Oral-Spray Bacillus Spore Probiotics for Periodontal Disease Prevention and Supprotive Treatment in the Elderly

NCT07258303 | Recruiting DMC

• 1 other identifier: TNLS.2025.NC
• Study Type: interventional
• Target: 90
• Locations: 1 country
• Sites: 1

Brief Summary

Periodontal diseases are strongly associated with dysbiosis of the oral microbiome within the biofilm. Key pathogens, including Streptococcus mutans, Aggregatibacter actinomycetemcomitans, and Porphyromonas gingivalis, contribute to caries progression and periodontal inflammation. Given the limitations of antibiotic use and their potential side effects, probiotics represent a promising biological approach to restore microbial balance and support oral health. In this study, researchers propose that direct application of probiotic spray into the oral cavity is safe and effective in alleviating typical symptoms of periodontal disease, particularly by reducing pathogenic bacterial density. The objective of this study is to evaluate the safety and efficacy of a oral-spray Bacillus spore probiotics (LiveSpo Smile CARE), containing Bacillus subtilis ANA48 and Bacillus clausii ANA39 at a concentration of ≥ 1 billion CFU/mL x 20 mL formulation, for the prevention and supportive treatment of periodontal disease. Study Design:

• Sample Size: 90 participants
• Study Location: an elderly care center in Hanoi, Vietnam Intervention Description: A total of 90 eligible participants will be randomly assigned to two groups (n = 45 per group).
• The Placebo Group will receive 0.9% NaCl physiological saline, spray twice daily (afternoon and evening), with two sprays per application at three sites (both dental arches and the front teeth), totaling six sprays per use, for four consecutive weeks.
• The 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), administered with the same dosage and frequency as the placebo group. All participants will receive blinded, coded spray bottles to ensure objectivity. Both groups will be instructed to use the same standardized toothpaste during the study period. Caregivers will monitor the health status of participants and record relevant information in their medical records. During the study, caregivers will be instructed not to provide any additional probiotic oral care products or the solution contains similar components. Study Duration: 12-18 months.

Conditions

Periodontal Disease; Oral Deseases

Keywords

Periodontal Disease; The elderly; pathogenic bacterial densities; Bacillus; Spores; oral-spray probiotics

Outcome Measures

Primary Outcomes (2)

• Oral health indicators

  Percentage of teeth classified according to a grading scale (good, moderate, and severe) for the following parameters: Simple Debris Index (DI-S), Gingival Index (GI), Bleeding on Probing (BOP), and Halitosis (score 1-4) is assessed at various time points relative to day 0.

  Days 0, 7, 14, 28 and 42

• Periodontal parameters

  Periodontal parameters are assessed for the following: Recession of the Gingival Margin (REC, mm), Probing Depth (PD, mm), and Clinical Attachment Loss (CAL, mm). These parameters are assessed at various time points relative to day 0.

  Days 0, 7, 14, 28 and 42

Secondary Outcomes (2)

• Changes in pro-inflammatory cytokine and IgA levels in gingival crevicular fluid samples at days 0, 7, 14, 28, and 42

  Days 0, 7, 14, 28, and 42

• Changes in the concentration of pathogenic bacteria around the teeth

  Days 0, 7, 14, 28, and 42

Other Outcomes (1)

• Changes in the oral microbiome at day 28 compared to day 0

  Days 0 and 28

Study Arms (2)

Placebo

PLACEBO COMPARATOR

Participants allocated to the placebo group will receive a 0.9% NaCl physiological saline spray. The solution will be self-administered via spray, twice daily-once in the afternoon and once at night. At each administration, participants will apply two sprays to each side of the dental arch and two sprays to the incisor region, resulting in a total of six sprays per application. This regimen will be maintained consistently for a period of 4 weeks.

Drug: oral-spray 0.9% NaCl physiological saline solution

Smile Care

EXPERIMENTAL

Participants assigned to the experimental group will receive LiveSpo® Smile CARE (20 mL), formulated as an oral-spray probiotics containing ≥ 1 × 10⁹ CFU/ mL of Bacillus subtilis and Bacillus clausii, of 0.9% sodium chloride solution. The suspension will be self-administered via oral spray twice daily once in the afternoon and once at night. At each administration, participants will apply two sprays to each side of the dental arch and two sprays to the incisor region, totaling six sprays per application. The intervention will be continued for a duration of 4 weeks.

Combination Product: LiveSpo Smile CARE

Interventions

LiveSpo Smile CARE COMBINATION_PRODUCT

LiveSpo Smile CARE is manufactured as a Class-A medical device (product declaration 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 billion CFU/ (20 billion CFU/20 mL suspension)

Smile Care

oral-spray 0.9% NaCl physiological saline solution DRUG

The oral-spray 0.9% NaCl physiological saline solution 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 administration of Smile Care.

Also known as: Registration number: VD-32723-19

Placebo

Eligibility Criteria

• Age: 35 Years+
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64), Older Adult (65+)

You may qualify if:

• Patients aged 35 years and older.
• Patients diagnosed with periodontal disease based on clinical examination, including gingivitis and periodontitis.
• Patients presenting with at least 10 natural teeth in the dental arch.
• Patients with stable general health status and able to comply with study procedures.
• Patients (or their legal guardians) who provide written informed consent and voluntarily agree to participate in the study.

You may not qualify if:

• Patients with unstable general health, acute systemic or oral diseases, or malignant conditions that may interfere with participation.
• Patients who are currently receiving treatment for periodontal disease at specialized dental facilities.
• Patients currently using any therapeutic or adjunctive products for the management of periodontal disease.

Sponsors & Collaborators

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

Study Sites (1)

Tuyet Thai Senior Care Center

Hanoi, Hanoi, 100000, Vietnam

RECRUITING

Related Publications (12)

• Huang Q, Dong X. Prevalence of periodontal disease in middle-aged and elderly patients and its influencing factors. Am J Transl Res. 2022 Aug 15;14(8):5677-5684. eCollection 2022.

  PMID: 36105065 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

• Dang HT, Tran DM, Phung TTB, Bui ATP, Vu YH, Luong MT, Nguyen HM, Trinh HT, Nguyen TT, Nguyen AH, Van Nguyen AT. Promising clinical and immunological efficacy of Bacillus clausii spore probiotics for supportive treatment of persistent diarrhea in children. Sci Rep. 2024 Mar 18;14(1):6422. doi: 10.1038/s41598-024-56627-9.

  PMID: 38494525 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

• Bader J, Albin A, Stahl U. Spore-forming bacteria and their utilisation as probiotics. Benef Microbes. 2012 Mar 1;3(1):67-75. doi: 10.3920/BM2011.0039.

  PMID: 22348911 RESULT

• Graves D. Cytokines that promote periodontal tissue destruction. J Periodontol. 2008 Aug;79(8 Suppl):1585-91. doi: 10.1902/jop.2008.080183.

  PMID: 18673014 RESULT

• Fontana M, Zero DT. Assessing patients' caries risk. J Am Dent Assoc. 2006 Sep;137(9):1231-9. doi: 10.14219/jada.archive.2006.0380.

  PMID: 16946427 RESULT

• Teughels W, Durukan A, Ozcelik O, Pauwels M, Quirynen M, Haytac MC. Clinical and microbiological effects of Lactobacillus reuteri probiotics in the treatment of chronic periodontitis: a randomized placebo-controlled study. J Clin Periodontol. 2013 Nov;40(11):1025-35. doi: 10.1111/jcpe.12155. Epub 2013 Sep 15.

  PMID: 24164569 RESULT

• Amaliya A, Laine ML, Delanghe JR, Loos BG, Van Wijk AJ, Van der Velden U. Java project on periodontal diseases: periodontal bone loss in relation to environmental and systemic conditions. J Clin Periodontol. 2015 Apr;42(4):325-32. doi: 10.1111/jcpe.12381. Epub 2015 Mar 24.

  PMID: 25683157 RESULT

• Hajishengallis G, Lamont RJ. Beyond the red complex and into more complexity: the polymicrobial synergy and dysbiosis (PSD) model of periodontal disease etiology. Mol Oral Microbiol. 2012 Dec;27(6):409-19. doi: 10.1111/j.2041-1014.2012.00663.x. Epub 2012 Sep 3.

  PMID: 23134607 RESULT

• Perez-Chaparro PJ, Duarte PM, Shibli JA, Montenegro S, Lacerda Heluy S, Figueiredo LC, Faveri M, Feres M. The Current Weight of Evidence of the Microbiologic Profile Associated With Peri-Implantitis: A Systematic Review. J Periodontol. 2016 Nov;87(11):1295-1304. doi: 10.1902/jop.2016.160184. Epub 2016 Jul 15.

  PMID: 27420109 RESULT

• Perez-Chaparro PJ, Goncalves C, Figueiredo LC, Faveri M, Lobao E, Tamashiro N, Duarte P, Feres M. Newly identified pathogens associated with periodontitis: a systematic review. J Dent Res. 2014 Sep;93(9):846-58. doi: 10.1177/0022034514542468. Epub 2014 Jul 29.

  PMID: 25074492 RESULT

MeSH Terms

Conditions

Periodontal Diseases

Interventions

Oral Sprays

Condition Hierarchy (Ancestors)

Mouth 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) 903211266; 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: Blind, randomized, and controlled clinical trial

Study Record Dates

• First Submitted: November 18, 2025
• First Posted: December 2, 2025
• Study Start: December 20, 2025
• Primary Completion (Estimated): October 30, 2026
• Study Completion (Estimated): May 1, 2027
• Last Updated: January 21, 2026

Record last verified: 2026-01

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

Locations

VN (1)