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A 16-h culture of <jats:italic>S. mutans<\/jats:italic> UA159 was treated with the mouthwashes in three dilutions (1:3, 1:6, and 1:12), prepared in Tryptic Soy broth supplemented with 1% sucrose. The minimum inhibitory concentrations (MIC), planktonic, and biofilm growth were evaluated using a spectrophotometer. In addition, the growth for minimum bactericidal concentration (MBC) was evaluated using five \u03bcL of the dilution and incubated on blood agar. For the analysis of the results, the mouthwashes were separated into six groups according to their active ingredients (cetylpyridinium chloride\/CPC, n\u2009=\u200925; essential oils\/EO n\u2009=\u200910; whitening\/W (hydrogen peroxide\/ sodium hexametaphosphate), n\u2009=\u200912; Natural-Derived Actives \/ NDA, n\u2009=\u200915; zinc chloride\/ZC, n\u2009=\u20093; others\/O, n\u2009=\u200916). ANOVA followed by the Tukey test was performed (p\u2009&lt;\u20090.05).<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>Regarding MIC, planktonic, and biofilm growth of <jats:italic>S. mutans<\/jats:italic>, there was a significant decrease for the W and CPC groups (p\u2009&lt;\u20090.001). The EO and W groups had more inhibition on <jats:italic>S. mutans<\/jats:italic> biofilm compared to the CPC group (p\u2009&lt;\u20090.05). For ZC, NDA, and O groups, there were different effects within the same group, presenting a large variability. About MBC, W and CPC groups presented the higher inhibition (W\u2009&gt;\u2009CPC\u2009&gt;\u2009EO\u2009&gt;\u2009NDA\/ZC\/O).<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusion<\/jats:title>\n            <jats:p>The mouthwashes demonstrated significant effect on S. <jats:italic>mutans<\/jats:italic> biofilm, especially in the 1:3 dilution. W and CPC groups had a more significant effect on <jats:italic>S. mutans<\/jats:italic> biofilm.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Clinical relevance<\/jats:title>\n            <jats:p>\n              <jats:italic>S. mutans<\/jats:italic> is an important bacterium in dental caries and periodontal diseases. Our study showed that non-fluoridated mouthwashes affect the initial stages of biofilm formation.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1007\/s00784-025-06462-7","type":"journal-article","created":{"date-parts":[[2025,8,14]],"date-time":"2025-08-14T04:23:48Z","timestamp":1755145428000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["In vitro effect of fluoride-free mouthwashes on Streptococcus mutans biofilm"],"prefix":"10.1007","volume":"29","author":[{"given":"Astrid C.","family":"Valdivia-Tapia","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Frank","family":"Lippert","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter F.","family":"Castelluccio","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jaime A.","family":"Cury","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antonio P.","family":"Ricomini-Filho","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Richard L.","family":"Gregory","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2025,8,14]]},"reference":[{"key":"6462_CR1","doi-asserted-by":"publisher","first-page":"3926","DOI":"10.3390\/ijerph19073926","volume":"19","author":"D Radzki","year":"2022","unstructured":"Radzki D, Wilhelm-W\u0119glarz M, Pruska K, Kusiak A, Ordyniec-Kwa\u015bnica I (2022) A fresh look at mouthwashes\u2014what is inside and what is it for? 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