An interventional study of Essential oil mouthwash and Chlorhexidine gluconate alcohol-free mouthwash in Oral Hygiene, Oral Health, Dental Plaque and Respiratory Tract Infections (RTI), sponsored by China Medical University, Taiwan. Completed at 1 site in Taiwan. Open to participants aged 65 Years to 99 Years, including healthy volunteers. Per ClinicalTrials.gov, last updated 2026-10-21.
Sponsored by China Medical University, Taiwan · Not applicable, Interventional, and Supportive care
Background Inadequate oral health can further compromise general health, particularly in elderly patients who are more susceptible to tooth loss, periodontal disease, and oral infections. These conditions can impair nutrition and overall well-being and are associated with increased risks of systemic diseases, underscoring the critical need for effective oral care in this population. Essential oils and various plant extracts have attracted considerable attention as potential natural medicinal products due to their proven antibacterial, antifungal, antiviral, and antioxidant properties. However, the precise effects and underlying mechanisms through which essential oil mouthwashes promote oral health remain incompletely understood.
Objective This study aims to evaluate the potential of a mouthwash formulated with a blend of tea tree, lemon, and bay laurel essential oils, which have long been recognized for their medicinal properties in alleviating inflammatory signs of intraoral mucosa and promoting both oral and gingival health, in comparison with/ without alcohol-based mouthwashes.
Methods
One hundred five community-dwelling elderly individuals were randomly assigned to one of three groups and used a designated mouthwash for oral care over 21 days. The study groups were as follows:
Expected Results The treatment group of community-dwelling elderly individuals using the essential oil mouthwash is expected to show a significant reduction in oral bacterial load, improvement in gingival inflammation and periodontal symptoms, and a decreased risk of upper respiratory infections compared to the control groups using alcohol-free mouthwashes.
Background The application of mouthwash that decreases viral load in the oral cavity has been reported to be effective in mitigating disease severity, as evidenced by reductions in the length of hospital stays, the need for intensive care admission, and mortality rates (da Silva Santos et al., 2021). While bacteria naturally inhabit the oral cavity, certain bacterial species are responsible for the development of caries and periodontal disease, particularly prevalent among the elderly in the community (Filipovic et al., 2020). Available antimicrobial mouthwashes can either eliminate pathogenic bacteria (bactericidal) or inhibit their growth (bacteriostatic). Additionally, some formulations prevent the development of biofilms, which play a crucial role in the formation of plaque and the progression of dental caries, gingivitis, and periodontal disease (Brookes et al., 2023). The anti-plaque activity of mouthwashes is partly attributed to their antimicrobial properties, with key antiseptic ingredients including chlorhexidine (CHX) and essential oils (Liu et al., 2023). Specifically, chlorhexidine or essential oils formulations can penetrate bacterial biofilms and exert potent antibacterial effects (Ramberg et al., 2016). Additionally, antiseptic mouthwashes complement daily brushing in managing dental plaque and gingivitis (Navabi et al., 2024).
A wide range of chemical and natural antiseptic mouthwashes is readily available over-the-counter (OTC) for managing bad breath (halitosis), dental decay, and gum disease. These products are often used daily without requiring a prescription or guidance from a dental professional. Common active ingredients include chlorhexidine (CHX), hydrogen peroxide (H2O2), cetylpyridinium chloride (CPC), povidone-iodine (PVP-I), and essential oils (EO) (Brookes et al., 2023). Chlorhexidine (CHX) has demonstrated antiviral properties in vitro against specific viruses, including herpes simplex type-1, influenza A, human coronavirus, and severe acute respiratory syndrome-related coronavirus. It appears to be more effective against enveloped viruses than non-enveloped ones (Brookes et al., 2023). Substantial evidence supports the antibacterial effects of oral rinses containing chlorhexidine (CHX) gluconate, significantly reducing bacterial populations on buccal epithelia and pellicles. These rinses are effective against various microorganisms, including Actinomyces naeslundii, Veillonella dispar, Prevotella nigrescens, and Streptococcus species (Babu \& Garcia-Godoy, 2014; Filipovic et al., 2020). Systematic reviews provide robust evidence indicating that commonly used mouthwashes effectively reduce plaque and gingivitis, with chlorhexidine (CHX) widely regarded as the most effective option (Brookes et al., 2023). Beyond its dental applications, CHX mouthwash has also been suggested to lower the risk of ventilator-associated pneumonia in critically ill patients (Hua et al., 2016). The effects of CHX mouthwash on bacterial species have been investigated through studies using in vitro oral microbial communities. For example, when a community of ten oral bacterial species was exposed to CHX, there were notable reductions in Prevotella sp. and Selenomonas infelix. In contrast, other species exhibited less pronounced reductions (McBain et al., 2003). The impact of Listerine® (LIS) on bacterial species has also been assessed using culture-based methods to quantify bacterial levels. One study reported significant reductions in Veillonella sp., Capnocytophaga sp., and Fusobacterium nucleatum in subgingival plaque after two weeks of gargling with LIS (Fine et al., 2007). In another in vitro study involving 40 oral bacterial species, CHX mouthwash was more effective than LIS in reducing all the species tested (Haffajee et al., 2008).
A systematic review revealed that alcohol-free povidone-iodine (PVP-I) mouthwash exhibits significant virucidal activity against SARS-CoV-2 in saliva at concentrations of 0.5% to 1%. However, a 2% concentration did not significantly reduce salivary viral load in one of the included studies. In clinical settings, a 30- to 60-second rinse with 0.5% or 1% PVP-I mouthwash before procedures may effectively reduce the risk of cross-infection, particularly during aerosol-generating procedures (AGPs) involving confirmed, suspected, or asymptomatic COVID-19 patients (Garcia-Sanchez et al., 2022). Povidone-iodine is commonly incorporated into mouthwashes at concentrations of 0.5% to 1.0% and is frequently recommended as a gargle for sore throats due to its potent antiseptic properties. It has demonstrated bactericidal, fungicidal, and virucidal efficacy and is extensively utilized for disinfecting skin and mucous membranes, such as in Betadine products. Beyond oral care, povidone-iodine is used in hand washes, surgical scrubs, and skin disinfectants at concentrations of 7.5% to 10% (Brookes et al., 2023; Stewart, 2022). Its broad-spectrum antimicrobial activity extends to oral microorganisms, including periodontal pathogens, fungi, mycobacteria, viruses, and protozoa, without exhibiting cytotoxic effects on human cells (Schreier et al., 1997). For decades, povidone-iodine mouthwash has been recommended for periodontal site decontamination before invasive procedures to reduce the risk of bacteremia (Brookes et al., 2023).
Essential oils (EOs) are natural extracts derived from various plants. They are recognized for their antibacterial, antifungal, and antioxidant properties, combined with relatively low toxicity (Brookes et al., 2023). Historically, essential oils have been used to enhance physical health, improve appearance, and protect the body from environmental damage. These versatile products serve multiple purposes, such as cleansing, perfuming, altering appearance, correcting body odors, and preserving or maintaining treated areas in optimal condition (Carvalho et al., 2016). Essential oils and their compounds are commonly
optimal condition (Carvalho et al., 2016). Essential oils and their compounds are commonly incorporated into mouthwashes, primarily for their fragrance. However, evidence supporting their antibacterial efficacy against oral microbiota or their use as preservatives remains limited. For instance, benzyl alcohol is used mainly for fragrance and preservation, while its derivative, 2,4-dichlorobenzyl alcohol, is a mild antiseptic for oral and throat applications. Compounds such as limonene, linalool, citral, and cinnamaldehyde have demonstrated significant activity against oral pathogens, with geraniol and anethole showing some antimicrobial potential (Radzki et al., 2022). This systematic review presents the first comprehensive analysis of human trials assessing tea tree oil's therapeutic efficacy and safety. Tea tree oil's therapeutic applications span various medical fields, including dentistry, dermatology, infectious diseases, ophthalmology, and podiatry (Kairey et al., 2023). Listerine® (LIS), a widely used mouthwash containing four plant-derived essential oils-eucalyptol, menthol, methyl salicylate, and thymol-has been proven effective in reducing plaque and gingivitis (Alshehri, 2018). In this study, the selection criteria for essential oils included in mouthwash formulations were based on three key factors: 1) commercial availability, 2) antimicrobial efficacy against harmful anaerobic oral microorganisms, and 3) minimal toxicity to gingival cells.
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This study's enrollment of 150 is above the median of 59 across 286 interventional studies indexed under Dental Plaque.
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Exclusion Criteria:
Participants rinse with 30 mL of an alcohol-free mouthwash containing a blend of tea tree, lemon, and bay laurel essential oils for at least 30 seconds twice daily (morning and evening) after toothbrushing for 21 days.
Other: Essential oil mouthwash
Community-dwelling older adults using an alcohol-free chlorhexidine mouthwash twice daily for 21 days.
Other: Chlorhexidine gluconate alcohol-free mouthwash
Community-dwelling older adults using an alcohol-free povidone-iodine mouthwash twice daily for 21 days.
Other: Povidone-iodine alcohol-free mouthwash
Participants rinse with 30 mL of an alcohol-free mouthwash containing a blend of tea tree, lemon, and bay laurel essential oils for at least 30 seconds twice daily (morning and evening) after toothbrushing for 21 days.
Participants rinse with 30 mL of an alcohol-free chlorhexidine gluconate 2.0 mg/mL mouthwash for at least 30 seconds twice daily (morning and evening) after toothbrushing for 21 days.
Participants rinse with 30 mL of an alcohol-free povidone-iodine mouthwash (available iodine 1 mg/mL) for at least 30 seconds twice daily (morning and evening) after toothbrushing for 21 days.
Change in Oral Health Assessment Tool (OHAT) Score
Overall oral health status assessed by a trained assessor blinded to group allocation using the Oral Health Assessment Tool, which comprises eight domains (lips, tongue, gums and oral tissues, saliva, natural teeth, dentures, oral cleanliness, and dental pain), each scored 0-2. Total score ranges from 0 to 16; higher scores indicate poorer oral health.
Time frame: Baseline (Day 1) and Day 21
Change in Plaque Index Score
Dental plaque accumulation assessed using a plaque-disclosing agent and scored by a trained assessor blinded to group allocation according to the Silness and Löe criteria: 0 = no plaque, 1 = thin film of plaque along the gingival margin, 2 = visible plaque accumulation, 3 = abundant plaque accumulation. Higher scores indicate greater plaque accumulation.
Time frame: Baseline (Day 1) and Day 21
Change in Halitosis Consequences Inventory (ICH) Score
Halitosis-related quality of life assessed using the 18-item self-report Halitosis Consequences Inventory, with each item rated dichotomously for the presence (1) or absence (0) of the corresponding behavioral, emotional, or social consequence of bad breath. Total score ranges from 0 to 18; higher scores indicate greater perceived impact of halitosis.
Time frame: Baseline (Day 1) and Day 21
Change in Stomatitis Index Score
Oral mucosal inflammation assessed by non-invasive visual inspection by a trained assessor blinded to group allocation, using Newton's classification: 0 = no lesions, 1 = localized erythema or pinpoint hyperemia, 2 = diffuse erythema or generalized inflammation, 3 = granular or hyperplastic surface consistent with inflammatory papillary hyperplasia. Higher scores indicate more severe inflammation.
Time frame: Baseline (Day 1) and Day 21
Change in Self-Perceived Oral Health Status
Self-perceived oral health assessed using a single item asking participants to rate their current oral health as good (0), average (1), bad (2), or very bad (3). Higher scores indicate poorer self-perceived oral health.
Time frame: Baseline (Day 1) and Day 21
Change in Geriatric Oral Health Assessment Index (GOHAI) Score
Oral health-related quality of life assessed using the 12-item self-report GOHAI, covering physical function, psychosocial function, and pain or discomfort related to oral health. Each item is rated on a five-point frequency scale (0-4), and the total score is the sum of the 12 items (range 0-48).
Time frame: Baseline (Day 1) and Day 21
Incidence of Upper Respiratory Tract Symptoms
Presence of self-reported upper respiratory tract symptoms (nasal congestion, rhinorrhea, sneezing, or cough), recorded as present or absent and summarized as the number and percentage of participants with any symptom in each group.
Time frame: Baseline (Day 1) and Day 21
Change in Salivary Oral Microbiota Composition
Unstimulated saliva samples collected to characterize oral microbial composition and diversity using \[sequencing method, e.g., 16S rRNA gene sequencing\], including relative abundance of bacterial taxa and alpha- and beta-diversity indices.
Time frame: Baseline (Day 1) and Day 21
Change in Salivary Inflammatory Markers
Concentrations of salivary inflammatory markers \[specify markers, e.g., IL-1β, IL-6, TNF-α\] measured in saliva samples using \[assay method, e.g., enzyme-linked immunosorbent assay\].
Time frame: Baseline (Day 1) and Day 21
Plan to share: No — Individual participant data (IPD) will not be made publicly available because the dataset contains health information from community-dwelling older adults that could compromise participant privacy and confidentiality, and the informed consent obtained did not include provision for public data sharing. De-identified data supporting the published findings may be made available from the corresponding author upon reasonable request, subject to approval by the Institutional Review Board of An Nan Hospital, China Medical University, and in compliance with applicable data protection regulations and institutional data governance policies.
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China Medical University, Taiwan