":"Nina F. J.","family":"Edin","sequence":"additional","affiliation":[{"name":"Department of Physics, University of Oslo, 0371 Oslo, Norway"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,10,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1016\/j.oraloncology.2016.12.031","article-title":"Interventions for the management of radiotherapy-induced xerostomia and hyposalivation: A systematic review and meta-analysis","volume":"66","author":"Mercadante","year":"2017","journal-title":"Oral Oncol."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Jasmer, K.J., Gilman, K.E., Munoz Forti, K., Weisman, G.A., and Limesand, K.H. (2020). Radiation-Induced Salivary Gland Dysfunction: Mechanisms, Therapeutics and Future Directions. J. Clin. Med., 9.","DOI":"10.3390\/jcm9124095"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1667\/RR3031.1","article-title":"Cytokines in radiobiological responses: A review","volume":"178","author":"Schaue","year":"2012","journal-title":"Radiat. Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1186\/s12950-015-0058-3","article-title":"Portrait of inflammatory response to ionizing radiation treatment","volume":"12","author":"Minafra","year":"2015","journal-title":"J. Inflamm."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"574","DOI":"10.1080\/09553000902985136","article-title":"Milestones in normal tissue radiation biology over the past 50 years: From clonogenic cell survival to cytokine networks and back to stem cell recovery","volume":"85","author":"Stewart","year":"2009","journal-title":"Int. J. Radiat. Biol."},{"key":"ref_6","first-page":"491","article-title":"Macrophage cytokines: Involvement in immunity and infectious diseases","volume":"5","author":"Descoteaux","year":"2014","journal-title":"Front. Immunol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"367","DOI":"10.3389\/fonc.2019.00367","article-title":"Radiotherapy in Combination with Cytokine Treatment","volume":"9","author":"Palata","year":"2019","journal-title":"Front. Oncol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"103","DOI":"10.3857\/roj.2014.32.3.103","article-title":"Mechanisms of radiation-induced normal tissue toxicity and implications for future clinical trials","volume":"32","author":"Kim","year":"2014","journal-title":"Radiat. Oncol. J."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1093\/ajcp\/aqx095","article-title":"Robbins Basic Pathology","volume":"148","author":"Hoda","year":"2017","journal-title":"Am. J. Clin. Pathol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"647","DOI":"10.1002\/path.5389","article-title":"Radiation-induced tissue damage and response","volume":"250","author":"McBride","year":"2020","journal-title":"J. Pathol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"657","DOI":"10.1016\/j.clon.2012.04.001","article-title":"Radiation fibrosis\u2014Current clinical and therapeutic perspectives","volume":"24","author":"Westbury","year":"2012","journal-title":"Clin. Oncol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1985","DOI":"10.1007\/s00432-015-1974-6","article-title":"Radiation-induced fibrosis: Mechanisms and implications for therapy","volume":"141","author":"Straub","year":"2015","journal-title":"J. Cancer Res. Clin. Oncol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1046\/j.1365-2036.1996.22164021.x","article-title":"Cytokine assays and their limitations","volume":"10","author":"Tsang","year":"1996","journal-title":"Aliment. Pharmacol. Ther."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1016\/S1470-2045(03)01191-4","article-title":"Effects of radiation on normal tissue: Consequences and mechanisms","volume":"4","author":"Stone","year":"2003","journal-title":"Lancet Oncol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Gallet, P., Phulpin, B., Merlin, J.L., Leroux, A., Bravetti, P., Mecellem, H., Tran, N., and Dolivet, G. (2011). Long-term alterations of cytokines and growth factors expression in irradiated tissues and relation with histological severity scoring. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0029399"},{"key":"ref_16","first-page":"709","article-title":"Cytokines and radiation-induced pulmonary injuries","volume":"59","author":"Lierova","year":"2018","journal-title":"J. Radiat. Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1238","DOI":"10.1080\/09553002.2020.1807644","article-title":"Proton scanning and X-ray beam irradiation induce distinct regulation of inflammatory cytokines in a preclinical mouse model","volume":"96","author":"Nielsen","year":"2020","journal-title":"Int. J. Radiat. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Cunningham, S., McCauley, S., Vairamani, K., Speth, J., Girdhani, S., Abel, E., Sharma, R.A., Perentesis, J.P., Wells, S.I., and Mascia, A. (2021). FLASH Proton Pencil Beam Scanning Irradiation Minimizes Radiation-Induced Leg Contracture and Skin Toxicity in Mice. Cancers, 13.","DOI":"10.3390\/cancers13051012"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Ha, C.T., Li, X.H., Fu, D., Moroni, M., Fisher, C., Arnott, R., Srinivasan, V., and Xiao, M. (2014). Circulating interleukin-18 as a biomarker of total-body radiation exposure in mice, minipigs, and nonhuman primates (NHP). PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0109249"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1186\/1756-8722-2-6","article-title":"Radiation produces differential changes in cytokine profiles in radiation lung fibrosis sensitive and resistant mice","volume":"2","author":"Ao","year":"2009","journal-title":"J. Hematol. Oncol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1082","DOI":"10.1007\/s11764-022-01273-1","article-title":"Associations between patient-reported late effects and systemic cytokines in long-term survivors of head and neck cancer treated with radiotherapy","volume":"17","author":"Huynh","year":"2022","journal-title":"J. Cancer Surviv."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"499","DOI":"10.1007\/s00066-017-1119-8","article-title":"Early inflammatory changes in radiation-induced oral mucositis: Effect of pentoxifylline in a mouse model","volume":"193","author":"Gruber","year":"2017","journal-title":"Strahlenther. Onkol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"959","DOI":"10.1016\/j.ijrobp.2016.08.047","article-title":"Salivary Cytokine Levels and Oral Mucositis in Head and Neck Cancer Patients Treated with Chemotherapy and Radiation Therapy","volume":"96","author":"Bossi","year":"2016","journal-title":"Int. J. Radiat. Oncol. Biol. Phys."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"945","DOI":"10.1136\/ard.2009.115378","article-title":"Cytokines in Sjogren\u2019s syndrome: Potential therapeutic targets","volume":"69","author":"Roescher","year":"2010","journal-title":"Ann. Rheum. Dis."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1111\/odi.12252","article-title":"Saliva as a potential tool for diagnosis of dry mouth including Sjogren\u2019s syndrome","volume":"21","author":"Ohyama","year":"2015","journal-title":"Oral Dis."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1740","DOI":"10.1016\/j.cca.2011.06.026","article-title":"Salivary cytokines in cell proliferation and cancer","volume":"412","author":"Schapher","year":"2011","journal-title":"Clin. Chim. Acta"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1016\/j.oooo.2016.05.020","article-title":"Cytokines in saliva increase in head and neck cancer patients after treatment","volume":"122","author":"Russo","year":"2016","journal-title":"Oral Surg. Oral Med. Oral Pathol. Oral Radiol."},{"key":"ref_28","first-page":"970","article-title":"Salivary cytokine profiles in primary Sjogren\u2019s syndrome differ from those in non-Sjogren sicca in terms of TNF-alpha levels and Th-1\/Th-2 ratios","volume":"29","author":"Kang","year":"2011","journal-title":"Clin. Exp. Rheumatol."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Aqrawi, L.A., Chen, X., Hynne, H., Amdal, C., Reppe, S., Aass, H.C.D., Rykke, M., Hove, L.H., Young, A., and Herlofson, B.B. (2020). Cytokines Explored in Saliva and Tears from Radiated Cancer Patients Correlate with Clinical Manifestations, Influencing Important Immunoregulatory Cellular Pathways. Cells, 9.","DOI":"10.3390\/cells9092050"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"7319","DOI":"10.1038\/s41598-019-43714-5","article-title":"Elevated cytokine levels in tears and saliva of patients with primary Sj\u00f6gren\u2019s syndrome correlate with clinical ocular and oral manifestations","volume":"9","author":"Chen","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2673","DOI":"10.1158\/1055-9965.EPI-14-0588","article-title":"Usefulness of saliva samples for biomarker studies in radiation research","volume":"23","author":"Pernot","year":"2014","journal-title":"Cancer Epidemiol. Biomark. Prev."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"702","DOI":"10.1038\/nrc1950","article-title":"Preventing or reducing late side effects of radiation therapy: Radiobiology meets molecular pathology","volume":"6","author":"Bentzen","year":"2006","journal-title":"Nat. Rev. Cancer"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1016\/0360-3016(94)00421-G","article-title":"Late effects of radiation therapy in the head and neck region","volume":"31","author":"Cooper","year":"1995","journal-title":"Int. J. Radiat. Oncol. Biol. Phys."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"854","DOI":"10.1038\/s41419-020-03074-9","article-title":"Cellular senescence contributes to radiation-induced hyposalivation by affecting the stem\/progenitor cell niche","volume":"11","author":"Peng","year":"2020","journal-title":"Cell Death Dis."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Siva, S., MacManus, M., Kron, T., Best, N., Smith, J., Lobachevsky, P., Ball, D., and Martin, O. (2014). A pattern of early radiation-induced inflammatory cytokine expression is associated with lung toxicity in patients with non-small cell lung cancer. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0109560"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Yu, H.H., Chengchuan Ko, E., Chang, C.L., Yuan, K.S., Wu, A.T.H., Shan, Y.S., and Wu, S.Y. (2018). Fucoidan Inhibits Radiation-Induced Pneumonitis and Lung Fibrosis by Reducing Inflammatory Cytokine Expression in Lung Tissues. Mar. Drugs, 16.","DOI":"10.3390\/md16100392"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"779","DOI":"10.1093\/rheumatology\/37.7.779","article-title":"Chemokine expression and leucocyte infiltration in Sjogren\u2019s syndrome","volume":"37","author":"Cuello","year":"1998","journal-title":"Br. J. Rheumatol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2856518","DOI":"10.1155\/2018\/2856518","article-title":"Modulation of Inflammatory Reactions by Low-Dose Ionizing Radiation: Cytokine Release of Murine Endothelial Cells Is Dependent on Culture Conditions","volume":"2018","author":"Schroder","year":"2018","journal-title":"J. Immunol. Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1302","DOI":"10.4161\/cbt.6.8.4506","article-title":"Keratinocyte chemoattractant (KC)\/human growth-regulated oncogene (GRO) chemokines and pro-inflammatory chemokine networks in mouse and human ovarian epithelial cancer cells","volume":"6","author":"Son","year":"2007","journal-title":"Cancer Biol. Ther."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"7741","DOI":"10.1158\/1078-0432.CCR-08-1449","article-title":"Cytokine treatment improves parenchymal and vascular damage of salivary glands after irradiation","volume":"14","author":"Lombaert","year":"2008","journal-title":"Clin. Cancer Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1111\/j.1601-0825.2009.01582.x","article-title":"Cytokines in Sjogren\u2019s syndrome","volume":"15","author":"Roescher","year":"2009","journal-title":"Oral Dis."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"713","DOI":"10.1177\/0022034519837240","article-title":"Targeted TNF-alpha Overexpression Drives Salivary Gland Inflammation","volume":"98","author":"Limaye","year":"2019","journal-title":"J. Dent. Res."},{"key":"ref_43","unstructured":"Bhavsar, I., Miller, C.S., and Al-Sabbagh, M. (2015). General Methods in Biomarker Research and Their Applications, Springer."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"S145","DOI":"10.1016\/j.ijrobp.2009.08.076","article-title":"Biomarkers and surrogate endpoints for normal-tissue effects of radiation therapy: The importance of dose-volume effects","volume":"76","author":"Bentzen","year":"2010","journal-title":"Int. J. Radiat. Oncol. Biol. Phys."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1093\/jrr\/rrac066","article-title":"A preclinical model to investigate normal tissue damage following fractionated radiotherapy to the head and neck","volume":"64","author":"Juvkam","year":"2022","journal-title":"J. Radiat. Res."},{"key":"ref_46","first-page":"e57203","article-title":"A Method for the Measurement of Salivary Gland Function in Mice","volume":"131","author":"Bagavant","year":"2018","journal-title":"J. Vis. Exp."},{"key":"ref_47","first-page":"2825","article-title":"Scikit-learn: Machine learning in python","volume":"12","author":"Pedregosa","year":"2011","journal-title":"J. Mach. Learn. Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/20\/15218\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:07:26Z","timestamp":1760130446000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/20\/15218"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,10,16]]},"references-count":47,"journal-issue":{"issue":"20","published-online":{"date-parts":[[2023,10]]}},"alternative-id":["ijms242015218"],"URL":"https:\/\/doi.org\/10.3390\/ijms242015218","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,10,16]]}}}