Clear Aligner Cleaning: Brushing, Chlorhexidine, and BlueM Effects on Oral Microbial Ecology and Periodontal Indices

NCT07307716 | Not Yet Recruiting

• 1 other identifier: REU-ORTHO-ECOALIGN-XO-01
• Study Type: interventional
• Target: 21
• Locations: 1 country
• Sites: 1

Brief Summary

This randomized crossover study will assess how three aligner-cleaning methods affect early supragingival oral microbial ecology and periodontal health in adults using clear aligners. Each participant will complete three (1-month) periods in a randomized sequence with (1-week) washouts between periods:

1. 1.Mechanical toothbrushing with fluoride toothpaste (control),
2. 2.Chlorhexidine (CHX) mouthwash, and
3. 3.an oxygen-based cleanser (BlueM).

Conditions

Gingivitis; Dental Plaque

Keywords

Clear aligners; Chlorhexidine; BlueM; Oxygen-based cleanser; Mechanical toothbrushing; Supragingival plaque; Aligner biofilm; Fusobacterium nucleatum; Prevotella intermedia; MMP-8; Plaque index; Gingival index; Bleeding on probing; Randomized crossover

Outcome Measures

Primary Outcomes (2)

• Fusobacterium nucleatum CFU in supragingival plaque at the gingival margin (index teeth)

  Anaerobic culture enumeration (CFU) of Fusobacterium nucleatum from supragingival plaque collected along the gingival margin of upper/lower first molars and central incisors using the standardized swab/microbrush protocol. Species confirmation per SOP. Culture processing blinded under coded labels (P1-P3) by FA. Metric: CFU per specimen (report also as log10 CFU if analyzed).

  Baseline and end of each 1-month period (end of Period 1, Period 2, Period 3); 1-week washouts between periods.

• Fusobacterium nucleatum CFU on retrieved aligner inner-surface biofilm

  Anaerobic culture enumeration (CFU) of Fusobacterium nucleatum from a standardized inner-surface swab of the used aligner (zones corresponding to gingival margins of index teeth). Species confirmation per laboratory SOP. Culture processing blinded under coded labels (P1-P3) by a single trained operator (FA). Metric: CFU per specimen (report also as log10 CFU if analyzed).

  Baseline and end of each 1-month period (end of Period 1, Period 2, Period 3); 1-week washouts between periods.

Secondary Outcomes (6)

• Prevotella intermedia CFU on retrieved aligner inner-surface biofilm

  Baseline and end of each 1-month period (end of Period 1, Period 2, Period 3); 1-week washouts between periods.

• Prevotella intermedia CFU in supragingival plaque at the gingival margin (index teeth)

  Baseline and end of each 1-month period (end of Period 1, Period 2, Period 3); 1-week washouts between periods.

• MMP-8 concentration/activity in matrix-paired plaque and aligner extracts

  Baseline and end of each 1-month period (end of Period 1, Period 2, Period 3); 1-week washouts between periods.

• Plaque Index (PI) at index teeth

  Baseline and end of each 1-month period (end of Period 1, Period 2, Period 3).

• Gingival Index (GI) at index teeth

  Baseline and end of each 1-month period (end of Period 1, Period 2, Period 3).

Study Arms (3)

Sequence ABC: Brushing then CHX Mouthwash then BlueM

EXPERIMENTAL

Participant completes three 1-month periods in order (A to B to C), each separated by a 1-week washout. A = Mechanical toothbrushing with fluoride toothpaste (control); B = Chlorhexidine (CHX) mouthwash; C = Oxygen-based cleanser (BlueM). Trays are changed every 10 days; aligners are worn 22 h/day; water-only while aligners are in.

Behavioral: Mechanical toothbrushing (fluoride toothpaste); Drug: Chlorhexidine mouthwash (0.2%); Other: Oxygen-based cleanser

Sequence BCA: CHX Mouthwash then BlueM then Brushing

EXPERIMENTAL

Participant completes three 1-month periods in order (B to C to A), each separated by a 1-week washout. A = Mechanical toothbrushing with fluoride toothpaste (control); B = CHX mouthwash; C = Oxygen-based cleanser (BlueM). Trays are changed every 10 days; aligners are worn 22 h/day; water-only while aligners are in.

Behavioral: Mechanical toothbrushing (fluoride toothpaste); Drug: Chlorhexidine mouthwash (0.2%); Other: Oxygen-based cleanser

Sequence CAB: BlueM then Brushing then CHX Mouthwash

EXPERIMENTAL

Participant completes three 1-month periods in order (C to A to B), each separated by a 1-week washout. A = Mechanical toothbrushing with fluoride toothpaste (control); B = CHX mouthwash; C = Oxygen-based cleanser (BlueM). Trays are changed every 10 days; aligners are worn 22 h/day; water-only while aligners are in.

Behavioral: Mechanical toothbrushing (fluoride toothpaste); Drug: Chlorhexidine mouthwash (0.2%); Other: Oxygen-based cleanser

Interventions

Mechanical toothbrushing (fluoride toothpaste) BEHAVIORAL

Control oral-hygiene period using mechanical toothbrushing with a fluoride-containing toothpaste, per standardized study instructions, for the assigned 1-month period. No chlorhexidine or oxygen-based cleanser is used during this period; aligners are removed for oral-hygiene procedures per protocol.

Also known as: Colgate Total

Sequence ABC: Brushing then CHX Mouthwash then BlueM; Sequence BCA: CHX Mouthwash then BlueM then Brushing; Sequence CAB: BlueM then Brushing then CHX Mouthwash

Chlorhexidine mouthwash (0.2%) DRUG

Chlorhexidine mouthwash used per standardized study instructions for the assigned 1-month period. No oxygen-based cleanser is used during this period; aligners are removed for mouthrinse use and oral-hygiene procedures per protocol. A 1-week washout without cleaning products separates periods.

Also known as: CHX

Sequence ABC: Brushing then CHX Mouthwash then BlueM; Sequence BCA: CHX Mouthwash then BlueM then Brushing; Sequence CAB: BlueM then Brushing then CHX Mouthwash

Oxygen-based cleanser OTHER

Oxygen-based cleanser (BlueM) used per standardized study instructions for the assigned 1-month period. No chlorhexidine is used during this period; aligners are removed for product use and oral-hygiene procedures per protocol. A 1-week washout without cleaning products separates periods.

Also known as: BlueM

Sequence ABC: Brushing then CHX Mouthwash then BlueM; Sequence BCA: CHX Mouthwash then BlueM then Brushing; Sequence CAB: BlueM then Brushing then CHX Mouthwash

Eligibility Criteria

• Age: 18 Years+
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64), Older Adult (65+)

You may qualify if:

• Adults (≥18 years).
• Undergoing or scheduled for treatment with clear aligners (CA).
• Presence of upper and lower first molars and central incisors.
• Periodontal health at baseline (no sites with PD ≥ 4 mm; no clinical attachment loss at index teeth; BI/PI within normal limits).
• No periodontal therapy within the previous 6 months.
• Able and willing to comply with study instructions and follow-up visits; signed informed consent.
• Sufficient remaining CA treatment (clinician-estimated): ≥ 14 weeks to safely cover three 1 month protocol periods plus washouts.
• Minimum planned aligners remaining: ≥ 11 aligners (assuming 10 days/aligner) or equivalent continuous CA duration per treating orthodontist.

You may not qualify if:

• Patients requiring orthognathic surgery
• Uncontrolled systemic diseases (e.g., uncontrolled diabetes mellitus, autoimmune disease).
• Pregnancy or lactation.
• Use of systemic antibiotics and/or anti-inflammatory drugs within the last 3 months.
• Periodontal therapy within the last 6 months.
• Current tobacco use (smoking/smokeless) or vaping.
• Limited/express CA plans (e.g., ≤ 11 aligners or expected completion \< 14 weeks).
• Anticipated early completion of CA or major treatment changes (e.g., switch to fixed appliances) within the next \~4 months.
• \- Extensive caries, defective restorations, or prostheses affecting index teeth/sites.

Sponsors & Collaborators

• Riyadh Elm University: lead

Study Sites (1)

Riyadh Elm University - Orthodontic Clinics

Riyadh, Riyadh Region, Saudi Arabia

Location

Related Publications (3)

• Heller D, Helmerhorst EJ, Gower AC, Siqueira WL, Paster BJ, Oppenheim FG. Microbial Diversity in the Early In Vivo-Formed Dental Biofilm. Appl Environ Microbiol. 2016 Jan 8;82(6):1881-8. doi: 10.1128/AEM.03984-15.

  PMID: 26746720 BACKGROUND

• Zhao R, Huang R, Long H, Li Y, Gao M, Lai W. The dynamics of the oral microbiome and oral health among patients receiving clear aligner orthodontic treatment. Oral Dis. 2020 Mar;26(2):473-483. doi: 10.1111/odi.13175. Epub 2020 Jan 8.

  PMID: 31418980 BACKGROUND

• Miguel YD, Alcântara PR, Toyofuku ACMM, Shimizu RH. Periodontal monitoring in orthodontic treatment with orthodontic aligners: proof of concept. Rev Odontol UNESP. 2024;53:e20240016. doi:10.1590/1807-2577.01624

  BACKGROUND

MeSH Terms

Conditions

Gingivitis; Dental Plaque

Interventions

hydrated silica gel-based toothpaste

Condition Hierarchy (Ancestors)

Infections; Gingival Diseases; Periodontal Diseases; Mouth Diseases; Stomatognathic Diseases; Dental Deposits; Tooth Diseases

Study Officials

• Dr. Faisal A Althobaiti, BDS

  Riyadh Elm University

  PRINCIPAL INVESTIGATOR

• Dr. Nancy M Ajwa, MSc

  Riyadh Elm University

  STUDY DIRECTOR

Central Study Contacts

Faisal A Althobaiti, BDS

CONTACT

+966594496974; faisal.althobaiti2024@student.riyadh.edu.sa

Nancy M Ajwa, MSc

CONTACT

+966554804564; nancy.ajwa@riyadh.edu.sa

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: RANDOMIZED
• Masking: SINGLE
• Who Masked: OUTCOMES ASSESSOR
• Masking Details: Culture enumeration from plaque and retrieved aligners will be processed under coded labels (P1-P3) by a single trained operator (FA) who is masked to protocol assignment. Microscopy and clinical assessments will be conducted unblinded after decoding.
• Purpose: PREVENTION
• Intervention Model: CROSSOVER
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Principal Investigator

Model Details: Three 1-month periods in randomized sequence (ABC/BCA/CAB) with 1-week washouts between the groups. A = mechanical toothbrushing (fluoride toothpaste), B = chlorhexidine mouthwash, C = oxygen-based cleanser (BlueM). Algners changed every 10 days; water-only while aligners are in

Study Record Dates

• First Submitted: December 15, 2025
• First Posted: December 29, 2025
• Study Start: February 1, 2026
• Primary Completion (Estimated): October 1, 2026
• Study Completion (Estimated): October 1, 2026
• Last Updated: January 8, 2026

Record last verified: 2026-01

Locations

SA (1)