Walter A. Bretz

Periodontal Disease and the Oral Microbiota in New-Onset Rheumatoid Arthritis

OBJECTIVE To profile the abundance and diversity of subgingival oral microbiota in patients with never-treated, new-onset rheumatoid arthritis (RA). METHODS Periodontal disease (PD) status, clinical activity, and sociodemographic factors were determined in patients with new-onset RA, patients with chronic RA, and healthy subjects. Multiplexed-454 pyrosequencing was used to compare the composition of subgingival microbiota and establish correlations between the presence/abundance of bacteria and disease phenotypes. Anti-Porphyromonas gingivalis antibody testing was performed to assess prior exposure to the bacterial pathogen P gingivalis. RESULTS The more advanced forms of periodontitis were already present at disease onset in patients with new-onset RA. The subgingival microbiota observed in patients with new-onset RA was distinct from that found in healthy controls. In most cases, however, these microbial differences could be attributed to the severity of PD and were not inherent to RA. The presence and abundance of P gingivalis were also directly associated with the severity of PD and were not unique to RA. The presence of P gingivalis was not correlated with anti-citrullinated protein antibody (ACPA) titers. Overall exposure to P gingivalis was similar between patients with new-onset RA and controls, observed in 78% of patients and 83% of controls. The presence and abundance of Anaeroglobus geminatus correlated with the presence of ACPAs/rheumatoid factor. Prevotella and Leptotrichia species were the only characteristic taxa observed in patients with new-onset RA irrespective of PD status. CONCLUSION Patients with new-onset RA exhibited a high prevalence of PD at disease onset, despite their young age and paucity of smoking history. The subgingival microbiota profile in patients with new-onset RA was similar to that in patients with chronic RA and healthy subjects whose PD was of comparable severity. Although colonization with P gingivalis correlated with the severity of PD, overall exposure to P gingivalis was similar among the groups. The role of A geminatus and Prevotella/Leptotrichia species in this process merits further study.

Microbial Risk Indicators of Early Childhood Caries

ABSTRACT The aim of this study was to use molecular identification methods, such as 16S RNA gene sequence and reverse-capture checkerboard hybridization, for identification of the bacteria associated with dental caries and with dental health in a subset of 204 twins aged 1.5 to 7 years old. A total of 448 plaque samples (118 collected from caries-free subjects and 330 from caries-active subjects) were used for analysis. We compared the bacteria found in biofilms of children exhibiting severe dental caries, with different degrees of lesion severity, with those found in biofilms of caries-free children. A panel of 82 bacterial species was selected, and a PCR-based reverse-capture checkerboard method was used for detection. A simple univariate test was used to determine the overabundance and underabundance of bacterial species in the diseased and in the healthy groups. Features identified with this univariate test were used to construct a probabilistic disease prediction model. Furthermore, a method for the analysis of global patterns of gene expression was performed to permit simultaneous analysis of the abundance of significant species by allowing cross-bacterial comparisons of abundance profiles between caries-active and caries-free subjects. Our results suggested that global patterns of microbial abundance in this population are very distinctive. The top bacterial species found to be overabundant in the caries-active group were Actinomyces sp. strain B19SC, Streptococcus mutans, and Lactobacillus spp., which exhibited an inverse relationship to beneficial bacterial species, such as Streptococcus parasanguinis, Abiotrophia defectiva, Streptococcus mitis, Streptococcus oralis, and Streptococcus sanguinis.

Anti-inflammatory effects of a bioavailable compound, Artepillin C, in Brazilian propolis

Artepillin C is the major compound in the Brazilian green propolis from Baccharis dracunculifolia. Our aim in this study was to investigate the anti-inflammatory effects, absorption, and bioavailability of Artepillin C in mice. The animals used were male Swiss mice subjected to: paw oedema by carrageenan (300 microg/paw), carrageenan-induced peritonitis, and prostaglandin E(2) determination. We also measured in vitro nitric oxide production by RAW 264.7 cells and NF-kappaB activity in HEK 293 cells. Finally, we measured the absorption and bioavailability of Artepillin C in plasma from mice by means of GC-MS after a single oral dose (10 mg/kg). In vivo, Artepillin C produced a maximal inhibition of 38% after 360 min on paw oedema. Artepillin C also decreased the number of neutrophils during peritonitis (IC(50): 0.9 (0.5-1.4) mg/kg). Treatment with Artepillin C decreased prostaglandin E(2) by 29+/-3% and 58+/-5% at 1 and 10 mg/kg, respectively, with a mean ID(50) of 8.5 (8.0-8.7) mg/kg). Similarly, in in vitro models, Artepillin C (3, 10, or 100 microM) decreased nitric oxide production by RAW 264.7 cells with a mean IC(50) of 8.5 (7.8-9.2) microM. In HEK 293 cells, Artepillin C reduced NF-kappaB activity with a mean IC(50) of 26 (22-30) mug/ml), suggesting anti-inflammatory activity, particularly during acute inflammation. Lastly, Artepillin C was absorbed after an oral dose (10 mg/kg) with maximal peaks found at 1 h (22 microg/ml). Collectively, Artepillin C showed anti-inflammatory effects mediated, at least in part, by prostaglandin E(2) and nitric oxide inhibition through NF-kappaB modulation, and exhibited bioavailability by oral administration.

Xerostomia, Xerogenic Medications and Food Avoidances in Selected Geriatric Groups

OBJECTIVE: To study the relationship between complaints of xerostomia and salivary performance and food avoidances in four geriatric groups chosen to reflect a broad spectrum of individuals along the health‐disease continuum. To determine whether xerogenic medications taken by these individuals could be associated with either complaints of xerostomia or with food avoidances.

Antibacterial activity of honey and propolis from Apis mellifera and Tetragonisca angustula against Staphylococcus aureus

Aims: The antibacterial activity against Staphylococcus aureus of honey and propolis produced by Apis mellifera and Tetragonisca angustula was evaluated. Secondary aims included the study of the chemical composition of propolis and honey samples and its relationship with antibacterial activity against S. aureus.

Longitudinal Analysis of Heritability for Dental Caries Traits

The role of genetic and environmental factors on dental caries progression in young children was determined. A detailed caries assessment was performed in 2 examinations on 314 pairs of twins initially 1.5 to 8 years old. Surface-based caries prevalence rates (SBCPR) and lesion severity (LSI) were computed. Heritability estimates were calculated by SOLAR software. Analyses were performed on all ages combined and by age group (1.5-< 4; 4–6; > 6). Overall heritability estimates (H) of net increments SBCPRs were H = 30.0 (p < 0.0001), and were greatest for the youngest (H = 30.0) and oldest groups (H = 46.3). Overall LSI heritability estimates [H = 36.1 (p < 0.0001)] were also greatest for the youngest (H = 51.2) and oldest groups (H = 50.6). Similar findings were found for net increments of occlusal surfaces and deep dentinal lesions SBCPRs (H = 46.4–56.2). These findings are consistent with a significant genetic contribution to dental caries progression and severity in both emerging primary and permanent dentitions.

Effect of Propolis on Human Fibroblasts from the Pulp and Periodontal Ligament

Propolis, a flavonoid-rich product of honey comb, exhibits antibacterial and anti-inflammatory properties. In this study, we examined the tolerance of fibroblasts of the periodontal ligament (PDL) and dental pulp to propolis and compared with that of calcium hydroxide in vitro. Cells from human dental pulp and PDL were obtained from healthy third molars and subjected to various concentrations of propolis (0-20 mg/ml) and calcium hydroxide (0-250 mg/ml). The cell viability after propolis treatment was analyzed by crystal violet staining of the cells followed by spectrophotometric analysis. Data revealed that exposure of PDL cells or pulp fibroblasts to 4 mg/ml or lower concentrations of propolis resulted in >75% viability of cells. On the contrary, calcium hydroxide 0.4 mg/ml was cytotoxic and <25% of the cells were found to be viable. Further investigations may find propolis to be a possible alternative for an intracanal antimicrobial agent.

The Dental Plaque Microbiome in Health and Disease

Dental decay is one of the most prevalent chronic diseases worldwide. A variety of factors, including microbial, genetic, immunological, behavioral and environmental, interact to contribute to dental caries onset and development. Previous studies focused on the microbial basis for dental caries have identified species associated with both dental health and disease. The purpose of the current study was to improve our knowledge of the microbial species involved in dental caries and health by performing a comprehensive 16S rDNA profiling of the dental plaque microbiome of both caries-free and caries-active subjects. Analysis of over 50,000 nearly full-length 16S rDNA clones allowed the identification of 1,372 operational taxonomic units (OTUs) in the dental plaque microbiome. Approximately half of the OTUs were common to both caries-free and caries-active microbiomes and present at similar abundance. The majority of differences in OTU’s reflected very low abundance phylotypes. This survey allowed us to define the population structure of the dental plaque microbiome and to identify the microbial signatures associated with dental health and disease. The deep profiling of dental plaque allowed the identification of 87 phylotypes that are over-represented in either caries-free or caries-active subjects. Among these signatures, those associated with dental health outnumbered those associated with dental caries by nearly two-fold. A comparison of this data to other published studies indicate significant heterogeneity in study outcomes and suggest that novel approaches may be required to further define the signatures of dental caries onset and progression.

Dental Caries and Microbial Acid Production in Twins

Objective: To determine the relative contribution of genetic and environmental stimuli on dental caries traits and microbial acid production in a twin model. Methods: Dental caries examinations and microbial acid production assays were performed on 388 pairs of twins 1.5–8 years old from the city of Montes Claros, Brazil. Genotyping 8 polymorphic DNA markers determined zygosity. Caries exams followed NIDCR criteria modified to distinguish white spot lesions from cavitated lesions. Surface-based caries prevalence rates (SBCPR) were computed and lesion severity was determined by a weighted index (LSI). Biofilm samples were collected from the tongue using a lactic acid indicator swab. Assay scores were categorized based on acid formation as 1 = low, 2 = medium, and 3 = high. Heritability analyses were performed using the SOLAR software package. Results: Heritability estimates for SBCPRs, LSI and for microbial acid production were H = 76.3 (p < 0.001), H = 70.6 (p < 0.001), H = 16.2 (p = 0.0078), respectively. Treating microbial acid production as a covariate in the SBCPR and LSI models did not significantly alter the heritability estimates, i.e. H = 76.5 (p < 0.001) and H = 70.8 (p < 0.001), respectively. Conclusions: These results suggest that variation in dental caries surface traits has a significant genetic contribution and that microbial acid production is modulated by the environment.

Periodontitis and Airway Obstruction

BACKGROUND The objective of this study was to examine the relationship between airway obstruction and periodontal disease. METHODS Participants were a subset of 860 community- dwelling, well functioning elderly (aged 70 to 79, blacks and whites, males and females) selected from 2,732 participants enrolled in the Health, Aging, and Body Composition Study (Health ABC). The periodontal evaluations occurred over years 2 and 3 of the study and included four indices of periodontal health: plaque index (PI), gingival index (GI), probing depth (PD), and loss of attachment (LOA). The pulmonary evaluation took place in year 1: conducted according to American Thoracic Society criteria, based on the forced expiratory volume/forced vital capacity (FEV1 /FVC) ratio and then using the percent of predicted FEV1 to categorize severity. RESULTS GI (P = 0.023) and LOA (P = 0.009) were significantly better in participants with normal pulmonary function compared to those with airway obstruction after adjusting for age, race, gender, and field center. When stratified by smoking status and after adjusting for age, race, gender, center, and pack-years, there was a significant association between periodontal health and airway obstruction in former smokers. Within this group, those with normal pulmonary function had significantly better GI (P = 0.036) and LOA (P = 0.0003) scores than those with airway obstruction. All periodontal indices were elevated in smokers regardless of pulmonary status; however, the current smoker group was too small to detect a periodontitis effect. CONCLUSION While the present cross-sectional study cannot provide direct inference of cause and effect, it does reveal a significant association between periodontal disease and airway obstruction, particularly in former smokers.