Severe Early Childhood Caries Group Research Articles

Quantitative Analysis of Bifidobacterium and Scardovia wiggsiae in Dental Plaque from Children in Northern Thailand and Their Association with Caries Factors

Abstract Objectives The aim of this study were to measurably detect Bifidobacterium and Scardovia wiggsiae in plaque from severe early childhood caries (S-ECC) and caries-free children and to analyze the interrelation between these bacteria and clinical caries presentation and caries-related factors assessed by questionnaire. Materials and Methods One-hundred forty supra gingiva plaque samples from children aged between 2 and 5 years were used in this study. There were 70 children in each group. Recorded plaque index, modified gingival index, and decay, missing, and filled tooth (dmft) scores. Parents' attitudes, child's oral hygiene, and diet were assessed by questionnaire. DNA was extracted from plaque samples and quantitative real-time polymerase chain reaction using fluorescent dye was performed. Results Plaque (p < 0.001) and modified gingival indices (p < 0.001) in the S-ECC group were higher than in the caries-free group. Prevalence of Bifidobacterium (p = 0.004) and S. wiggsiae (p < 0.001) in the S-ECC group was higher than in the caries-free group. The numbers of total bacteria (p = 0.003), Bifidobacterium (p < 0.001), and proportion of Bifidobacterium to total bacteria (p < 0.001) were higher in the S-ECC group. Detections of both bacteria, Bifidobacterium + S. wiggsiae (p < 0.001), were higher in the S-ECC group than in the caries-free group. In the S-ECC group, dmft scores (p < 0.001; p = 0.024) and the modified gingiva index (p = 0.004; p = 0.002) were higher in the presence of Bifidobacterium and S. wiggsiae, respectively. In the S-ECC group, the dmft scores (p = 0.005) and modified gingiva index (p = 0.004) were higher in the presence of both Bifidobacterium + S. wiggsiae. There were positive correlations between the Bifidobacterium level (p = 0.003), the proportion of Bifidobacterium to total bacteria (p = 0.017), and S-ECC. The level of Bifidobacterium (p < 0.001) and ratio of Bifidobacterium to total bacteria (p < 0.001) were correlated with the dmft score and modified gingival index in the S-ECC group. From the questionnaire, S-ECC were associated with major caregiver (p = 0.002), parent education levels (p = 0.02), prolonged bottle-feeding (>18 months) (p = 0.015), night-time feeding (p < 0.001), eating cariogenic snacks (p = 0.019), and frequency of dental visits (p = 0.003). Conclusions Levels of total bacteria, Bifidobacterium, S. wiggsiae and plaque, and the modified gingival indices were higher in the S-ECC group. Factors associated with S-ECC included the major caregiver, parent education levels, feeding patterns, cariogenic snacks consumption, and frequency of dental visits.

Relationship between parental adverse childhood experiences and the prevalence of early childhood caries.

The aim of this study was to examine the behavioural health conditions associated with parents' retrospective adverse childhood experiences (ACEs) scores and their children's early childhood caries (ECC) in parent-child dyads. Parents with children younger than 72 months were included in the study. A relational screening model was used. Interaction among ACEs, ECC, nutritional habits and oral hygiene habits were evaluated. Chi-square tests and t-tests were used in the study. Multiple variables were evaluated using the artificial neural network (ANN) model. The mean age of the 535 children included in the study was 46.5 months, and 52% were female. Using the ANN model, there was a statistically significant relationship between the educational status of the mothers in both the ECC and severe ECC (S-ECC) groups and the socioeconomic status of the family (p < 0.05). If the number of snacks consumed daily was three or more, the risk of ECC was statistically significantly higher (chi-square test p = 0.034). The parents' ACEs scores had an impact on both ECC and S-ECC formation (p = 0.001, t-test). The higher the ACEs score, the higher the risk of S-ECC. The mean ACEs scores of the parents were also significantly higher in both the ECC and S-ECC groups compared to those of the parents of children without dental caries (p = 0.001, t-test). It was calculated that ACEs scores were effective at a rate of 18.2% on ECC (p = 0.045, ANN). The ACEs scores of parents have an impact on the oral health of young children and ECC/S-ECC formation. The long-term effects of parental ACEs are reflected in their children's oral health. Therefore, reducing the psychosocial determinants ACEs and providing parental support may help in overcoming barriers to the well-being of young children and may facilitate better oral health.

Relationship between the salivary concentrations of proteinase-3 and interleukin-8 and severe early childhood caries.

Severe early childhood caries (S-ECC) is a multifactorial transmissible infectious disease continuing to affect infants and toddlers worldwide. Saliva plays a modulatory role in the pathogenesis of dental caries. The present study aimed to assess the salivary levels of proteinase-3 (PR3) and interleukin-8 (IL-8) as pro-inflammatory cytokines related to the function of neutrophils in association with S-ECC and its treatment. Fifty children aged 36-60 months were recruited (25 caries-free controls and 25 S-ECC patients). Saliva sampling was performed in all participants. In the S-ECC group, sampling was repeated 6-8 weeks after restorative treatment. The salivary concentrations of PR3 and IL-8 were determined using the enzyme-linked immunosorbent assay (ELISA). The χ2 test, Fisher's exact test, the independent t test, and the paired t test were applied at p < 0.05. The baseline salivary concentrations of PR3 and IL-8 in the S-ECC group were significantly higher than in the caries-free group (p < 0.001). A significant reduction occurred in the levels of these cytokines following restorative treatment in the S-ECC group (p < 0.001), although they were still significantly higher than in the caries-free group (p < 0.05). The salivary levels of PR3 and IL-8 were significantly affected by the presence of dental caries in children, implying their potential efficiency as non-invasive indicators in the determination of the caries risk and treatment effectiveness.

Relationship between the salivary concentration of matrix metalloproteinases 8 and 20 and severe early childhood caries.

Dental caries is initiated through mineral dissolution by bacterial acids and collagen degradation by endogenous proteolytic enzymes, mainly collagenolytic matrix metalloproteinases (MMPs). The present research aimed to evaluate the relationship between severe early childhood caries (S-ECC) and salivary MMP-8 and MMP-20 concentrations. Fifty children aged 36-60 months were assigned to either the caries-free (control) group or the S-ECC group. Standard clinical examinations were performed, and approx. 1 mL of expectorated unstimulated whole saliva was collected from all participants. In the S-ECC group, the sampling was repeated 3 months after restorative treatment. All samples were analyzed for the salivary concentrations of MMP-8 and MMP-20, using the enzyme-linked immunosorbent assay (ELISA). Statistical analysis employed the t test, the Mann-Whitney U test, the χ2 test, Fisher's exact test, and the paired samples t test. The level of significance was set at 0.05. At baseline, the subjects in the S-ECC group presented with significantly elevated levels of MMP-8 as compared to the control group. However, the salivary concentration of MMP-20 did not exhibit a significant difference between the 2 groups. A significant reduction occurred in the levels of MMP-8 and MMP-20 3 months after restorative treatment in the S-ECC group. The salivary levels of MMP-8 and MMP-20 were significantly affected by dental restorative treatment in children. Furthermore, MMP-8 was observed to be a better indicator of the dental caries status than MMP-20.

Salivary Leptin Levels in Children with Early Childhood Caries – An Interventional Study

Background: Early childhood caries (ECC) is a pathologic condition of multifactorial nature. The diagnosis of the same has been limited to clinical and radiographic aids with very little significance to assess its inflammatory component. objectives: The present study aims to evaluate the role of leptin, an adipocytokine as a non-invasive inflammatory biomarker in ECC. Materials and Method: A total of 60 children between the age of (3-5) years were selected for the study. The study subjects were divided into three groups of 20 each including Group 1 as control, Group 2 with mild to moderate ECC and Group 3 with severe ECC. Saliva samples were taken from all subjects and collected again after 2 months following rehabilitative intervention. Levels of salivary leptin were determined using Enzyme-Linked Immunosorbent Assay (ELISA). Results: Levels of salivary leptin were significantly associated with severity of dental caries. The intragroup comparisons of pre and post treatment levels of salivary leptin showed significant reductions in both mild to moderate ECC and severe ECC groups following caries control. Inter group evaluation between mild to moderate ECC and severe ECC post treatment showed statistically significant decline of leptin levels in comparison to baseline values. Conclusion: There was a statistically significant decline in salivary leptin levels between the mild moderate and severe ECC group, after 2 months following caries control. Thus, leptin holds a potential to be recognized as a reliable future prognostic and diagnostic inflammatory marker in early childhood caries.

Identification of Scardovia Wiggsiae in Children Between Severe Early Childhood Caries and Caries-Free Children – A Polymerised Chain Reaction -Based Study

Dental caries is the most common microbial disease of oral cavity, affecting 60 – 90% of children worldwide. Early Childhood Caries (ECC) is a major health problem in developing countries. These lesions are destructive and can affect the development of permanent successors. Although Streptococcus mutans is the primary cause of ECC, some studies support the view that caries can develop in the absence of mutans streptococci. Scardovia wiggsiae, a gram-positive anaerobe was recently found to be associated with Early Childhood Caries in the presence and absence of Streptococcus mutans. The aims were to investigate the cariogenic potential of S. wiggsiae, in ECC. The study was intended to compare the presence of S. wiggsiae, in the saliva samples of caries-free children and children with severe ECC, thereby determine this new acidogenic species noxiousness as a pathogen in Early Childhood Caries.Saliva samples were collected from 40 children, age ranging between three to six years. The sample thus collected at room temperature was then subjected to DNA extraction and Polymerase Chain Reaction (PCR) analysis to evaluate the presence of S. wiggsiae. Numerical parameters were tested for significance using Fisher’s Exact Test and the statistical analysis using Epi Info software.The tested samples revealed almost equal proportion of Scardovia wiggsiae in caries-free group and severe ECC group. S. wiggsiae was isolated and detected in both the study and the control groups. Statistical significance was not pronounced claiming the falsehood of the association between Early childhood caries and Scardovia wiggsiae.

Determination of salivary concentrations of leptin and adiponectin, ability to reduce ferric ions and total antioxidant capacity of saliva in patients with severe early childhood caries.

IntroductionOne of the most common oral diseases affecting children is early childhood caries (ECC). The link between oxidative stress and ECC has been proven in numerous clinical studies. Technical and biological variability were so high in most of the studies that none of the markers have yet been proven suitable for routine clinical use. This study aimed to evaluate the antioxidant status and the levels of leptin and adiponectin in saliva of children with severe early childhood caries (S-ECC).MethodsMorning unstimulated saliva samples were collected from children (n = 40, 0–6 years old) for the evaluation of oxidative stress which were measured by total antioxidant capacity (TAC), and by the ferric reducing antioxidant power (FRAP) assays, as well as to assess the salivary levels of leptin and adiponectin. FRAP, TAC, leptin and adiponectin concentrations were evaluated in S-ECC group (n = 31) and caries free group CF (n = 9). All results were analyzed based on age and sex.ResultsOverall median salivary leptin and adiponectin levels were 5.59 pg/mL and 24.86 ng/mL, respectively. Significantly lower leptin levels were observed in saliva of caries free children (4.66 pg/mL) than in the S-ECC group (6.64 pg/mL, p < 0.01). No significant difference was observed for adiponectin levels (S-ECC and CF, 25.31 and 23.2 ng/mL, respectively, p = 0.961). TAC and FRAP values of saliva had similar values in children with S-ECC and caries free children. TAC and FRAP values also remained stable with the age of the children, without significant differences with respect to sex.ConclusionThe increased concentrations of leptin in saliva of children with S-ECC suggests that leptin may play a role in inflammatory and immune responses in the development of early childhood caries.

Comparative real-time PCR quantification of cytomegalovirus in severe early childhood caries and caries-free children

The present study was undertaken to evaluate cytomegalovirus (CMV) load in carious lesions in children with severe early childhood caries (S-ECC) and compare it with caries-free (CF), healthy children, by quantitative real-time polymerase chain reaction (PCR). Dental plaque samples were collected from 57 subjects including S-ECC (n = 33) and CF (n = 24) patients. The two groups were matched in terms of age, gender, and socioeconomic status. After deoxyribonucleic acid (DNA) extraction, SYBR Green real-time PCR was used to quantify CMV DNA. The CMV DNA was detected in seven patients in theS-ECC group. The mean of decayed, missing, and filled tooth surfaces (dmfs) was 11.90 ± 5.95 in the S-ECC group. There was no significant difference in gender between the two groups. There was a significant relationship between CMV positivity in children and theS-ECC group (p = 0.043). Results of this study suggest a relationship between the presence of CMV in the oral cavity and S-ECC. We can design clinical trials to confirm these results and improve children's dental care.

Streptococcus mutans-associated bacteria in dental plaque of severe early childhood caries

ABSTRACT Background Streptococcus mutans (S. mutans) is a potential pathogenic bacteria of dental caries. However, the level of S. mutans is low in some children with severe early childhood caries (SECC) Aim To evaluate the effect of S. mutans level on dental microbiome and cariogenesis. Methods The oral microbiota was compared between caries-free group (CF) and SECC group.16S rRNA gene sequencing was used for S. mutans level bacterial community analysis. The candidate bacteria that were closely related with S. mutans abundance were identified and confirmed by absolute quantitative real-time PCR in clinical dental plaque samples from CF and SECC groups. Results Through in-depth analysis of dental plaque microorganism, Leptotrichia, Selenomonas and Prevotella_7 were found in the S. mutans-low group (p < 0.05) and Porphyromonas, Selenomonas_3 were found in the S. mutans-high group (p < 0.05). Through quantitative real-time PCR, Leptotrichia, Selenomonas and Prevotella_7 were identified as the potential biomarkers of SECC when S. mutans was at a low level. Conclusion Leptotrichia, Selenomonas and Prevotella_7 are identified as potential biomarkers in SECC with a low abundance or without S. mutans. Our study may shed light on the understanding of caries occurrence in SECC with low abundance of S. mutans. Abbreviations S. mutans, Streptococcus mutans; CF, caries-free; SECC, severe early childhood caries; ECC, early childhood caries; rRNA, ribosome RNA; qPCR, Quantitative real-time PCR; OTUs, operational taxonomic units; ANOVA, analysis of variance; LDA, Linear discriminant analysis; LEfSe, Linear discriminant analysis effect size; COG, Groups of proteins; NMDS, Non-MetricMulti-Dimensional Scaling; IL-1β, interleukin −1β; IL-6, interleukin-6; IL-8, interleukin-8; IL-10, interleukin-10.

Evaluation of salivary thiobarbituric acid reactive substances, total protein, and pH in children with various degrees of early childhood caries: a case-control study.

According to new studies, oxidative stress may play an important role in the beginning and progression of oral diseases. The aim of this study was the evaluation of thiobarbituric acid reactive substances (TBARS) as an indicator of lipid peroxidation, total protein, and pH of saliva in children with various degrees of early childhood caries in comparison with caries-free children. This case-control study was carried out on 90 children 3-5years old in three equal groups, including caries-free children (CF), early childhood caries (ECC), and severe early childhood caries (S-ECC) who were age- and sex matched. Saliva pH was measured using a paper pH meter and TBARS and total protein were measured by spectrophotometer methods and data were statistically analysed. TBARS and total protein levels in ECC and S-ECC groups were significantly higher than the CF group (p < 0.001), but pH was not significantly different between groups. There were no significant differences between ECC and S-ECC groups in any of the studied indicators. Considering the limitations of the present study, the results showing the higher salivary TBARS levels in the S-ECC and ECC groups compared to caries-free children, lead to the conclusion that oxidative stress could be considered as a risk factor for early childhood caries.

Correlation of Streptococcus Mutans by 16SrRNA Gene Sequencing in Children with Different Caries Experience- A PCR Based Study

Background and Objectives: Early Childhood Caries is considered as a common chronic disease in children. Since some research had concluded that children with higher levels of Streptococcus mutans are associated with a higher incidence of decayed, missing, and filled teeth, the present study was undertaken to analyze the salivary Streptococcus mutans of children with different caries status using species specific 16S rRNA gene sequencing technology to evaluate the quantity of Streptococcus mutans with respect to different caries status.&#x0D; Materials and Methods: Children between 3-6 years were selected and divided into 3 groups, Group I- Caries free, Group II-Early Childhood caries and Group III- Severe Early childhood caries. The caries status was assessed using dmfs and the severity of caries was assessed using pufa index. Salivary samples were collected to isolate DNA and Real time PCR was done with 16SrRNA primer specific to Streptococcus mutans to estimate the quantity of Streptococcus mutans in children with different caries status.&#x0D; Results: There was a significant difference in the mean CT values among the study groups (p &lt; 0.001). Post hoc Tukey test revealed that, Group I had a significantly higher mean CT values than that of Groups II and III. Pearson's correlation analysis was carried out to correlate dmfs value with CT values and to correlate the pufa value with the fold change. There was a negative correlation between dmfs score and CT values and positive correlation between pufa score and fold change.&#x0D; Conclusion: There was a higher expression of 16SrRNA gene in Severe Early childhood caries group, followed by Early childhood caries and caries free group, indicating a high level of Streptococcus mutans in Severe Early childhood caries group and there was a negative correlation between dmfs score and CT values and a positive correlation between pufa score and fold change.

Metaproteomics associated with severe early childhood caries highlights the differences in salivary proteins

ObjectiveTo investigate the salivary metaproteomic characteristics of the children with and without severe early childhood caries (S-ECC). DesignIn this study, we collected unstimulated saliva samples from 34 children (age 3–4 years) with caries free (NC, dmfs (= index of decayed, missing due to caries, or filled tooth surfaces) = 0, n = 23) and with S-ECC (dmfs≥10, n = 11). Salivary proteins were extracted and reduced, and then a Liquid Chromatography/Mass Spectrometry system was used to identify proteins. ResultsNearly 3000 proteins were identified in this study, and about 3.5 % of the proteins originated from human while 86 % were derived from microbes. The salivary protein types in the NC group were statistically greater than those in the S-ECC group (P <0.05). Specifically, the salivary protein types derived from microbes in the NC group were significantly greater than those in the S-ECC group. Three proteins, human lactoferrin, penicillin-binding protein 1C [Burkholderia ubonensis], human alpha-defensin 1 (F28a mutant), were decreased statistically in the NC group compared to the S-ECC group (P < 0.05). Only one protein, 50S ribosomal protein L17 secreted by Haemophilus haemolyticus, was significantly increased in the NC group compared to the S-ECC group. Salivary IgA was the top highest protein in the NC group whereas human lysozyme was the top highest protein in the S-ECC group. ConclusionsThe differential proteins recognized in this study may be conducive for finding a caries biomarker. Understanding the metaproteomic characteristics can help us to control the caries from human origin and microbial origin.

Characterization of Supragingival Plaque and Oral Swab Microbiomes in Children With Severe Early Childhood Caries.

The human oral cavity harbors one of the most diverse microbial communities with different oral microenvironments allowing the colonization of unique microbial species. This study aimed to determine which of two commonly used sampling sites (dental plaque vs. oral swab) would provide a better prediction model for caries-free vs. severe early childhood caries (S-ECC) using next generation sequencing and machine learning (ML). In this cross-sectional study, a total of 80 children (40 S-ECC and 40 caries-free) < 72 months of age were recruited. Supragingival plaque and oral swab samples were used for the amplicon sequencing of the V4-16S rRNA and ITS1 rRNA genes. The results showed significant differences in alpha and beta diversity between dental plaque and oral swab bacterial and fungal microbiomes. Differential abundance analyses showed that, among others, the cariogenic species Streptococcus mutans was enriched in the dental plaque, compared to oral swabs, of children with S-ECC. The fungal species Candida dubliniensis and C. tropicalis were more abundant in the oral swab samples of children with S-ECC compared to caries-free controls. They were also among the top 20 most important features for the classification of S-ECC vs. caries-free in oral swabs and for the classification of dental plaque vs. oral swab in the S-ECC group. ML approaches revealed the possibility of classifying samples according to both caries status and sampling sites. The tested site of sample collection did not change the predictability of the disease. However, the species considered to be important for the classification of disease in each sampling site were slightly different. Being able to determine the origin of the samples could be very useful during the design of oral microbiome studies. This study provides important insights into the differences between the dental plaque and oral swab bacteriome and mycobiome of children with S-ECC and those caries-free.

Comparison of Salivary Protein Profile and Dietary Habits in Children With and Without Early Childhood Caries

Background and objectives: The purpose of this study was to evaluate the relationship between the total salivary protein concentration and dietary habits in Caries-free, Early childhood Caries and Severe Early Childhood Caries children.&#x0D; Materials and methods: The present study included Thirty children who were divided into three groups. They were further divided into Caries-free, Early childhood Caries and Severe Early Childhood Caries with 10 children in each group. Information was collected regarding oral hygiene practices, feeding habits and dietary pattern through a face-to-face interview with the parent/caretaker of the child. Unstimulated saliva was collected and total protein concentration was estimated. Multivariate logistic regression analysis was performed to assess the relative contributions of dietary and feeding patterns between children with and without ECC. ANOVA was done to identify the difference in mean protein concentration between the groups.&#x0D; Results: Risk factors specific to the ECC group were Breast feeding &lt;24 months (OR 2.1, 95% CI, 0.35-13.0), bottle feeding (21.0, 95% CI, 2.8-15.7), snacking more than 3 times per day (OR 1.22, 95% CI 0.283-5.734), consumption of less protein diet (OR 4.1, 95% CI, 1.99-8.81), consumption of less milk products (2.33, 95% CI, 1.67-6.77). Results also revealed that there was no statistically significant difference in total protein concentration between Caries-free, Early childhood Caries and Severe Early Childhood Caries group.&#x0D; Conclusion: Within the limitation of this study, we can conclude that, Breast feeding less than 24 months, bottle feeding, snacking more than thrice per day, consuming low protein diet and low milk products had a high risk of developing ECC. There was no significant difference in the mean protein concentration between caries free, ECC and S ECC group.

Translation and Validation of Oral Health-Related Early Childhood Quality of Life Tool for Nepalese Preschool Children

Background Early childhood caries (ECC) in preschool children debilitate their quality of life affecting oral function, appearance and social well-being. A tool for measuring oral health-related quality of life for preschoolers may help in making clinical decisions and assist policy makers in planning and management of health programs in the country.&#x0D; Objective To translate oral health-related early childhood quality of life (OH-ECQoL) tool into Nepali language and test its validity and reliability.&#x0D; Method OH-ECQoL was translated into Nepali language by two Nepalese professional translators which was back translated by professional English translator. This was sent to three independent advisors to see the appropriateness of translation. Pilot testing was done in 20 parents and questionnaire was finalized after needed corrections. Final version was introduced to the 118 parents of children aged 24- 71 months. Caries status was recorded for all children participating in the study. Concurrent, construct, discriminant validity and internal consistency reliability, test -retest reliability were evaluated.&#x0D; Result OH-ECQoL scores and perception of parents for general and oral health of their children was significant (at 0.01 level). There was significant difference in OH-ECQoL scores between no ECC and severe ECC groups and moderate and severe ECC groups (at 0.05 level). There was also a significant correlation between child impact section and family impact section (at 0.01 level). Cronbach’s alpha was 0.891 demonstrating good internal consistency. Intra class coefficient was 0.963 suggesting excellent test-retest reliability. Ninety-one (77.1%) children had severe ECC and 40(33.9%) parents were from upper middle class.&#x0D; Conclusion The Nepali version of OH-ECQoL is a valid and reliable tool for assessing the oral health-related early childhood quality of life in children of Nepal.

The association between the salivary cortisol level of mothers, children's temperament, and early childhood caries.

Multiple factors can affect early childhood caries (ECC). Maternal stress and child's temperament, as predictors of child behavior, are among factors that may be associated with ECC. The aim of this study was to evaluate the association between the salivary cortisol level of mothers as an indicator of maternal stress, child's temperament, and ECC. Ninety preschool children, who were divided into three groups of ECC, severe ECC (S-ECC), and caries-free (CF), were included in this study, as well as their mothers with no history of anxiety disorders. The salivary cortisol levels of mothers were analyzed using ELISA assay. Children were examined for the evidence of caries, using the decayed-missing-filled teeth index, based on the World Health Organization standard criteria. Child temperament was also assessed, using Cloninger's Preschool Temperament and Character Inventory, consisting of seven significant factors (cooperativeness, harm avoidance, novelty seeking, reward dependence, persistence, self-directing, and self-transcendence). The association between the salivary cortisol level of mothers and temperament of children with S-ECC was positive although it was not statistically significant (P > 0.05). Of seven temperament factors, only harm avoidance was significantly different between the CF and S-ECC groups (P = 0.016). Maternal stress and child's temperament were not associated with ECC. Based on the present findings, children with less harm avoidance may be more prone to S-ECC.

The enhancing antibiofilm activity of curcumin on Streptococcus mutans strains from severe early childhood caries

BackgroundStreptococcus mutans (S. mutans) is one of the main cariogenic bacteria for caries. It was found that the clinical strains of S. mutans isolated from caries active population have stronger cariogenic ability than the isolates from caries-free (CF) people. Previous studies have found that curcumin can inhibit biofilm formation of S. mutans UA159. The objective of this study is to explore the antibiofilm effect of curcumin on the clinical isolates of S. mutans from severe early childhood caries(SECC).ResultsThe isolates from SECC group had more biomass than CF group (t = 4.296, P < 0.001). The acidogenicity and aciduricity of the strains from two groups showed no significant difference. After treatment with curcumin, the viability of biofilm was reduced to 61.865% ± 7.108% in SECC and to 84.059% ± 10.227% in CF group at 24 h (P < 0.05). The net reduction of live bacteria and total bacteria in the SECC group was significantly higher than that of the CF group (live bacteria t = 3.305, P = 0.016; total bacteria t = 2.378, P = 0.045) at 5 min. For 24 h, the net reduction of live bacteria and total bacteria in the SECC group was significantly higher than that of the CF group (live bacteria t = 3.305, P = 0.016; total bacteria t = 2.378, P = 0.045). The reduction of biofilm thickness reduced significantly in 5 min (t = 4.110, P = 0.015) and in 24 h (t = 3.453, P = 0.014). Long-term (24 h) curcumin treatment inhibited the amount of EPS in SECC group from (25.980 ± 1.156) μm3/μm2 to (20.136 ± 1.042) μm3/μm2, the difference was statistically significant (t = 7.510, P < 0.001). The gene of gtfC, gtfD, ftf, gbpB, fruA and srtA in the CF group and the gtfB, gtfC, gtfD, ftf, gbpB, srtA in SECC group were respectively reduced after 5 min curcumin treatment. After 24 h treatment, the gtfB, gtfC, gtfD, ftf, gbpB, fruA and srtA in both two groups were downregulation, all the differences were statistically significant.ConclusionsCurcumin has antibiofilm activity on clinical strains of S. mutans, especially for those isolated from SECC.

Quantitative Analysis of Salivary Oral Bacteria Associated with Severe Early Childhood Caries and Construction of Caries Assessment Model

To construct a saliva-based caries risk assessment model, saliva samples from 176 severe early childhood caries (S-ECC) children and 178 healthy (H) children were screened by real-time PCR-based quantification of the selected species, including Streptococcus mutans, Prevotella pallens, Prevotella denticola and Lactobacillus fermentum. Host factors including caries status, dmft indices, age, gender, and geographic origin were assessed in their influence on abundance of the targeted species, which revealed host caries status as the dominant factor, followed by dmft indices (both P < 0.01). Moreover, levels of S. mutans and P. denticola in the S-ECC group were significantly higher than those in the healthy group (P < 0.001 for S. mutans and P < 0.01 for P. denticola). Interestingly, the co-occurrence network of these targeted species in the S-ECC group differed from that from the healthy group. Finally, based on the combined change pattern of S. mutans and P. pallens, we constructed an S-ECC diagnosis model with an accuracy of 72%. This saliva-based caries diagnosis model is of potential value for circumstances where sampling dental plague is difficult.

Evaluation of oral streptococci in saliva of children with severe Early Childhood Caries and caries-free.

Oral streptococci were found to be associated with Early Childhood Caries. The aim of this study was to evaluate the 6 different bacteria in the streptococcus group in the saliva of children with severe early childhood caries (S-ECC) by polymerase chain reaction (PCR). A total of 60 children between 3 and 6 years of age were divided into two groups: children with S-ECC (Group S-ECC; n=30) and children who were caries-free (Group CF; n=30), according to the dmft and dmfs indices. Unstimulated saliva was collected from all participants for the detection of streptococcal group bacteria, including: Streptococcus mutans, Streptococcus oralis, Streptococcus sanguinis, Streptococcus gordonii, Streptococcus salivarius, and Streptococcus sobrinus, using PCR-restriction fragment length polymorphism (RFLP) of amplified 16S rRNA gene. The data were analysed using SPSS software. The prevalence of S. oralis was significantly higher in the S-ECC group compared to the CF group (p<0.05). However, the frequencies of S. mutans, S. sanguinis, S. gordonii, S. salivarius, and S. sobrinus were similar between the two groups (p>0.05). The amount of streptococci colonies was higher in the S-ECC group compared to the CF group (p<0.05). S. mutans or S. sobrinus alone may not be the only indicators for high risk of caries, but the prevalence of S. oralis in saliva may be a risk factor for increased caries activity in S-ECC.

Sex-Based Diverse Plaque Microbiota in Children with Severe Caries

Severe early childhood caries (S-ECC) is a multifactorial disease that can lead to suffering and reduced oral health–related quality of life in young children. The bacterial and fungal composition of dental plaque and how children’s sex is associated with S-ECC are largely unknown. In this study, V4-16S rRNA and ITS1 rRNA gene amplicon sequencing was used to compare the plaque bacteriome and mycobiome of children <72 mo of age: 40 with S-ECC (15 males, 25 females) and 40 caries-free (19 males, 21 females). Health- and nutrition-related questionnaire data were also investigated. This study aimed to analyze potential sex-based differences in the supragingival plaque microbiota of young children with S-ECC and those caries-free. Behavioral and nutritional habit differences were observed between children with S-ECC and those caries-free and between male and female children. Overall, higher levels of Veillonella dispar, Streptococcus mutans, and other bacterial species were found in the S-ECC group as compared with caries-free controls (P < 0.05). A significant difference in the abundance of Neisseria was observed between males and females with S-ECC (P < .05). Fungal taxonomic analysis showed significantly higher levels of Candida dubliniensis in the plaque of children with S-ECC as compared with those caries-free (P < 0.05), but no differences were observed with Candida albicans (P > 0.05). Significant differences in the relative abundance of Mycosphaerella, Cyberlindnera, and Trichosporon fungal species were also observed between the caries-free and S-ECC groups (P < 0.05). Machine learning analysis revealed the most important bacterial and fungal species for classifying S-ECC versus caries-free. Different patterns of crosstalk between microbial species were observed between male and female children. Our work demonstrates that plaque microbiota and sex may be important determinants for S-ECC and could be factors to consider for inclusion in caries risk assessment tools.