Validate the Effect of Calcium Oral Spray on Early Childhood Caries

Validate the Effect of T-spray on ECC in Clinical Trials

This exploratory randomized clinical trial aimed to evaluate the effects of calcium-containing oral sprays on the primary clinical outcome, fluorescence-detectable plaque-associated area measured by quantitative light-induced fluorescence (QLF). Fifteen children were randomly assigned to placebo, Formula 1 (0.3% calcium lactate), or Formula 2 (0.3% calcium lactate plus 225 ppm fluoride) oral sprays for two months. Secondary outcomes included salivary calcium concentration and exploratory plaque microbiome analysis.

Study Overview

• Status: Completed
• Conditions: Early Childhood Caries (ECC); Dental Caries Tooth Demineralization

Detailed Description

Calcium-containing biomimetic materials have been investigated as adjunctive approaches for managing early enamel demineralization. This exploratory randomized clinical trial evaluated the effects of calcium-containing oral sprays on fluorescence-detectable plaque-associated area measured by quantitative light-induced fluorescence (QLF). Fifteen children were randomly assigned to placebo, Formula 1 (0.3% calcium lactate), or Formula 2 (0.3% calcium lactate plus 225 ppm fluoride) oral sprays for two months. The primary clinical outcome was fluorescence-detectable plaque-associated area measured by QLF. Secondary outcomes included salivary calcium concentration measured using the arsenazo III colorimetric assay, and exploratory plaque microbiome analysis using 16S rRNA gene sequencing.

• Study Type: Observational
• Enrollment (Actual): 15

Contacts and Locations

Study Locations

• Taiwan
  • Taiwan
    • Taipei, Taiwan, Taiwan, 11490
      • Tri-Service General Hospital

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Child
• Accepts Healthy Volunteers: No

• Sampling Method: Probability Sample

Study Population

A randomized clinical trial (RCT) will be conducted in a double-blind manner with 15 children presenting with early carious lesions in Tri-Service General Hospital, Taipei, Taiwan. Participants will be randomly assigned to one of three groups: placebo (control) and two formulations of calcium spray.

Description

Inclusion Criteria:

Required participants to have an ICDAS code ≥ 1 and ≤ 3 as determined by a clinician.

Exclusion Criteria:

1. Patient has received caries removal and restoration.
2. Patient has received fluoridation within 3 months.
3. The parents do not agree to participate in the study.
4. Patient has ICDAS code 4 - code 6

Study Plan

How is the study designed?

Number of groups / cohorts

3

Cohorts and Interventions

Group / Cohort
placebo oral spray; formula 1 (0.3% calcium) oral spray
formula 2 (0.3% calcium plus 225 ppm fluoride) oral spray
formula 1 (0.3% calcium); Patient receiving oral spays containing 0.3% calcium solution for more than 2 month.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
salivary calcium	Saliva samples collected during the clinical trial will be centrifuged at 3000 rpm for 10 minutes to obtain the clarified supernatant, which will then be diluted tenfold for calcium measurement. Calcium concentration will be determined colorimetrically using arsenazo III in a microplate reader. The calcium-arsenazo III complex will be measured spectrophotometrically at 652 nm, with absorbance proportional to calcium concentration in the sample.	Saliva samples will be collected at the time of recruitment and 2 months after the application of the clinical trial.

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Plaque bacterial composition	Three out of 15 participants will be selected for bacterial sampling (pre- and post-treatment). Plaque samples will be collected using a periodontal probe and stored in 1.5 mL tubes containing 500 μL of preservation solution at -80°C. Bacterial DNA will be extracted following the manufacturer's instructions using the EasyPure Bacteria Genomic DNA Kit (TransGen Biotech, Cat No. EE161). DNA purity and concentration were verified using a Nanodrop spectrophotometer. The V3 and V4 regions of the 16S rRNA gene will be amplified with primers (CCTACGGRRBGCASCAGKVRVGAAT and GGACTACNVGGGTWTCTAATCC) and the MetaVX Library Preparation Kit. Taxonomic classification will be performed using the Silva 138 reference database and the Ribosomal Database Project (RDP).	Plaque samples will be collected at the time of recruitment and 2 months after the application of the clinical trial.

Other Outcome Measures

Outcome Measure	Measure Description	Time Frame
cariogenicity area	In the clinical trial, QLF imaging produced three outputs: (1) an original photograph, (2) a binary image highlighting carious lesions, and (3) a fluorescent image. The QLF-area (%) and its change (ΔQLF-area) were measured and compared.	QLF images of maxillary front teeth will be collected at the time of recruitment and 2 months after the clinical trial.

Collaborators and Investigators

• Sponsor: Tri-Service General Hospital (TSGH)

Study record dates

Study Major Dates

• Study Start (Actual): June 11, 2023
• Primary Completion (Actual): August 5, 2024
• Study Completion (Actual): November 11, 2025

Study Registration Dates

• First Submitted: November 25, 2025
• First Submitted That Met QC Criteria: November 25, 2025
• First Posted (Actual): December 8, 2025

Study Record Updates

• Last Update Posted (Actual): July 14, 2026
• Last Update Submitted That Met QC Criteria: July 12, 2026
• Last Verified: July 1, 2026

More Information

Terms related to this study

• Keywords: ECC; Oral spray
• Other Study ID Numbers: A202305084; 801GB112200 (Other Grant/Funding Number: Tri-Service General Hospital, Tapiei, Taiwan)

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO
• IPD Plan Description: Patients who will be recruited are children under 15 years old, whose privacy should be further protected.

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No