{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T11:01:58Z","timestamp":1783594918453,"version":"3.55.0"},"reference-count":39,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,5,19]],"date-time":"2025-05-19T00:00:00Z","timestamp":1747612800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Oral Health"],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>Oral rinses intended for the prevention and treatment of periodontal diseases have traditionally focused on bactericidal effects. This study evaluates the efficacy of a naturopathic mouthwash containing plant attenuations and propolis against common gram- pathogenic and gram+ commensal oral species in comparison to conventional antiseptic oral rinses.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p><jats:italic>Streptoccoccus oralis, Streptococcus gordonii, Veillonella parvula, Fusobacterium nucleatum, and Porphyromonas gingivalis<\/jats:italic> were cultured and treated with naturopathic StellaLife\u00ae VEGA\u00ae Oral Rinse (SL), 0.12% chlorhexidine gluconate (CHX), LISTERINE\u00ae COOL MINT\u00ae mouthwash (LIS), or phosphate-buffered saline (PBS) as negative control. Firstly, planktonic bacterial growth was assessed through optical density measurements and colony-forming unit (CFU) counts. Subsequently, a 4-species or clinical ex vivo multispecies biofilm was used to evaluate antibiofilm effects through selective agar plating and fluorescence in situ hybridization (FISH) or live-dead biofilm imaging, respectively. Lastly, cytocompatibility to oral rinses was tested using a 3D human fibroblast spheroid model.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>SL significantly inhibited the growth of disease-associated <jats:italic>F. nucleatum and P. gingivalis<\/jats:italic> 12 and 120 h, respectively, after treatment, while exhibiting lower toxicity toward commensal <jats:italic>S. oralis, S. gordonii<\/jats:italic>, and <jats:italic>V. parvula<\/jats:italic> vs. <jats:italic>LIS<\/jats:italic> or CHX (all <jats:italic>p<\/jats:italic> &amp;lt; 0.05). Correspondingly, in 4-species biofilms, selective agar plating and FISH-staining showed decreased abundance of <jats:italic>F. nucleatum and P. gingivalis<\/jats:italic> after 4 h recovery following SL treatment vs. PBS control while maintaining a robust commensal biofilm of <jats:italic>S. oralis and V. parvula<\/jats:italic>. In contrast, CHX or LIS treatment demonstrated non-selective killing, leading to sparse biofilms with residual <jats:italic>F. nucleatum and P. gingivalis<\/jats:italic>. When tested against clinical ex vivo multispecies biofilms, all oral rinses showed significant antibiofilm effects (all <jats:italic>p<\/jats:italic> &amp;lt; 0.001), disrupting biofilm structure and reducing bacterial viability. Lastly, 3D human fibroblast spheroids treated with CHX or LIS displayed greater cytotoxicity with detachment of cellular debris from the spheroid mass, while spheroids exposed to SL exhibited minimal cell death with cellular viability maintained across the spheroid structure.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>The SL homeopathic rinse demonstrated selective action on oral bacteria, preferentially reducing pathogen bacterial load while preserving commensal species with high cytocompatibility. Future validation in human studies is needed to assess its selective antimicrobial activity to maintain a eubiotic oral microbiome and explore broader applications in oral health.<\/jats:p><\/jats:sec>","DOI":"10.3389\/froh.2025.1529061","type":"journal-article","created":{"date-parts":[[2025,5,19]],"date-time":"2025-05-19T05:14:54Z","timestamp":1747631694000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":4,"title":["Utilizing a naturopathic mouthwash with selective antimicrobial effects against multispecies oral biofilms for prevention of dysbiosis"],"prefix":"10.3389","volume":"6","author":[{"given":"Danyal A.","family":"Siddiqui","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi-Wen C.","family":"Tsai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juliana","family":"Giron Bastidas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marzieh S.","family":"Jazaeri","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Georgios A.","family":"Kotsakis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,5,19]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"103183","DOI":"10.1016\/j.jksus.2024.103183","article-title":"A meta-analysis of cohort and pilot studies indicates association of mouthwash use and risk of hypertension in adults","volume":"36","author":"Bagabir","year":"2024","journal-title":"J King Saud Univ Sci"},{"key":"B2","doi-asserted-by":"publisher","first-page":"S74","DOI":"10.1016\/j.identj.2023.08.010","article-title":"Mouthwash effects on the oral microbiome: are they good, bad, or balanced?","volume":"73","author":"Brookes","year":"2023","journal-title":"Int Dent J"},{"key":"B3","doi-asserted-by":"publisher","first-page":"103","DOI":"10.1080\/08037051.2019.1680270","article-title":"Over-the-counter mouthwash use, nitric oxide and hypertension risk","volume":"29","author":"Joshipura","year":"2020","journal-title":"Blood Press"},{"key":"B4","doi-asserted-by":"publisher","first-page":"333","DOI":"10.1016\/j.niox.2008.08.003","article-title":"The increase in plasma nitrite after a dietary nitrate load is markedly attenuated by an antibacterial mouthwash","volume":"19","author":"Govoni","year":"2008","journal-title":"Nitric Oxide"},{"key":"B5","doi-asserted-by":"publisher","first-page":"461","DOI":"10.4103\/0972-0707.168810","article-title":"Antibacterial efficacy of Azadirachta indica, Mimusops elengi and 2% CHX on multispecies dentinal biofilm","volume":"18","author":"Mistry","year":"2015","journal-title":"J Conserv Dent"},{"key":"B6","doi-asserted-by":"publisher","first-page":"172","DOI":"10.4103\/1305-7456.130591","article-title":"The antimicrobial activity of Azadirachta indica, Mimusops elengi, Tinospora cardifolia, Ocimum sanctum and 2% chlorhexidine gluconate on common endodontic pathogens: an in vitro study","volume":"8","author":"Mistry","year":"2014","journal-title":"Eur J Dent"},{"key":"B7","doi-asserted-by":"publisher","first-page":"15","DOI":"10.4103\/0970-9290.179808","article-title":"Efficacy of 10% whole Azadirachta indica (neem) chip as an adjunct to scaling and root planning in chronic periodontitis: a clinical and microbiological study","volume":"27","author":"Vennila","year":"2016","journal-title":"Indian J Dent Res"},{"key":"B8","doi-asserted-by":"publisher","first-page":"1205","DOI":"10.1099\/jmm.0.000343","article-title":"Intracellular, biofilm-inhibitory and membrane-damaging activities of nimbolide isolated from Azadirachta indica A. 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