Metabolomic analysis identifies differentially produced oral metabolites, including the oncometabolite 2-hydroxyglutarate, in patients with head and neck squamous cell carcinoma.

BACKGROUND: Metabolomics represents a promising approach for discovering novel targets and biomarkers in head and neck squamous cell carcinoma (HNSCC). Here we used metabolomics to identify oral metabolites associated with HNSCC. METHODS: Tumor and adjacent normal tissue from surgical resections and presurgical oral washes as well as oral washes were collected from healthy participants. Metabolites extractions of these samples were analyzed by liquid chromatography-mass spectroscopy (LC/MS), LC/MS/MS and gas chromatography-MS (GC/MS). RESULTS: Among 28 samples obtained from 7 HNSCC cases and 7 controls, 422 metabolites were detected (269 identified and 153 unidentified). Oral washes contained 12 and 23 metabolites in healthy controls and HNSCC patients, respectively, with phosphate and lactate being the most abundant. Small molecules related to energy metabolism were significantly elevated in HNSCC patients compared to controls. Levels of beta-alanine, alpha-hydroxyisovalerate, tryptophan, and hexanoylcarnitine were elevated in HNSCC oral washes compared to healthy controls (range 7.8-12.2-fold). Resection tissues contained 22 metabolites, of which eight were overproduced in tumor by 1.9- to 12-fold compared to controls. TCA cycle analogs 2-hydroxyglutarate (2-HG) and 3-GMP were detected exclusively in tumor tissues. Targeted quantification of 2-HG in a representative HNSCC patient showed increase in tumor tissue (14.7 µg/mL), but undetectable in normal tissue. Moreover, high levels of 2-HG were detected in HNSCC cell lines but not in healthy primary oral keratinocyte cultures. CONCLUSIONS: Oral metabolites related to energy metabolism were elevated in HNSCC, and acylcarnitine and 2HG may have potential as non-invasive biomarkers. Further validation in clinical studies is warranted.

Association of blood lead levels in children 0-72 months with living in Mid-Appalachia: a semi-ecologic study.

INTRODUCTION: Lead exposure in children remains a significant public health issue, although many advances have been made. The Mid-Appalachia area (Kentucky, New York, Ohio, Pennsylvania, and West Virginia) is 89-91% rural with a population density of 16-21 people/km2 (41-54 people/mi2). Mid-Appalachia has significant health disparities including concerns for the consequences of greater lead exposure to children due to mining and industrial footprints, and existing older housing. The purpose of this study is to compare the reported blood lead levels of screened children, aged 0-72 months in Mid-Appalachia, to the children in the USA in general. METHODS: Data from the Centers for Disease Control and Prevention and from the US Census Bureau were analyzed in a semi-ecological study. The blood lead level of 5 µg/dL was compared between children in Mid-Appalachia and the US housing units built before 1950; US housing units built before 1940 were also compared. RESULTS: The number of children with blood lead levels of 5 µg/dL was significantly greater in Mid-Appalachia than nationally (7.75% vs 5.79%, respectively; p<0.0001). The number of homes built before 1950 (p<0.0001) and built before 1940 (p<0.0001) was significantly greater in Mid-Appalachia than nationally. CONCLUSIONS: Blood lead levels in children are higher in Mid-Appalachia than nationally and there is an ecological relationship with the number of homes built before 1950 and before 1940.

Association of Alanine Aminotransferase and Periodontitis: A Cross-Sectional Analysis-NHANES 2009-2012.

Objective. Alanine Aminotransferase is an enzyme associated with not only liver diseases, liver conditions, and metabolic syndrome, but also inflammation. Periodontitis is associated with increased cytokines and other markers of inflammation. The purpose of this study is to determine if an independent association between Alanine Aminotransferase and periodontitis exists. Methods. Data from the 2009-2010 and 2011-2012 National Health and Nutrition Surveys (NHANES) were combined. Data concerning periodontitis and Alanine Aminotransferase were extracted and analyzed with Rao Scott Chi-square and logistic regressions. Serum Alanine Aminotransferase was dichotomized at 40 units/liter, and periodontitis was dichotomized to the presence or absence of periodontitis. Results. In bivariate Chi-square analyses, periodontitis and Alanine Aminotransferase were associated (p = 0.0360) and remained significant in unadjusted logistic regression (OR = 1.30 [95% CI: 1.02, 1.65]). However, when other known risk factors of periodontitis were included in the analyses, the relationship attenuated and failed to reach significance (adjusted OR = 1.17 [95% CI: 0.85, 1.60]). Conclusion. Our study adds to the literature a positive but attenuated association of serum Alanine Aminotransferase with periodontitis which failed to reach significance when other known, strong risk factors of periodontitis were included in the analysis.

Association of periodontitis and human papillomavirus in oral rinse specimens: Results from the National Health and Nutrition Survey 2009-2012.

BACKGROUND: Human papillomavirus (HPV)-positive oropharyngeal squamous cell carcinomas are increasing in incidence and are becoming significant public health concerns. Periodontitis is a chronic condition in which the affected tissue may facilitate oral HPV infection and persistence. The purpose of this study was to determine if an association of the presence of HPV in oral rinse specimens and periodontal disease exists. METHODS: The authors combined the National Health and Nutrition Examination Survey (NHANES) data for years 2009-2010 and 2011-2012. The authors included participants aged 30-69 years who had clinically assessed periodontal and HPV data (n = 6,004). The authors analyzed the data using the Rao-Scott χ(2) test and logistic regression. RESULTS: There were 498 participants who had the presence of HPV in oral rinse specimens. The adjusted odds ratio for the presence of HPV in oral rinse specimens with relation to periodontal disease was 1.04 (95% confidence interval, 0.63-1.73), adjusting for sex, race and ethnicity, education, age, income-to-poverty ratio, smoking, alcohol use, and number of sex partners during their lifetime. CONCLUSIONS: The authors failed to reject the hypothesis of no association of the presence of HPV in oral rinse specimens and periodontitis. PRACTICAL IMPLICATIONS: Although oral HPV infection is a serious concern, the authors found that periodontitis was not shown to be related to the presence of HPV in oral rinse specimens in adjusted analyses in this study.

A gold nanohole array based surface-enhanced Raman scattering biosensor for detection of silver(I) and mercury(II) in human saliva.

A surface-enhanced Raman scattering (SERS) biosensor has been developed by incorporating a gold nanohole array with a SERS probe (a gold nanostar@Raman-reporter@silica sandwich structure) into a single detection platform via DNA hybridization, which circumvents the nanoparticle aggregation and the inefficient Raman scattering issues. Strong plasmonic coupling between the Au nanostar and the Au nanohole array results in a large enhancement of the electromagnetic field, leading to amplification of the SERS signal. The SERS sensor has been used to detect Ag(I) and Hg(II) ions in human saliva because both the metal ions could be released from dental amalgam fillings. The developed SERS sensor can be adapted as a general detection platform for non-invasive measurements of a wide range of analytes such as metal ions, small molecules, DNA and proteins in body fluids.

Blood levels of the heavy metal, lead, and caries in children aged 24-72 months: NHANES III.

Lead remains a significant pollutant. It has acute toxic and chronic effects on many tissues and accumulates in teeth and bones. The researchers for this study investigated the association of blood lead levels with the extent/severity of caries as measured by the number of decayed/filled teeth of children aged 24-72 months using data from NHANES III (the Third National Health and Nutrition Examination Survey), accounting for the excess zero caries in the analysis and using less than 2 µg/dl as the reference blood lead level (n = 3,127). Zero-inflated negative binomial regression models indicated unadjusted extent/severity mean ratios of 1.79, 1.88 and 1.94 for the number of decayed/filled teeth in children whose blood lead levels were 2-5, 5-10 and >10 µg/dl, respectively, compared with children having <2 µg/dl blood lead levels. The results did not attenuate when other variables were added to the model for the 5-10 and >10 µg/dl levels of exposure. The adjusted extent/severity mean ratios were 1.84, 2.14 and 1.91, respectively, for the categories. This study indicated a strong association of blood lead levels with increasing numbers of carious teeth in children aged 24-72 months. These findings support other studies in an innovative analysis handling cases of children with no caries. The findings may inform caries risk assessment.

African ancestry influences CCR5 -2459G>A genotype-associated virologic success of highly active antiretroviral therapy.

INTRODUCTION: In a North American, HIV-positive, highly active antiretroviral therapy (HAART)-treated, adherent cohort of self-identified white and black patients, we previously observed that chemokine (C-C motif) receptor 5 (CCR5) -2459G>A genotype had a strong association with time to achieve virologic success (TVLS) in black but not in white patients. METHODS: Using 128 genome-wide ancestry informative markers, we performed a quantitative assessment of ancestry in these patients (n = 310) to determine (1) whether CCR5 -2459G>A genotype is still associated with TVLS of HAART when ancestry, not self-identified race, is considered and (2) whether this association is influenced by varying African ancestry. RESULTS: We found that the interaction between CCR5 -2459G>A genotype and African ancestry (≤ 0.125 vs. ≥ 0.425 and <0.71 vs. ≥ 0.71) was significantly associated with TVLS (GG compared with AA, P = 0.044 and 0.018, respectively). Furthermore, the association between CCR5 -2459G>A genotype and TVLS was stronger in patients with African ancestry ≥ 0.71 than in patients with African ancestry ≥ 0.452, in both Kaplan-Meier (log-rank P = 0.039 and 0.057, respectively, for AA, GA, and GG) and Cox proportional hazards regression (relative hazard for GG compared with AA 2.59 [95% confidence interval: 1.27 to 5.22; P = 0.01] and 2.26 [95% confidence interval: 1.18 to 4.32; P = 0.01], respectively) analyses. CONCLUSIONS: We observed that the association between CCR5 -2459G>A genotype and TVLS of HAART increased with stronger African ancestry. Understanding the genomic mechanisms by which African ancestry influences this association is critical and requires further studies.

Oral mycobiome analysis of HIV-infected patients: identification of Pichia as an antagonist of opportunistic fungi.

Oral microbiota contribute to health and disease, and their disruption may influence the course of oral diseases. Here, we used pyrosequencing to characterize the oral bacteriome and mycobiome of 12 HIV-infected patients and matched 12 uninfected controls. The number of bacterial and fungal genera in individuals ranged between 8-14 and 1-9, among uninfected and HIV-infected participants, respectively. The core oral bacteriome (COB) comprised 14 genera, of which 13 were common between the two groups. In contrast, the core oral mycobiome (COM) differed between HIV-infected and uninfected individuals, with Candida being the predominant fungus in both groups. Among Candida species, C. albicans was the most common (58% in uninfected and 83% in HIV-infected participants). Furthermore, 15 and 12 bacteria-fungi pairs were correlated significantly within uninfected and HIV-infected groups, respectively. Increase in Candida colonization was associated with a concomitant decrease in the abundance of Pichia, suggesting antagonism. We found that Pichia spent medium (PSM) inhibited growth of Candida, Aspergillus and Fusarium. Moreover, Pichia cells and PSM inhibited Candida biofilms (P = .002 and .02, respectively, compared to untreated controls). The mechanism by which Pichia inhibited Candida involved nutrient limitation, and modulation of growth and virulence factors. Finally, in an experimental murine model of oral candidiasis, we demonstrated that mice treated with PSM exhibited significantly lower infection score (P = .011) and fungal burden (P = .04) compared to untreated mice. Moreover, tongues of PSM-treated mice had few hyphae and intact epithelium, while vehicle- and nystatin-treated mice exhibited extensive fungal invasion of tissue with epithelial disruption. These results showed that PSM was efficacious against oral candidiasis in vitro and in vivo. The inhibitory activity of PSM was associated with secretory protein/s. Our findings provide the first evidence of interaction among members of the oral mycobiota, and identifies a potential novel antifungal.

Dental utilization of active duty/previous active duty US military: a cross-sectional analysis of the 2010 Behavior and Risk Surveillance Survey.

INTRODUCTION: The purpose of this study is to understand dental utilization of 1) individuals serving/having served in active duty in the U.S. military as compared with the general public and 2) individuals who are currently serving as compared with individuals who are no longer active duty, but have been in active duty within the previous year. METHODS: The Behavior and Risk Surveillance Survey, 2010, was used in cross-sectional analyses to determine the comparisons. Chi square and multivariable logistic regression analyses were applied. RESULTS: 70.7% of participants who had served/currently serving had a dental visit within the previous 12 months; 69.9% of the general public reported a dental visit (p = 0.0265). 69.8% of participants who had served/currently serving had a dental hygiene visit within the previous 12 months and 68.1% of the general public reported a dental hygiene visit (p <0.0001). The adjusted odds ratio (AOR) for participants who had served/currently serving vs. the general public was 1.10 (95% Confidence Interval [CI] 1.05, 1.16; p<0.0001) for dental visits and 1.11 (95%CI 1.05, 1.17; p<0.0001) for dental hygiene visits. CONCLUSION: Participants who are serving or have served were more likely to have any dental visit and dental hygiene visit than the general public; but the results were not substantively important.

Reduced systemic inflammatory mediators after treatment of chronic gingivitis.

OBJECTIVE: To assess the changes in the level of C-reactive protein (CRP) and tumor necrosis factor-alpha (TNF-alpha) in gingival crevicular fluid (GCF) after treatment of chronic gingivitis in systemically healthy individuals. METHODS: This is a clinical trial conducted at Case Western Reserve University, Cleveland, Ohio, United States of America from February to December 2011. A total of 41 systemically healthy subjects were assigned to 2 groups according to the severity of gingival inflammation. Group I consisted of 18 subjects who had mild gingival inflammation; and group II consisted of 23 with more severe gingival inflammation. Periodontal assessment consisted of gingival index (GI), probing depths (PD), and GCF volume. Four to six weeks after prophylaxis and oral hygiene instruction, the same measurements were repeated. The level of CRP and TNF-alpha in the GCF was determined using enzyme-linked immunosorbent assays. RESULTS: A statistically significant reduction in the mean CRP and TNF-alpha levels after the treatment was found in the severe, but not in the mild gingivitis group. Both groups showed a statistically significant reduction in GI, PD, and periotron readings after the treatment. CONCLUSION: Treatment of severe chronic gingivitis reduces the levels of CRP and TNF-alpha in GCF of otherwise systemically healthy individuals, which could have an impact on preventing or controlling future or existing systemic disease conditions.

Metabolomics reveals differential levels of oral metabolites in HIV-infected patients: toward novel diagnostic targets.

The objective of the current study was to characterize the profile of oral metabolites in HIV-infected patients using metabolomics. Oral wash samples were collected from 12 HIV-infected and 12 healthy individuals (matched for age, sex, and ethnicity), processed, and analyzed by metabolomics. We detected 198 identifiable and 85 nonidentifiable metabolites; 27 identifiable metabolites were differentially present (12 increased, 15 decreased) in HIV-infected patients. Elevated metabolites included p-cresol sulfate, nucleotides (e.g., allantoin), and amino acids (e.g., phenylalanine, tryptophan), whereas decreased oral metabolites included fucose, fumarate, and N-acetylglucosamine. Pathway network analysis revealed the largest multinode network in healthy versus HIV-infected patients to involve carbohydrate biosynthesis and degradation. HIV-infected patients on antiretroviral therapy (ART) showed the largest number (12) of statistically significant metabolite correlation differences compared with healthy controls. Interestingly, the oral phenlyalanine:tyrosine ratio increased in ART-naive HIV-infected patients (mean ± SEM = 2.58 ± 0.87) compared with healthy individuals (1.33 ± 0.10, p = 0.062) or ART-experienced patients (1.78 ± 0.30, p = 0.441). This is the first study to reveal differential levels of oral metabolites in HIV-infected patients compared withj healthy volunteers, and that oral phenlyalanine:tyrosine ratio may be a useful marker for noninvasive monitoring of the immune status during HIV infection.

Bacterial colonization and beta defensins in the female genital tract in HIV infection.

Beta defensins are antimicrobial peptides that serve to protect the host from microbial invasion at skin and mucosal surfaces. Here we explore the relationships among beta defensin levels, total bacterial colonization, and colonization by bacterial vaginosis (BV)-related bacteria and lactobacilli in the female genital tract in HIV infected women and healthy controls. Cervicovaginal lavage (CVL) samples were obtained from 30 HIV-infected women and 36 uninfected controls. Quantitative PCR assays were used to measure DNA levels of bacterial 16S ribosomal DNA (reflective of total bacterial load), and levels of three BV-related bacteria, three Lactobacillus species (L. crispatus, L. iners and L. jensenii), and total Lactobacillus levels in CVL. Levels of human beta defensins (hBD-2 and hBD-3) were quantified by ELISA. In viremic HIV+ donors, we found that CVL levels of bacterial 16S rDNA were significantly increased, and inversely correlated with peripheral CD4+ T cell counts in HIV+ women, and inversely correlated with age in both HIV+ women and controls. Although CVL DNA levels of BV-associated bacteria tended to be increased, and CVL levels of Lactobacillus DNAs tended to be decreased in HIV+ donors, none of these differences was significant. CVL levels of hBD-2 and hBD-3 were correlated and were not different in HIV+ women and controls. However, significant positive correlations between hBD-3 levels and total bacterial DNA levels in controls were not demonstrable in HIV+ women; the significant positive correlations of hBD2 or hBD-3 and three Lactobacillus species in controls were also not demonstrable in HIV+ women. These results suggest that HIV infection is associated with impaired regulation of innate defenses at mucosal sites.

Worldwide variation in human drug-metabolism enzyme genes CYP2B6 and UGT2B7: implications for HIV/AIDS treatment.

AIM: Hepatic enzymes, CYP2B6 and UGT2B7 play a major role in the metabolism of the widely used antiretroviral drugs efavirenz, nevirapine and zidovudine. In the present study, we provide a view of UGT2B7 haplotype structure, and quantify the genetic diversity and differentiation at both CYP2B6 and UGT2B7 genes on a worldwide scale. MATERIALS & METHODS: We genotyped one intronic and three promoter SNPs, and together with three nonsynonymous SNPs, inferred UGT2B7 alleles in north American (n = 326), west African (n = 133) and Papua New Guinean (n = 142) populations. We also included genotype data for five CYP2B6 and six UGT2B7 SNPs from an additional 12 worldwide populations (n = 629) analyzed in the 1000 Genomes Project. RESULTS: We observed significant differences in certain SNP and allele frequencies of CYP2B6 and UGT2B7 among worldwide populations. Diversity values were higher for UGT2B7 than for CYP2B6, although there was more diversity between populations for CYP2B6. For both genes, most of the genetic variation was observed among individuals within populations, with the Papua New Guinean population showing the highest pairwise differentiation values for CYP2B6, and the Asian and European populations showing higher pairwise differentiation values for UGT2B7. CONCLUSION: These new genetic distinctions provide additional insights for investigating differences in antiretroviral pharmacokinetics and therapy outcomes among ethnically and geographically diverse populations.

Copy Number Variation within Human ß-Defensin Gene Cluster Influences Progression to AIDS in the Multicenter AIDS Cohort Study.

STUDY BACKGROUND: DEFB4/103A encoding ß-defensin 2 and 3, respectively, inhibit CXCR4-tropic (X4) viruses in vitro. We determined whether DEFB4/103A Copy Number Variation (CNV) influences time-to-X4 and time-to-AIDS outcomes. METHODS: We utilized samples from a previously published Multicenter AIDS Cohort Study (MACS), which provides longitudinal account of viral tropism in relation to the full spectrum of rates of disease progression. Using traditional models for time-to-event analysis, we investigated association between DEFB4/103A CNV and the two outcomes, and interaction between DEFB4/103A CNV and disease progression groups, Fast and Slow. RESULTS: Time-to-X4 and time-to-AIDS were weakly correlated. There was a stronger relationship between these two outcomes for the fast progressors. DEFB4/103A CNV was associated with time-to-AIDS, but not time-to-X4. The association between higher DEFB4/103A CNV and time-to-AIDS was more pronounced for the slow progressors. CONCLUSION: DEFB4/103A CNV was associated with time-to-AIDS in a disease progression group-specific manner in the MACS cohort. Our findings may contribute to enhancing current understanding of how genetic predisposition influences AIDS progression.

Chemokine (C-C motif) receptor 5 -2459 genotype in patients receiving highly active antiretroviral therapy: race-specific influence on virologic success.

BACKGROUND: In patients receiving highly active antiretroviral therapy (HAART), antiretroviral drug-metabolizing enzyme and transporter gene polymorphisms, as well as chemokine receptor gene polymorphisms, may influence response to treatment. METHODS: In a North American, treated, adherent human immunodeficiency virus (HIV)-positive cohort (self-identified whites, n = 175; blacks, n = 218), we investigated whether CYP2B6 (516G>T, 983T>C), UGT2B7 (IVS1+985A>G, 802C>T), MDR1 3435C>T, chemokine (C-C motif) receptor 2 (CCR2) 190G>A, and CCR5 (-2459G>A, Δ32) polymorphisms influenced the time to achieve virologic success (TVLS). RESULTS: No difference in TVLS was observed between races. In Kaplan-Meier analyses, only 516G>T (log-rank P = .045 for comparison of GG, GT, and TT and P = .02 GG + GT vs TT) and -2459G>A (log-rank P = .04 for GG, GA, and AA and P = .02 for GG + GA vs AA) genotypes were significantly associated with TVLS in black patients but not in white patients. However, in the Cox proportional hazards model that included age, sex, baseline CD4(+) T cell count, and baseline viral load, no significant association was observed between 516G>T and TVLS, whereas the association between -2459G>A and TVLS remained significant even after including CCR2 190G>A as well as all the drug-metabolizing enzyme and transporter genotypes. CONCLUSIONS: These findings suggest that CCR5 -2459G>A genotype had a strong, race-specific influence on TVLS in this cohort. Understanding the possible mechanisms underlying this influence requires further studies.

Gentian violet exhibits activity against biofilms formed by oral Candida isolates obtained from HIV-infected patients.

The effect of gentian violet against Candida albicans and non-Candida albicans biofilms formed on polymethylmethacrylate strips was evaluated using a dry weight assay and confocal laser scanning microscopy. The ability of gentian violet to inhibit Candida albicans germination was also assessed. Gentian violet activity against Candida biofilms was demonstrated by a reduction in dry weight, disruption of biofilm architecture, and reduced biofilm thickness. Additionally, gentian violet inhibited Candida germination in a concentration-dependent manner.

Characterization of the oral fungal microbiome (mycobiome) in healthy individuals.

The oral microbiome-organisms residing in the oral cavity and their collective genome-are critical components of health and disease. The fungal component of the oral microbiota has not been characterized. In this study, we used a novel multitag pyrosequencing approach to characterize fungi present in the oral cavity of 20 healthy individuals, using the pan-fungal internal transcribed spacer (ITS) primers. Our results revealed the "basal" oral mycobiome profile of the enrolled individuals, and showed that across all the samples studied, the oral cavity contained 74 culturable and 11 non-culturable fungal genera. Among these genera, 39 were present in only one person, 16 genera were present in two participants, and 5 genera were present in three people, while 15 genera (including non-culturable organisms) were present in >/=4 (20%) participants. Candida species were the most frequent (isolated from 75% of participants), followed by Cladosporium (65%), Aureobasidium, Saccharomycetales (50% for both), Aspergillus (35%), Fusarium (30%), and Cryptococcus (20%). Four of these predominant genera are known to be pathogenic in humans. The low-abundance genera may represent environmental fungi present in the oral cavity and could simply be spores inhaled from the air or material ingested with food. Among the culturable genera, 61 were represented by one species each, while 13 genera comprised between 2 and 6 different species; the total number of species identified were 101. The number of species in the oral cavity of each individual ranged between 9 and 23. Principal component (PCO) analysis of the obtained data set followed by sample clustering and UniFrac analysis revealed that White males and Asian males clustered differently from each other, whereas both Asian and White females clustered together. This is the first study that identified the "basal mycobiome" of healthy individuals, and provides the basis for a detailed characterization of the oral mycobiome in health and disease.

Oral antimicrobial peptides and biological control of caries.

The presence of antimicrobial peptides (AMPs) in saliva may be a biological factor that contributes to susceptibility or resistance to caries. This manuscript will review AMPs in saliva, consider their antimicrobial and immunomodulatory functions, and evaluate their potential role in the oral cavity for protection of the tooth surface as well as the oral mucosa. These AMPs are made in salivary gland and duct cells and have broad antimicrobial activity. Alpha-defensins and LL37 are also released by neutrophils into the gingival crevicular fluid. Both sources may account for their presence in saliva. A recent study in middle school children aimed to determine a possible correlation between caries prevalence in children and salivary concentrations of the antimicrobial peptides human beta-defensin-3 (hBD-3), the cathelicidin, LL37, and the alpha-defensins. The levels of these AMPs were highly variable in the population. While levels of LL37 and hBD-3 did not correlate with caries experience, the mean alpha-defensin level was significantly higher in children with no caries than in children with caries (p < 0.005). We conclude that several types of AMPs that may have a role in oral health are present in unstimulated saliva. Low salivary levels of alpha-defensin may represent a biological factor that contributes to caries susceptibility. Our observation could lead to new ways to prevent caries and to a new tool for caries risk assessment.

Salivary antimicrobial peptide expression and dental caries experience in children.

Dental caries is a major worldwide oral disease problem in children. Although caries are known to be influenced by dietary factors, the disease results from a bacterial infection; thus, caries susceptibility may be affected by host factors such as salivary antimicrobial peptides. This study aimed to determine a possible correlation between caries prevalence in children and salivary concentrations of the antimicrobial peptides human beta-defensin-3 (hBD-3), the cathelicidin LL37, and the alpha-defensins HNP1-3 (a mixture of HNP1, 2, 3). Oral examinations were performed on 149 middle school children, and unstimulated whole saliva was collected for immunoassays of the three peptides and for assay of caries-causing bacteria in saliva. The median salivary levels of hBD-3, LL37, and HNP1-3 were in the microgram/ml range but were highly variable in the population. While levels of LL37 and hBD-3 did not correlate with caries experience, the median HNP1-3 levels were significantly higher in children with no caries than in children with caries. Children with high caries levels did not have high levels of salivary Streptococcus mutans, and the HNP1-3 level was not correlated with salivary S. mutans. By immunohistochemistry we localized HNP1-3 in submandibular salivary duct cells. HNPs are also released by neutrophils into the gingival crevicular fluid. Both sources may account for their presence in saliva. Low salivary levels of HNP1-3 may represent a biological factor that contributes to caries susceptibility. This observation could lead to new ways to screen for caries susceptibility and to new means of assessing the risk for this common oral problem.