{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T14:57:40Z","timestamp":1791298660705,"version":"4.1.0"},"reference-count":52,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2023,6,9]],"date-time":"2023-06-09T00:00:00Z","timestamp":1686268800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Croatian Science Foundation within the framework of the Slovenian","award":["IPS-2020-01-7418"],"award-info":[{"award-number":["IPS-2020-01-7418"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The mechanism of toxicity and cellular response to metal ions present in the environment is still a very current area of research. In this work, which is a continuation of the study of the toxicity of metal ions released by fixed orthodontic appliances, eluates of archwires, brackets, ligatures, and bands are used to test the prooxidant effect, cytotoxicity, and genotoxicity on cell lines of the gastrointestinal tract. Eluates obtained after three immersion periods (3, 7, and 14 days) and with known amounts and types of metal ions were used. Four cell lines\u2014CAL 27 (human tongue), Hep-G2 (liver), AGS (stomach) and CaCo-2 (colon)\u2014were treated with each type of eluate at four concentrations (0.1\u00d7, 0.5\u00d7, 1.0\u00d7, and 2.0\u00d7) for 24 h. Most eluates had toxic effects on CAL 27 cells over the entire concentration range regardless of exposure time, while CaCo-2 proved to be the most resistant. In AGS and Hep-G2 cells, all samples tested induced free radical formation, with the highest concentration (2\u00d7) causing a decrease in free radicals formed compared to the lowest concentrations. Eluates containing Cr, Mn, and Al showed a slight pro-oxidant effect on DNA (on plasmid \u03c6X-174 RF I) and slight genotoxicity (comet assay), but these effects are not so great that the human body could not \u201cresist\u201d them. Statistical analysis of data on chemical composition, cytotoxicity, ROS, genotoxicity, and prooxidative DNA damage shows the influence of metal ions present in some eluates on the toxicity obtained. Fe and Ni are responsible for the production of ROS, while Mn and Cr have a great influence on hydroxyl radicals, which cause single-strand breaks in supercoiled plasmid DNA in addition to the production of ROS. On the other hand, Fe, Cr, Mn, and Al are responsible for the cytotoxic effect of the studied eluates. The obtained results confirm that this type of research is useful and brings us closer to more accurate in vivo conditions.<\/jats:p>","DOI":"10.3390\/ijms24129940","type":"journal-article","created":{"date-parts":[[2023,6,9]],"date-time":"2023-06-09T03:30:03Z","timestamp":1686281403000},"page":"9940","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["Toxicity of Metal Ions Released from a Fixed Orthodontic Appliance to Gastrointestinal Tract Cell Lines"],"prefix":"10.3390","volume":"24","author":[{"given":"Ksenija","family":"Durgo","sequence":"first","affiliation":[{"name":"Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, Universtiy of Zagreb, Pierrotijeva 6, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7529-7740","authenticated-orcid":false,"given":"Sun\u010dana","family":"Ore\u0161i\u0107","sequence":"additional","affiliation":[{"name":"Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, Universtiy of Zagreb, Pierrotijeva 6, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-6056-5334","authenticated-orcid":false,"given":"Marijana","family":"Rin\u010di\u0107 Mlinari\u0107","sequence":"additional","affiliation":[{"name":"Private Orthodontic Practice, Katarine Zrinske 1b, 23000 Zadar, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7628-9045","authenticated-orcid":false,"given":"\u017deljka","family":"Fiket","sequence":"additional","affiliation":[{"name":"Division for Marine and Environmental Research, Ru\u0111er Bo\u0161kovi\u0107 Institute, Bijeni\u010dka 54, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gordana \u010canadi","family":"Jure\u0161i\u0107","sequence":"additional","affiliation":[{"name":"Department of Medical Chemistry, Biochemistry and Clinical Chemistry, Faculty of Medicine, Universtiy of Rijeka, B. Branchetta 20, 51000 Rijeka, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,6,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/j.etap.2015.05.007","article-title":"In Vitro and in Vivo Evidence of the Cytotoxic and Genotoxic Effects of Metal Ions Released by Orthodontic Appliances: A Review","volume":"40","author":"Jos","year":"2015","journal-title":"Environ. Toxicol. Pharmacol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"494","DOI":"10.1007\/s00056-017-0107-z","article-title":"Metal Release Profiles of Orthodontic Bands, Brackets, and Wires: An in Vitro Study","volume":"78","author":"Wendl","year":"2017","journal-title":"J. Orofac. Orthop."},{"key":"ref_3","unstructured":"Brantley, W., Berzins, D., Iijima, M., Tufek\u00e7i, E., and Cai, Z. (2017). Orthodontic Applications of Biomaterials, Elsevier."},{"key":"ref_4","unstructured":"Proffit, W.R. (2018). Contemporary Orthodontics, Elsevier. [6th ed.]."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1159\/000261239","article-title":"Effect of Dental Plaque Age and Bacterial Composition on the PH of Artificial Fissures in Human Volunteers","volume":"24","author":"Igarashi","year":"1990","journal-title":"Caries Res."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Ferrer, M.D., P\u00e9rez, S., Lopez, A.L., Sanz, J.L., Melo, M., Llena, C., and Mira, A. (2021). Evaluation of Clinical, Biochemical and Microbiological Markers Related to Dental Caries. Int. J. Environ. Res. Public Health, 18.","DOI":"10.3390\/ijerph18116049"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"440","DOI":"10.3109\/15376516.2015.1062951","article-title":"Genotoxic and Cytotoxic Effects and Gene Expression Changes Induced by Fixed Orthodontic Appliances in Oral Mucosa Cells of Patients: A Systematic Review","volume":"25","author":"Jos","year":"2015","journal-title":"Toxicol. Mech. Methods"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1007\/s12011-009-8576-6","article-title":"Trace Metal Release from Orthodontic Appliances by In Vivo Studies: A Systematic Literature Review","volume":"137","author":"Mikulewicz","year":"2010","journal-title":"Biol. Trace Elem. Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"957","DOI":"10.1016\/j.ajodo.2016.10.030","article-title":"Cytotoxic Outcomes of Orthodontic Bands with and without Silver Solder in Different Cell Lineages","volume":"151","author":"Jacoby","year":"2017","journal-title":"Am. J. Orthod. Dentofac. Orthop."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"500","DOI":"10.1016\/j.ortho.2021.04.006","article-title":"In Vitro Evaluation of Immediate Cytotoxicity of Resterilised Orthodontic Bands on HGF-1 Cell Line","volume":"19","author":"Ajami","year":"2021","journal-title":"Int. Orthod."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"349","DOI":"10.2319\/050620-405.1","article-title":"Cytotoxic Evaluation of Two Orthodontic Silver Solder Materials on Human Periodontal Ligament Fibroblast Cells and the Effects of Antioxidant and Antiapoptotic Reagents","volume":"91","author":"Nimeri","year":"2021","journal-title":"Angle Orthod."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Wishney, M., Mahadevan, S., Cornwell, J.A., Savage, T., Proschogo, N., Darendeliler, M.A., and Zoellner, H. (2022). Toxicity of Orthodontic Brackets Examined by Single Cell Tracking. Toxics, 10.","DOI":"10.3390\/toxics10080460"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"8885290","DOI":"10.1155\/2021\/8885290","article-title":"Comparison of Cell Viability and Chemical Composition of Six Latest Generation Orthodontic Wires","volume":"2021","author":"Escobar","year":"2021","journal-title":"Int. J. Biomater."},{"key":"ref_14","first-page":"1467456","article-title":"In Vitro Determination of Genotoxicity Induced by Brackets Alloys in Cultures of Human Gingival Fibroblasts","volume":"2020","year":"2020","journal-title":"J. Toxicol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"114784","DOI":"10.1016\/j.taap.2019.114784","article-title":"Cytotoxicity and Oxidative Stress Induced by Nickel and Titanium Ions from Dental Alloys on Cells of Gastrointestinal Tract","volume":"383","author":"Durgo","year":"2019","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.mrgentox.2014.01.011","article-title":"Cytotoxicity and Genotoxicity of Orthodontic Bands with or without Silver Soldered Joints","volume":"762","author":"Menezes","year":"2014","journal-title":"Mutat. Res. Genet. Toxicol. Environ. Mutagen."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Kova\u010d, V., Polj\u0161ak, B., Primo\u017ei\u010d, J., and Jamnik, P. (2020). Are Metal Ions That Make up Orthodontic Alloys Cytotoxic, and Do They Induce Oxidative Stress in a Yeast Cell Model?. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21217993"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Kova\u010d, V., Bergant, M., \u0160\u010dan\u010dar, J., Primo\u017ei\u010d, J., Jamnik, P., and Polj\u0161ak, B. (2021). Causation of Oxidative Stress and Defense Response of a Yeast Cell Model after Treatment with Orthodontic Alloys Consisting of Metal Ions. Antioxidants, 11.","DOI":"10.3390\/antiox11010063"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Papaioannou, P., S\u00fctel, M., H\u00fcsker, K., M\u00fcller, W.-D., and Bartzela, T. (2021). A New Setup for Simulating the Corrosion Behavior of Orthodontic Wires. Materials, 14.","DOI":"10.3390\/ma14133758"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/j.jobcr.2016.06.001","article-title":"Nickel Release from Stainless Steel and Nickel Titanium Archwires\u2014An in Vitro Study","volume":"6","author":"Hussain","year":"2016","journal-title":"J. Oral Biol. Craniofacial Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"11","DOI":"10.4103\/jos.jos_157_21","article-title":"Ion Release and Surface Roughness of Silver Soldered Bands with Two Different Polishing Methods: An in-Vitro Study","volume":"11","year":"2022","journal-title":"J. Orthodont. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Petkovi\u0107 Didovi\u0107, M., Jelovica Badovinac, I., Fiket, \u017d., \u017digon, J., Rin\u010di\u0107 Mlinari\u0107, M., and \u010canadi Jure\u0161i\u0107, G. (2023). Cytotoxicity of Metal Ions Released from NiTi and Stainless Steel Orthodontic Appliances, Part 1: Surface Morphology and Ion Release Variations. Materials, 16.","DOI":"10.3390\/ma16114156"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"102","DOI":"10.2319\/083007-401.1","article-title":"Type of Archwire and Level of Acidity: Effects on the Release of Metal Ions from Orthodontic Appliances","volume":"79","author":"Kuhta","year":"2009","journal-title":"Angle Orthod."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1067\/mod.2001.117911","article-title":"Metal Release from Simulated Fixed Orthodontic Appliances","volume":"120","author":"Hwang","year":"2001","journal-title":"Am. J. Orthod. Dentofac. Orthop."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Hsu, M.Y., Mina, E., Roetto, A., and Porporato, P.E. (2020). Iron: An Essential Element of Cancer Metabolism. Cells, 9.","DOI":"10.3390\/cells9122591"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"308","DOI":"10.1093\/jas\/skab235.565","article-title":"PSV-13 Understanding the Role of Zinc and Manganese in Proliferation and Protein Synthesis of Primary Bovine Satellite Cells","volume":"99","author":"Alberto","year":"2021","journal-title":"J. Anim. Sci."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Francisco, L.F.V., Baldivia, D.D.S., Crispim, B.D.A., Klafke, S.M.F.F., Castilho, P.F.D., Viana, L.F., Santos, E.L.D., Oliveira, K.M.P.D., and Barufatti, A. (2021). Acute Toxic and Genotoxic Effects of Aluminum and Manganese Using In Vitro Models. Toxics, 9.","DOI":"10.3390\/toxics9070153"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"590226","DOI":"10.3389\/fcell.2020.590226","article-title":"Iron Metabolism in Ferroptosis","volume":"8","author":"Chen","year":"2020","journal-title":"Front. Cell Dev. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Primo\u017ei\u010d, J., Polj\u0161ak, B., Jamnik, P., Kova\u010d, V., \u010canadi Jure\u0161i\u0107, G., and Spalj, S. (2021). Risk Assessment of Oxidative Stress Induced by Metal Ions Released from Fixed Orthodontic Appliances during Treatment and Indications for Supportive Antioxidant Therapy: A Narrative Review. Antioxidants, 10.","DOI":"10.3390\/antiox10091359"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Aliofkhazraei, M. (2015). Superalloys, IntechOpen.","DOI":"10.5772\/59358"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1155","DOI":"10.17219\/acem\/65726","article-title":"Trace Metal Ions Release from Fixed Orthodontic Appliances and DNAdamage in Oral Mucosa Cells by in Vivo Studies: A Literature Review","volume":"26","author":"Downarowicz","year":"2017","journal-title":"Adv. Clin. Exp. Med."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1177\/154405910708600510","article-title":"In Vitro Evaluation of Corrosion and Cytotoxicity of Orthodontic Brackets","volume":"86","author":"Costa","year":"2007","journal-title":"J. Dent. Res."},{"key":"ref_33","first-page":"212","article-title":"[In Vitro Cytotoxicity Study of Nickel Ion]","volume":"39","author":"Wen","year":"2015","journal-title":"Zhongguo Yi Liao Qi Xie Za Zhi"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1186\/s13018-014-0091-6","article-title":"Cytotoxic Effects of Cobalt and Nickel Ions on Osteocytes in Vitro","volume":"9","author":"Kanaji","year":"2014","journal-title":"J. Orthop. Surg. Res."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Wu, Y., Lin, J., Wang, T., Lin, T., Yen, M., Liu, Y., Wu, P., Chen, F., Shih, Y., and Yeh, I. (2019). Hexavalent Chromium Intoxication Induces Intrinsic and Extrinsic Apoptosis in Human Renal Cells. Mol. Med. Rep., 851\u2013857.","DOI":"10.3892\/mmr.2019.10885"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1517\/17425255.1.2.175","article-title":"Caco-2 Cell Permeability Assays to Measure Drug Absorption","volume":"1","author":"Li","year":"2005","journal-title":"Expert Opin. Drug Metab. Toxicol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1093\/oxfordjournals.rpd.a006746","article-title":"The Production of Single Strand and Double Strand Breaks in DNA in Aqueous Solution by Vacuum UV Photons Below 10 Ev","volume":"99","author":"Folkard","year":"2002","journal-title":"Radiat. Prot. Dosim."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"881","DOI":"10.1080\/09553000050050891","article-title":"Critical Energies for SSB and DSB Induction in Plasmid DNA by Low-Energy Photons: Action Spectra for Strand-Break Induction in Plasmid DNA Irradiated in Vacuum","volume":"76","author":"Prise","year":"2000","journal-title":"Int. J. Radiat. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1006\/jmbi.2001.5031","article-title":"Electron and Scanning Force Microscopy Studies of Alterations in Supercoiled DNA Tertiary Structure","volume":"313","author":"Cherny","year":"2001","journal-title":"J. Mol. Biol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"609","DOI":"10.1146\/annurev.bb.23.060194.003141","article-title":"Conformational and Thermodynamic Properties of Supercoiled DNA","volume":"23","author":"Vologodskii","year":"1994","journal-title":"Annu. Rev. Biophys. Biomol. Struct."},{"key":"ref_41","first-page":"57","article-title":"Mutagenesis by Metal-Induced Oxygen Radicals","volume":"102","author":"Reid","year":"1994","journal-title":"Environ. Health Perspect."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1358","DOI":"10.1039\/C4MT00057A","article-title":"Metal-Mediated DNA Damage and Cell Death: Mechanisms, Detection Methods, and Cellular Consequences","volume":"6","author":"Goodman","year":"2014","journal-title":"Metallomics"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"298","DOI":"10.1016\/j.ajodo.2010.05.025","article-title":"Cytotoxicity, Genotoxicity, and Metal Release in Patients with Fixed Orthodontic Appliances: A Longitudinal in-Vivo Study","volume":"140","author":"Hafez","year":"2011","journal-title":"Am. J. Orthod. Dentofac. Orthop."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"97","DOI":"10.18585\/inabj.v8i2.193","article-title":"The Release of Total Metal Ion and Genotoxicity of Stainless Steel Brackets: Experimental Study Using Micronucleus Assay","volume":"8","author":"Karlina","year":"2016","journal-title":"Indones. Biomed. J."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1539","DOI":"10.14260\/jemds\/2021\/321","article-title":"Evaluation of Nickel Release in Blood and Periodontal Tissue with the Use of NiTi Wires, Bands and Brackets in Orthodontics\u2013A Systematic Review","volume":"10","author":"Kishore","year":"2021","journal-title":"J. Evol. Med. Dent. Sci."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.cotox.2019.05.003","article-title":"Mechanisms of Chromium-Induced Toxicity","volume":"14","author":"DesMarias","year":"2019","journal-title":"Curr. Opin. Toxicol."},{"key":"ref_47","unstructured":"(2009). Biological Evaluation of Medical Devices\u2014Part 5: Tests for In Vitro Cytotoxicity (Standard No. ISO 10993-5:2009)."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1125","DOI":"10.1038\/nprot.2008.75","article-title":"Neutral Red Uptake Assay for the Estimation of Cell Viability\/Cytotoxicity","volume":"3","author":"Repetto","year":"2008","journal-title":"Nat. Protoc."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"3019","DOI":"10.1039\/C4AY00288A","article-title":"Measurement of DCF Fluorescence as a Measure of Reactive Oxygen Species in Murine Islets of Langerhans","volume":"6","author":"Wang","year":"2014","journal-title":"Anal. Methods"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"pdb-top100388","DOI":"10.1101\/pdb.top100388","article-title":"Analysis of DNA by Agarose Gel Electrophoresis","volume":"2019","author":"Green","year":"2019","journal-title":"Cold Spring Harb. Protoc."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1007\/s11010-011-1206-6","article-title":"EPR Studies on Hydroxyl Radical-Scavenging Activities of Pravastatin and Fluvastatin","volume":"364","author":"Vandjelovic","year":"2012","journal-title":"Mol. Cell Biochem."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"949","DOI":"10.1007\/s00204-013-1070-0","article-title":"The Essential Comet Assay: A Comprehensive Guide to Measuring DNA Damage and Repair","volume":"87","author":"Azqueta","year":"2013","journal-title":"Arch. Toxicol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/12\/9940\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T19:51:26Z","timestamp":1760125886000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/12\/9940"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,6,9]]},"references-count":52,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2023,6]]}},"alternative-id":["ijms24129940"],"URL":"https:\/\/doi.org\/10.3390\/ijms24129940","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,6,9]]}}}