Oral-Spray Bacillus Spore Probiotic in Preventing and Supporting Treatment of Dental Caries in Children

NCT07241052 | Not Yet Recruiting DMC

• 1 other identifier: TNLS.2025.S
• Study Type: interventional
• Target: 130
• Locations: 0 countries
• Sites: N/A

Brief Summary

Despite advances in oral health care, cavities remain highly prevalent worldwide, especially among children. Conventional treatment methods focus on removing damaged tooth tissue but fail to completely eliminate or control pathogenic bacteria in the oral cavity, leading to a high risk of caries recurrence in the same or adjacent teeth. Probiotics are a promising biological therapy that can help restore balance in the oral microbiome. Among probiotics, Bacillus species are unique because they form spores that make them stable under harsh conditions, such as heat and storage. They can also produce enzymes and natural compounds with antibacterial, anti-inflammatory, and protective effects. These features make Bacillus probiotics strong candidates for oral health applications. This study is designed to evaluate the safety and efficacy of an oral-spray probiotic containing Bacillus subtilis ANA48 and Bacillus clausii ANA39 (LiveSpo® Smile CARE) in preventing and supporting the treatment of tooth decay in children. We hypothesize that this probiotic spray, when used regularly, will be safe and well-tolerated, while effectively reducing harmful bacteria in the mouth, preventing the progression of cavities, and improving overall oral health. Study Design:

• Sample Size: 130 participants
• Study Location: a primary school in Hanoi, Vietnam Intervention Description: A total of 130 eligible participants will be randomly assigned to two groups (n = 65 per group). Placebo Group will receive a 0.9% NaCl physiological saline, spray twice daily (morning and afternoon). Each application consists of two sprays at three sites (both dental arches and the front teeth), for a total of six sprays per use, continued for three consecutive months. Smile Care Group will receive an oral-spray probiotics containing ≥ 1 billion CFU/mL × 20 mL of Bacillus subtilis and Bacillus clausii (LiveSpo Smile CARE). The dosage, frequency, and duration of use will be the same as in the placebo group. All participants will receive the same standard dental care throughout the study, including the use of identical toothpaste, supervised weekly fluoride rinsing with 0.2% sodium fluoride (NaF), and scheduled follow-up visits. The probiotic and placebo sprays will be packaged in blinded bottles and administered according to the study protocol to maintain objectivity. Study Duration: 12-18 months

Conditions

Dental Caries

Keywords

Dental caries; Children; probiotics; Oral health; Oral-spray; Bacillus spores

Outcome Measures

Primary Outcomes (1)

• Decayed Teeth Index

  Changes will be calculated for each participant based on the following parameters: the number of decayed teeth assessed using the International Caries Detection and Assessment System (ICDAS, codes 0-6), progressive caries rate, early caries rate, and early caries rate (code 1). These parameters will be evaluated at month 3 compared with day 0. Month 1 and Month 2 will be collected as supplementary exploratory timepoints.

  Day 0, and months 1, 2, 3

Secondary Outcomes (2)

• Changes in the concentration of pathogenic bacteria in deep carious dentin and saliva samples

  Day 0 and months 1, 2, 3

• Changes in Inflammatory Cytokines and IgA levels in deep carious dentin and saliva samples

  Day 0 and months 1, 2, 3

Other Outcomes (2)

• Changes in the oral microbiome at month 1 compared to day 0

  Day 0 and 1 month

• Detection of Probiotic Bacillus Spores

  Day 0 and months 1, 2, 3

Study Arms (2)

Placebo

PLACEBO COMPARATOR

Participants allocated to the placebo group will receive a 0.9% NaCl physiological saline spray (20 mL bottle). The spray will be applied directly to carious teeth and adjacent teeth, with a total of six sprays per application, administered twice daily (morning and afternoon), under teacher supervision. All participants will maintain identical daily oral hygiene practices, including the use of fluoride toothpaste and supervised rinsing with 0.2% sodium fluoride mouthwash once per week. The intervention will be continued for a duration of 3 months.

Drug: oral-spray 0.9% NaCl physiological saline solution

Smile Care

EXPERIMENTAL

Children will receive LiveSpo® Smile CARE, an oral-spray probiotic containing ≥ 1 × 10⁹ CFU/ mL Bacillus subtilis and Bacillus clausii in 20 mL of 0.9% sodium chloride solution. The spray will be administered with the same dosage, frequency, timing, and supervision as the placebo group. The intervention will be continued for a duration of 3 months.

Combination Product: Smile Care

Interventions

oral-spray 0.9% NaCl physiological saline solution DRUG

The oral-spray 0.9% NaCl physiological saline is prepared by extracting 20 mL from a 500 mL polypropylene bottle of 0.9% NaCl intravenous infusion (B. Braun, Germany; product declaration No. VD-32732-19). The solution is contained in an opaque plastic spray bottle identical to that used for the Smile Care.

Placebo

Smile Care COMBINATION_PRODUCT

LiveSpo® Smile CARE is manufactured as a Class-A medical device (product declaration No. 250002196/PCBA-HN) in compliance with manufacturing standards approved by the Hanoi Department of Health, Ministry of Health. The product is formulated as a 0.9% NaCl physiological saline solution plus Bacillus subtilis ANA48 and Bacillus clausii ANA39 at total concentration of ≥ 1 × 10⁹ CFU/mL (20 billion CFU/20 mL suspension).

Smile Care

Eligibility Criteria

• Age: 7 Years - 10 Years
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Child (0-17)

You may qualify if:

• Children (male or female) aged 7-10 years, currently enrolled in primary school (grades 2-4).
• At least one decayed tooth (any stage) at the time of screening.
• Commitment to comply with study requirements, including refraining from using any other probiotic products for the ear, nose, and throat that could affect study outcomes.
• Written informed consent obtained from a parent or legal guardian.

You may not qualify if:

• Presence of severe chronic disease, allergy, or use of antibiotics within 1 month prior to study initiation.
• Use of oral or throat probiotic sprays/lozenges within the past 30 days (excluding gastrointestinal probiotics).
• Participation in another clinical study within the past 30 days.
• Acute oral conditions (e.g., abscess, severe gingivitis) interfering with examination or intervention.
• Currently receiving dental treatment at specialized dental/oral health facilities.
• History of congenital immunodeficiency or infectious disease.
• Psychiatric disorders or sleep disturbances.
• Known sensitivity or allergy to components of the investigational product.
• Hard tissue lesions not caused by dental caries.
• Supernumerary teeth or teeth not part of the functional dental arch.
• Carious teeth with pain lasting \>10 seconds during examination.
• Carious teeth with visible pulp hyperemia (high risk of pulp complications).
• Teeth showing signs of pulp disease, pulp necrosis, or periapical pathology.
• Teeth previously treated with endodontic therapy or permanently restored with crowns.
• Primary teeth with mobility grade ≥2 (due to disease or natural exfoliation).

Sponsors & Collaborators

• Anabio R&D: lead
• Hanoi Medical University: collaborator

Related Publications (9)

• Kim H, Han CY, Eun SH, Kim MG, Im AR, Lee B. Inhibitory effects of Bacillus velezensis ID-A01 supernatant against Streptococcus mutans. BMC Microbiol. 2023 Nov 24;23(1):362. doi: 10.1186/s12866-023-03114-2.

  PMID: 37996837 RESULT

• Jindal G, Pandey RK, Agarwal J, Singh M. A comparative evaluation of probiotics on salivary mutans streptococci counts in Indian children. Eur Arch Paediatr Dent. 2011 Aug;12(4):211-5. doi: 10.1007/BF03262809.

  PMID: 21806906 RESULT

• Aleti G, Sessitsch A, Brader G. Genome mining: Prediction of lipopeptides and polyketides from Bacillus and related Firmicutes. Comput Struct Biotechnol J. 2015 Mar 24;13:192-203. doi: 10.1016/j.csbj.2015.03.003. eCollection 2015.

  PMID: 25893081 RESULT

• Weisser K, Ganz R, Olafsson A, Wyler F, Rutishauser M. Bronchopulmonary dysplasia in the premature baby. Respiration. 1970;27:Suppl:36-40. No abstract available.

  PMID: 4929038 RESULT

• Bustamante M, Oomah BD, Mosi-Roa Y, Rubilar M, Burgos-Diaz C. Probiotics as an Adjunct Therapy for the Treatment of Halitosis, Dental Caries and Periodontitis. Probiotics Antimicrob Proteins. 2020 Jun;12(2):325-334. doi: 10.1007/s12602-019-9521-4.

  PMID: 30729452 RESULT

• Simon-Soro A, Mira A. Solving the etiology of dental caries. Trends Microbiol. 2015 Feb;23(2):76-82. doi: 10.1016/j.tim.2014.10.010. Epub 2014 Nov 27.

  PMID: 25435135 RESULT

• Lu Y, Lin Y, Li M, He J. Roles of Streptococcus mutans-Candida albicans interaction in early childhood caries: a literature review. Front Cell Infect Microbiol. 2023 May 16;13:1151532. doi: 10.3389/fcimb.2023.1151532. eCollection 2023.

  PMID: 37260705 RESULT

• Takahashi N, Nyvad B. The role of bacteria in the caries process: ecological perspectives. J Dent Res. 2011 Mar;90(3):294-303. doi: 10.1177/0022034510379602. Epub 2010 Oct 5.

  PMID: 20924061 RESULT

• Sampaio-Maia B, Caldas IM, Pereira ML, Perez-Mongiovi D, Araujo R. The Oral Microbiome in Health and Its Implication in Oral and Systemic Diseases. Adv Appl Microbiol. 2016;97:171-210. doi: 10.1016/bs.aambs.2016.08.002. Epub 2016 Sep 21.

  PMID: 27926431 RESULT

MeSH Terms

Conditions

Dental Caries

Interventions

Oral Sprays

Condition Hierarchy (Ancestors)

Tooth Demineralization; Tooth Diseases; Stomatognathic Diseases

Intervention Hierarchy (Ancestors)

Aerosols; Colloids; Complex Mixtures; Dosage Forms; Pharmaceutical Preparations

Study Officials

• Nga TT Pham, PhD. MD

  Hanoi Medical University

  STUDY CHAIR

• Thu TH Nguyen, Assoc. Prof.

  Hanoi Medical University

  PRINCIPAL INVESTIGATOR

• Anh TV Nguyen, Assoc. Prof.

  Spobio Research Center, Anabio R&D

  PRINCIPAL INVESTIGATOR

Central Study Contacts

Nga TT Pham, PhD. MD

CONTACT

(84) 0983042975; Tuyetnga@hmu.edu.vn

Thu TH Nguyen, Assoc. Prof.

CONTACT

(84) 0903211266; nguyenhoaithu@hmu.edu.vn

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: RANDOMIZED
• Masking: TRIPLE
• Who Masked: PARTICIPANT, CARE PROVIDER, INVESTIGATOR
• Masking Details: LiveSpo Smile CARE and placebo are indistinguishable regarding taste and smell. The color and turbidity of LiveSpo Smile CARE suspension are unrecognizable to investigators, trainers, caregivers, and paticipants due to opaque plastic container. Only the independent statistician is aware of the group codes.
• Purpose: PREVENTION
• Intervention Model: PARALLEL
• Sponsor Type: INDUSTRY
• Responsible Party: SPONSOR

Model Details: Blinded, randomized, and controlled clinical trial

Study Record Dates

• First Submitted: November 17, 2025
• First Posted: November 21, 2025
• Study Start: December 1, 2025
• Primary Completion (Estimated): October 30, 2026
• Study Completion (Estimated): May 1, 2027
• Last Updated: November 21, 2025

Record last verified: 2025-11

Data Sharing

• IPD Sharing: Will share
• Shared Documents: STUDY PROTOCOL, SAP, ICF, CSR
• Time Frame: Data requests can be submitted starting 9 months after article publication and the data will be made accessible for up to 24 months. Extensions will be considered on a case-by-case basis.
• Access Criteria: Access to trial IPD can be requested by qualified researchers engaging in independent scientific research and will be provided following review and approval of a study protocol, informed consent form (ICF), and clinical study reports (CSR). For more information or to submit a request, please contact anabio.rd2021@gmail.com