Utilizing a naturopathic mouthwash with selective antimicrobial effects against multispecies oral biofilms for prevention of dysbiosis

IntroductionOral 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 o...

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Main Authors: Danyal A. Siddiqui, Yi-Wen C. Tsai, Juliana Giron Bastidas, Marzieh S. Jazaeri, Georgios A. Kotsakis
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-05-01
Series:Frontiers in Oral Health
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Online Access:https://www.frontiersin.org/articles/10.3389/froh.2025.1529061/full
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author Danyal A. Siddiqui
Danyal A. Siddiqui
Yi-Wen C. Tsai
Yi-Wen C. Tsai
Juliana Giron Bastidas
Juliana Giron Bastidas
Marzieh S. Jazaeri
Marzieh S. Jazaeri
Georgios A. Kotsakis
Georgios A. Kotsakis
author_facet Danyal A. Siddiqui
Danyal A. Siddiqui
Yi-Wen C. Tsai
Yi-Wen C. Tsai
Juliana Giron Bastidas
Juliana Giron Bastidas
Marzieh S. Jazaeri
Marzieh S. Jazaeri
Georgios A. Kotsakis
Georgios A. Kotsakis
author_sort Danyal A. Siddiqui
collection DOAJ
description IntroductionOral 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.MethodsStreptoccoccus oralis, Streptococcus gordonii, Veillonella parvula, Fusobacterium nucleatum, and Porphyromonas gingivalis were cultured and treated with naturopathic StellaLife® VEGA® Oral Rinse (SL), 0.12% chlorhexidine gluconate (CHX), LISTERINE® COOL MINT® 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.ResultsSL significantly inhibited the growth of disease-associated F. nucleatum and P. gingivalis 12 and 120 h, respectively, after treatment, while exhibiting lower toxicity toward commensal S. oralis, S. gordonii, and V. parvula vs. LIS or CHX (all p < 0.05). Correspondingly, in 4-species biofilms, selective agar plating and FISH-staining showed decreased abundance of F. nucleatum and P. gingivalis after 4 h recovery following SL treatment vs. PBS control while maintaining a robust commensal biofilm of S. oralis and V. parvula. In contrast, CHX or LIS treatment demonstrated non-selective killing, leading to sparse biofilms with residual F. nucleatum and P. gingivalis. When tested against clinical ex vivo multispecies biofilms, all oral rinses showed significant antibiofilm effects (all p < 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.DiscussionThe 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.
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spelling doaj-art-dac8f79cf87d4d97a9f27ff89742ca952025-08-20T03:53:46ZengFrontiers Media S.A.Frontiers in Oral Health2673-48422025-05-01610.3389/froh.2025.15290611529061Utilizing a naturopathic mouthwash with selective antimicrobial effects against multispecies oral biofilms for prevention of dysbiosisDanyal A. Siddiqui0Danyal A. Siddiqui1Yi-Wen C. Tsai2Yi-Wen C. Tsai3Juliana Giron Bastidas4Juliana Giron Bastidas5Marzieh S. Jazaeri6Marzieh S. Jazaeri7Georgios A. Kotsakis8Georgios A. Kotsakis9Department of Oral Biology, Rutgers School of Dental Medicine, Newark, NJ, United StatesClinical Research Center, Rutgers School of Dental Medicine, Newark, NJ, United StatesTranslational Periodontal Research Lab, Department of Periodontics, University of Texas Health Science Center at San Antonio, San Antonio, TX, United StatesDepartment of Periodontology, School of Dentistry, Tri-Service General Hospital and National Defense Medical Center, Taipei, TaiwanDepartment of Oral Biology, Rutgers School of Dental Medicine, Newark, NJ, United StatesClinical Research Center, Rutgers School of Dental Medicine, Newark, NJ, United StatesDepartment of Oral Biology, Rutgers School of Dental Medicine, Newark, NJ, United StatesClinical Research Center, Rutgers School of Dental Medicine, Newark, NJ, United StatesDepartment of Oral Biology, Rutgers School of Dental Medicine, Newark, NJ, United StatesClinical Research Center, Rutgers School of Dental Medicine, Newark, NJ, United StatesIntroductionOral 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.MethodsStreptoccoccus oralis, Streptococcus gordonii, Veillonella parvula, Fusobacterium nucleatum, and Porphyromonas gingivalis were cultured and treated with naturopathic StellaLife® VEGA® Oral Rinse (SL), 0.12% chlorhexidine gluconate (CHX), LISTERINE® COOL MINT® 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.ResultsSL significantly inhibited the growth of disease-associated F. nucleatum and P. gingivalis 12 and 120 h, respectively, after treatment, while exhibiting lower toxicity toward commensal S. oralis, S. gordonii, and V. parvula vs. LIS or CHX (all p < 0.05). Correspondingly, in 4-species biofilms, selective agar plating and FISH-staining showed decreased abundance of F. nucleatum and P. gingivalis after 4 h recovery following SL treatment vs. PBS control while maintaining a robust commensal biofilm of S. oralis and V. parvula. In contrast, CHX or LIS treatment demonstrated non-selective killing, leading to sparse biofilms with residual F. nucleatum and P. gingivalis. When tested against clinical ex vivo multispecies biofilms, all oral rinses showed significant antibiofilm effects (all p < 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.DiscussionThe 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.https://www.frontiersin.org/articles/10.3389/froh.2025.1529061/fulloral rinseschlorhexidineantimicrobial(s)oral biofilmsperiodontitisgingivitis
spellingShingle Danyal A. Siddiqui
Danyal A. Siddiqui
Yi-Wen C. Tsai
Yi-Wen C. Tsai
Juliana Giron Bastidas
Juliana Giron Bastidas
Marzieh S. Jazaeri
Marzieh S. Jazaeri
Georgios A. Kotsakis
Georgios A. Kotsakis
Utilizing a naturopathic mouthwash with selective antimicrobial effects against multispecies oral biofilms for prevention of dysbiosis
Frontiers in Oral Health
oral rinses
chlorhexidine
antimicrobial(s)
oral biofilms
periodontitis
gingivitis
title Utilizing a naturopathic mouthwash with selective antimicrobial effects against multispecies oral biofilms for prevention of dysbiosis
title_full Utilizing a naturopathic mouthwash with selective antimicrobial effects against multispecies oral biofilms for prevention of dysbiosis
title_fullStr Utilizing a naturopathic mouthwash with selective antimicrobial effects against multispecies oral biofilms for prevention of dysbiosis
title_full_unstemmed Utilizing a naturopathic mouthwash with selective antimicrobial effects against multispecies oral biofilms for prevention of dysbiosis
title_short Utilizing a naturopathic mouthwash with selective antimicrobial effects against multispecies oral biofilms for prevention of dysbiosis
title_sort utilizing a naturopathic mouthwash with selective antimicrobial effects against multispecies oral biofilms for prevention of dysbiosis
topic oral rinses
chlorhexidine
antimicrobial(s)
oral biofilms
periodontitis
gingivitis
url https://www.frontiersin.org/articles/10.3389/froh.2025.1529061/full
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