Significant elevation of salivary human neutrophil peptides 1-3 levels by probiotic milk in preschool children with severe early childhood caries: a randomized controlled trial.

OBJECTIVES: To determine salivary human neutrophil peptides 1-3 (HNP1-3) levels in caries-free preschool children and in those with early childhood caries (ECC) or severe-ECC, in a daily probiotic group, receiving reconstituted milk with the probiotic Lactobacillus paracasei SD1 once daily; a triweekly probiotic group, receiving the probiotic milk 3 days a week; and a placebo group. MATERIALS AND METHODS: Oral examination and unstimulated whole saliva collection were conducted in 354 children at baseline, 6 months after intervention (T6), and after probiotic discontinuation (T12). Of the 354, adequate volume of saliva samples from 268 children were simultaneously analyzed for Streptococcus mutans and total lactobacilli levels using qPCR and for HNP1-3 levels using ELISA. RESULTS: In the severe-ECC status, significant increases in the median HNP1-3 levels at T12 were found in both daily and triweekly probiotic groups (p < 0.001). The median S. mutans levels in the daily group were significantly decreased at T6 and T12 (p < 0.01), whereas the median total lactobacilli levels were significantly increased at T6 (p < 0.001). Significantly inverse correlations between altered HNP1-3 and S. mutans levels and significant decreases in caries progression were found in both probiotic groups (p < 0.05). CONCLUSIONS: In the severe-ECC status, daily or triweekly consumption of L. paracasei SD1 significantly enhanced salivary HNP1-3 levels, but reduced S. mutans levels, possibly resulting in reduction of caries progression. CLINICAL RELEVANCE: Significant enhancement of salivary HNP1-3 levels by probiotic consumption is associated with reduction in S. mutans levels, consistent with diminished caries progression in children with severe-ECC.

Efficacy of Probiotic Milk for Caries Regression in Preschool Children: A Multicenter Randomized Controlled Trial.

This multicenter double-blind randomized controlled trial aimed to determine the efficacy of daily or triweekly consumption of reconstituted milk powder, containing Lactobacillus paracaseiSD1, in preschool children for caries prevention. A 6-month intervention was conducted in 487 children (aged 37.6 ± 9.2 months) with ≤4 decayed teeth from 8 childcare centers. Using stratified block randomization, participants were randomly allocated into 3 groups, comprising: (1) daily probiotic, receiving probiotic milk once daily, (2) triweekly probiotic, randomly receiving probiotic milk 3 days a week and the placebo milk for the remaining 4 days, and (3) placebo, receiving milk without probiotics. Each tooth surface was assessed for caries status using the modified Nyvad criteria at baseline (T0), 6 (T6) and 12 (T12) months after T0. Study outcomes were caries transition, including caries progression and regression during the T0-T6, T6-T12 and T0-T12 periods. Negative binomial regression with a generalized linear model was used to estimate the caries outcomes, which were reported as incidence rate ratios (IRRs) and 95% confidence intervals (CIs). At baseline, caries prevalence and mean decayed surfaces were 81.9% and 7.29 ± 7.60, respectively. Three groups were balanced at baseline with an overall dropout rate of 25.2%. After adjusting for age, sex, and the number of noncavitated and cavitated caries at baseline, a decreased caries risk during T0-T6 was shown in both the daily and triweekly probiotic groups (adjusted IRR 0.83, 95% CI 0.72-0.96 in both groups) and during T0-T12 (adjusted IRR 0.84, 95% CI 0.76-0.94, and adjusted IRR 0.86, 95% CI 0.76-0.96, respectively) compared to the placebo group. In contrast, significantly increased regressive surfaces were demonstrated during T0-T6 by adjusted IRR 1.76 (95% CI 1.25-2.48) and 2.01 (95% CI 1.42-2.85) and during T0-T12 by adjusted IRR 1.98 (95% CI 1.50-2.61) and 1.92 (95% CI 1.45-2.55) for the daily and triweekly groups, respectively. In conclusion, probiotic milk consumption either daily or triweekly can modestly prevent new caries, but considerably reverse carious lesions, suggesting that a daily or triweekly dose interval is sufficient to reverse carious lesions.

Reduction of Streptococcus mutans by probiotic milk: a multicenter randomized controlled trial.

OBJECTIVE: To investigate the effects of probiotics, Lactobacillus paracasei SD1, on the quantities of Streptococcus mutans in saliva and plaque samples of preschool children. DESIGN: This randomized trial recruited 487 preschool children from eight childcare centers. Participants were assigned to receive a 6-month course of placebo milk daily (group I), probiotic milk either daily (group II) or three days a week (triweekly, group III). The absolute quantities of S. mutans and total lactobacilli in the saliva and plaque samples at baseline (T0), after intervention (T6), and 6 months after discontinuation (T12) were assessed by qPCR. RESULTS: Of 487 children, 354 completed all follow-up periods. However, only 268 children (3.2 ± 0.8 years old; groups I = 86, II = 89, and III = 93) provided adequate saliva for qPCR. Whereas the quantities of S. mutans were significantly decreased in groups II and III compared to group I in the saliva and plaque samples at T6 and T12, those of total lactobacilli were significantly increased (p < 0.0167). There was no difference in the quantities of S. mutans or total lactobacilli between groups II and III at any period. Significant changes in the quantities of S. mutans and total lactobacilli lasted until T12 compared to T0 (p < 0.0167). CONCLUSIONS: Probiotic administration daily or triweekly reduces S. mutans quantities, whereas it increases total lactobacilli quantities that persists at least 6 months after discontinuation in the saliva and plaque samples of preschool children. CLINICAL RELEVANCE: Daily or triweekly consumption of L. paracasei SD1 supplemented in milk may help prevent dental caries in preschool children.

Preaxial polydactyly associated with a MSX1 mutation and report of two novel mutations.

We report two novel heterozygous missense MSX1 mutations in two Thai families (c.739C>T; p.Pro247Ser and c.607G>A; p.Ala203Thr). The p.Ala203Thr mutation was found in a female patient, her sister, and their father and is associated with unilateral cleft lip and palate, hypodontia, and microdontia. The p.Pro247Ser mutation was found in a three-generation Thai family and was associated with bilateral cleft lip and palate, hypodontia, microdontia, and dens invaginatus. The proband also had preaxial polydactyly of the left hand. The role of Msx1 in limb development in mice is discussed. Intrafamilial variability of the phenotypes is clearly evident. This is the first time that a limb anomaly has been reported to be associated with a mutation in MSX1.

Enhancement of salivary human neutrophil peptide 1-3 levels by probiotic supplementation.

BACKGROUND: Probiotic supplementation can reduce mutans streptococci (MS) numbers. One of its proposed mechanisms is immunomodulation. Salivary human neutrophil peptide 1-3 (HNP1-3) levels have previously been demonstrated to be higher in caries-free than in caries-susceptible children, suggesting their preventive role against caries. We aimed to compare salivary HNP1-3 levels between an intervention group with probiotics and a control group. METHODS: A randomized double-blinded clinical trial was conducted. Sixty schoolchildren were equally allocated to either an intervention or control group. The use of a probiotic strain, Lactobacillus paracasei SD1, has shown to reduce MS numbers in volunteers. In unstimulated whole saliva, HNP1-3 levels were assayed by ELISA, and MS and lactobacilli counts were assayed by colony counting at baseline (T0) and at 3 (T3), 6 (T6), and 12 months (T12). The International Caries Detection and Assessment system was used to assess caries status. RESULTS: In the intervention group, salivary HNP1-3 levels were significantly greater than those in the control group at T3 and T6 (p < 0.001), whereas MS counts were significantly decreased (p < 0.01). In the intervention group, positive and negative correlations were found between HNP1-3 levels and lactobacilli counts and between MS and lactobacilli counts, respectively. However, there was no significant correlation between enhanced HNP1-3 levels and decreased MS numbers. The caries increment for the pit and fissure surface, but not for the smooth surfaces, was significantly decreased in the intervention group compared with the control group (p = 0.01). CONCLUSIONS: Probiotics can temporarily enhance salivary HNP1-3 levels; however, their action to reduce new pit and fissure caries probably involves microbial interactions. TRIAL REGISTRATION: TCTR20130904001 (registration date: September 04, 2013).