Treated periodontitis and recurrent events after first-time myocardial infarction: A Danish nationwide cohort study.

AIM: To investigate the association between previous periodontal treatment and recurrent events after first-time myocardial infarction (MI). MATERIALS AND METHODS: From the Danish nationwide registries, patients with first-time MI between 2000 and 2015 were divided into three groups according to oral health care within 1 year prior to first-time MI. A multiple logistic regression model provided adjusted odds ratios (ORs) with 95% confidence intervals (CIs) to assess the 3-year risk of major adverse cardiovascular events (MACE). RESULTS: A total of 103,949 patients were included. Patients with treated periodontitis (PD) prior to first-time MI had an adjusted 3-year risk of MACE similar to patients presumed periodontally healthy (OR 0.97 [95% CI 0.92-1.03]). Patients with no prior dental visits were significantly older, had more comorbidities and showed significantly increased adjusted 3-year risks of MACE (OR 1.47 [95% CI 1.42-1.52]), cardiovascular death (OR 1.71 [95% CI 1.64-1.78]) and heart failure (OR 1.13 [95% CI 1.07-1.20]) compared with patients presumed periodontally healthy. CONCLUSIONS: Patients with treated PD 1 year prior to first-time MI had a similar risk of recurrent cardiovascular events as patients presumed periodontally healthy. No dental visit prior to first-time MI was an independent risk factor for recurrent events.

Additional information of bitewings to first time clinical examination of caries and restoration status in permanent dentition.

OBJECTIVES: Identifying additional information obtained by X-rays combined with clinical examination concerning primary caries, caries adjacent to restorations and quality of restorations. MATERIAL AND METHODS: A total of 240 adult patients, equally distributed in gender and six age-groups, were randomly selected from an original study population of 4,402 subjects (DANHES). Clinical and radiographical registrations on occlusal and approximal surfaces in posterior teeth were categorized into unrestored surfaces (sound/primary caries) and restored surfaces (without/with caries adjacent to restorations). Material and quality of restorations were also recorded. Chi-square and Fisher-exact tests were used for statistical analyses. RESULTS: Of potentially 11,520 surfaces, 3,015 occlusal and 5,112 approximal surfaces were analysed. Occlusal: Of 907 unrestored surfaces, 110 had primary caries and 53% were detected radiographically. A total of 183 of 2,108 restored surfaces had caries adjacent to restorations, and 99% were found radiographically. A total of 190 restorations were over/under-extended, and 89% were registered radiographically. Approximal: Of 2,649 unrestored surfaces 648 had primary caries, and 92% were registered radiographically. A total of 565 of 2,463 restored surfaces had caries adjacent to restorations, and 99% were found radiographically. A total of 638 restorations were over/under-extended, and 98% were found radiographically. For all restorations, material and depth influenced quality of restorations and incidence of caries adjacent to restorations. At least one independent variable (gender/age group/tooth-type/jaw/side/mesial-distal surface) influenced frequencies of primary caries, caries adjacent to restorations and quality of restorations. CONCLUSIONS: First-time clinical examination must be supplemented with X-rays to obtain a complete impression of caries status in posterior regions regarding diagnostics of caries, assessment of lesion depth and quality of restorations.

Immunoglobulin G antibodies against Porphyromonas gingivalis or Aggregatibacter actinomycetemcomitans in cardiovascular disease and periodontitis.

Objectives: The aim was to elucidate whether levels of circulating antibodies to Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis correlate to loss of attachment, as a marker for periodontitis and cardiovascular disease (CVD). Design: Sera were collected from 576 participants of the Danish Health Examination Survey (DANHES). Immunoglobulin G antibodies against lipopolysaccharide (LPS) and protein antigens from the a, b and c serotypes of A. actinomycetemcomitans and P. gingivalis were quantified by titration in ELISA plates coated with a mixture of antigens prepared by disintegration of bacteria. Results: Levels of antibodies against P. gingivalis (OR = 1.48) and A. actinomycetemcomitans (1.31) associated with periodontitis, as determined by univariable logistic regression analysis. These antibody levels also associated with CVD (1.17 and 1.37), respectively, However, after adjusting for other risk factors, including age, smoking, gender, alcohol consumption, overweight, and level of education using multivariable logistic regression analysis, only increasing body mass index (BMI; 1.09), previous smoking (1.99), and increasing age (decades) (2.27) remained associated with CVD. Increased levels of antibodies against P. gingivalis (1.34) remained associated with periodontitis after adjusting for other risk factors. Conclusions: CVD and periodontitis were associated with levels of IgG antibodies to P. gingivalis or A. actinomycetemcomitans in univariable analyses, but only the association of P. gingivalis antibody levels with periodontitis reached statistical significance after adjustment for common confounders. Age, in particular, influenced this relationship.

Dental biofilm infections - an update.

Teeth are colonized by oral bacteria from saliva containing more than 700 different bacterial species. If removed regularly, the dental biofilm mainly comprises oral streptococci and is regarded as resident microflora. But if left undisturbed, a complex biofilm containing up to 100 bacterial species at a site will build up and may eventually cause development of disease. Depending on local ecological factors, the composition of the dental biofilm may vary considerably. With access to excess carbohydrates, the dental biofilm will be dominated by mainly gram-positive carbohydrate-fermenting bacteria causing demineralization of teeth, dental caries, which may further lead to inflammation and necrosis in the pulp and periapical region, i.e., pulpitis and periapical periodontitis. In supra- and subgingival biofilms, predominantly gram-negative, anaerobic proteolytic bacteria will colonize and cause gingival inflammation and breakdown of supporting periodontal fibers and bone and ultimately tooth loss, i.e., gingivitis, chronic or aggressive periodontitis, and around dental implants, peri-implantitis. Furthermore, bacteria from the dental biofilm may spread to other parts of the body by bacteremia and cause systemic disease. Basically, prevention and treatment of dental biofilm infections are achieved by regular personal and professional removal of the dental biofilm.

Salivary microbiota in individuals with different levels of caries experience.

This study compared salivary bacterial profiles in two groups having a 10-fold difference in levels of caries experience, as it was hypothesized that the composition of the salivary microbiota might associate with the levels of caries experience. Bacterial profiles in stimulated saliva samples from 85 individuals with low levels of caries experience (healthy group) and 79 individuals with high levels of caries experience (caries group) were analyzed by means of the Human Oral Microbiome Identification Next Generation Sequencing (HOMINGS) technique. Subsequently, saliva samples from caries-free individuals in the healthy group (n = 57) and the caries group (n = 31) were compared. A significantly higher α-diversity (p < 0.0001) and a twofold higher relative abundance of Neisseria, Haemophilus, and Fusobacterium were recorded in saliva samples from the healthy group compared with the caries group. Differences observed were more pronounced when limiting the analyses to caries-free individuals in each group. Data from this cross-sectional analysis suggest that low levels of caries experience might associate with a characteristic salivary bacterial composition different from that in individuals with high caries experience. Consequently, longitudinal studies are required to determine if the composition of the salivary microbiota might be a predictive factor of caries risk at the individual level.

Impact of Periodontitis Case Criteria: A Cross-Sectional Study of Lifestyle.

BACKGROUND: Associations of risk factors/indicators with periodontitis may depend on the included case criterion. The objective of the current study is to evaluate differences in outcome by applying five periodontitis case definitions for cross-sectional associations with lifestyle factors among participants of the Danish Health Examination Survey (DANHES). METHODS: A total of 4,402 adults aged 18 to 96 years from the general health examination of DANHES had a periodontal examination consisting of half-mouth registration at six sites per tooth including probing depth (PD) and clinical attachment level (CAL). Periodontitis was defined according to severe periodontitis, European Workshop of Periodontology (EWP)-specific, meanCAL ≥2.55 mm, CAL-tertile, and PD-CAL definitions. Multivariable logistic regression models fitted the association of age, sex, smoking status, diabetes mellitus, educational level, alcohol consumption, body mass index, physical activity, body fat percentage, waist circumference, triglycerides, total cholesterol, and C-reactive protein with periodontitis. RESULTS: Number of cases captured by the five periodontitis case definitions ranged from 337 (9.2%) to 1,136 (31.0%). A total of 224 participants were defined as periodontitis cases by all five criteria. Analyses on 3,665 participants with complete data revealed statistically significant associations of age and smoking with all periodontitis case definitions and of male sex with severe periodontitis and EWP-specific definitions. Educational level (two lowest groups) was related to three periodontitis criteria. Among obesity and hyperlipidemia measures no factors were related to periodontitis. CONCLUSION: Regression analyses showed little difference in odds ratio across the five periodontitis case definitions; however, the level of significance did show some variation.

Comparative analysis of bacterial profiles in unstimulated and stimulated saliva samples.

BACKGROUND AND OBJECTIVE: The microbial profiles of stimulated saliva samples have been shown to differentiate between patients with periodontitis, patients with dental caries, and orally healthy individuals. Saliva was stimulated to allow for easy and rapid collection; however, microbial composition may not reflect the more natural, unstimulated state. The purpose of this study was to validate whether stimulated saliva is an adequate surrogate for unstimulated saliva in determining salivary microbiomes. DESIGN: Unstimulated (n=20) and stimulated (n=20) saliva samples were collected from 20 orally and systemically healthy, non-smoking participants. Salivary bacterial profiles were analyzed by means of the Human Oral Microbe Identification using Next Generation Sequencing (HOMINGS), and statistical analysis was performed using Mann-Whitney test with Benjamini-Hochberg's correction for multiple comparison, cluster analysis, principal component analysis, and correspondence analysis. RESULTS: From a total of 40 saliva samples, 496 probe targets were identified with a mean number of targets per sample of 203 (range: 146-303), and a mean number of probe targets of 206 and 200 in unstimulated and stimulated saliva samples, respectively (p=0.62). Based on all statistical methods used for this study, the microbial profiles of unstimulated and stimulated saliva samples collected from the same person were not statistically significantly different. CONCLUSIONS: Analysis of bacterial salivary profiles in unstimulated and stimulated saliva samples collected from the same individual showed comparable results. Thus, the results verify that stimulated saliva is an adequate surrogate of unstimulated saliva for microbiome-related studies.

Salivary bacterial fingerprints of established oral disease revealed by the Human Oral Microbe Identification using Next Generation Sequencing (HOMINGS) technique.

BACKGROUND AND OBJECTIVE: The composition of the salivary microbiota, as determined using various molecular methods, has been reported to differentiate oral health from diseases. Thus, the purpose of this study was to utilize the newly developed molecular technique HOMINGS (Human Oral Microbe Identification using Next Generation Sequencing) for comparison of the salivary microbiota in patients with periodontitis, patients with dental caries, and orally healthy individuals. The hypothesis was that this method could add on to the existing knowledge on salivary bacterial profiles in oral health and disease. DESIGN: Stimulated saliva samples (n=30) were collected from 10 patients with untreated periodontitis, 10 patients with untreated dental caries, and 10 orally healthy individuals. Salivary microbiota was analyzed using HOMINGS and statistical analysis was performed using Kruskal-Wallis test with Benjamini-Hochberg's correction. RESULTS: From a total of 30 saliva samples, a mean number of probe targets of 205 (range 120-353) were identified, and a statistically significant higher mean number of targets was registered in samples from patients with periodontitis (mean 220, range 143-306) and dental caries (mean 221, range 165-353) as compared to orally healthy individuals (mean 174, range 120-260) (p=0.04 and p=0.04). Nine probe targets were identified with a different relative abundance between groups (p<0.05). CONCLUSIONS: Cross-sectional comparison of salivary bacterial profiles by means of HOMINGS analysis showed that different salivary bacterial profiles were associated with oral health and disease. Future large-scale prospective studies are needed to evaluate if saliva-based screening for disease-associated oral bacterial profiles may be used for identification of patients at risk of acquiring periodontitis and dental caries.

Temporal Stability of the Salivary Microbiota in Oral Health.

OBJECTIVES: Saliva is a biological fluid suitable for biomarker analysis, and differences in the salivary microbiota in oral health and disease have been reported. For such comparative analyses, time of sampling is critical since the bacterial composition may vary throughout the day, i.e., diurnal variation. The purpose of this study is to compare the salivary microbiome over time to determine the optimal time for sampling. DESIGN: Stimulated saliva samples were collected from 5 orally healthy individuals in 4 h intervals for 24 h, and collection was repeated 7 days later (number of samples per person, n = 12, total number of samples, n = 60). Salivary microbiota was analyzed using the Human Oral Microbe Identification using Next Generation Sequencing (HOMINGS), and statistical analysis was performed using the Kruskal-Wallis test with Benjamini-Hochberg's correction for multiple comparisons, cluster analysis, principal component analysis and correspondence analysis. RESULTS: From a total of 60 saliva samples, 477 probe targets were collectively identified with a mean number of probes per sample of 207 (range: 153-307). Little or no variation in microbial profiles within subjects was observed over time. CONCLUSIONS: Although there was considerable variation between subjects, microbial profiles within subjects were stable throughout a 24 hour period and after 1 week. Since there is little or no evidence of diurnal variation of the salivary microbiome, time of sampling of saliva is not critical for perturbation or other microbial studies.

Calcium, vitamin D, casein and whey protein intakes and periodontitis among Danish adults.

OBJECTIVE: To investigate whether intakes of Ca, vitamin D, casein and whey are associated with periodontitis and to investigate the possibility of interactions between them. DESIGN: Cross-sectional study. An Internet-based, 267-item FFQ was used to assess dietary intake. Intakes of casein (32.0 g/d), whey proteins (9.6 g/d) and vitamin D (5.8 µg/d) were classified as within v. above the 50th percentile. Ca intake was classified as within v. below age-specific recommendations. Severe periodontitis was defined as having ≥2 inter-proximal sites with clinical attachment loss ≥6 mm (not on the same tooth) and ≥1 inter-proximal site with pocket depth ≥5 mm. Since vitamin D influences Ca absorption, models were stratified by lower and higher (<5.8 v. ≥5.8 µg/d) vitamin D intake. SETTING: Danish Health Examination Survey (DANHES) 2007-2008. SUBJECTS: Adult participants (n 3287) in the oral health study of DANHES 2007-2008. RESULTS: Intakes of Ca within recommendations (OR=0.76; 95% CI 0.58, 0.99), whey ≥9.6 g/d (OR=0.75; 95% CI 0.58, 0.97) and casein ≥32 g/d (OR=0.75 95% CI 0.58, 0.97) were associated with lower likelihood of severe periodontitis after adjustment for age, gender, education, smoking, sucrose intake, alcohol consumption, number of teeth, daily brushing, regular visits to the dentist and chronic illness, irrespective of vitamin D intake levels. Intake of vitamin D alone was not associated severe with periodontitis. CONCLUSIONS: Intakes of Ca, casein and whey protein were inversely associated with periodontitis. Consumption of foods rich in Ca, casein and whey (e.g. dairy foods) should be promoted, as they may contribute to the prevention of periodontitis. Further longitudinal studies are required to confirm these associations.

Differentiation of salivary bacterial profiles of subjects with periodontitis and dental caries.

Bacterial profiles of saliva in subjects with periodontitis and dental caries have been demonstrated to differ from that of oral health. The aim of this comparative analysis of existing data generated by the Human Oral Microbe Identification Microarray (HOMIM) from 293 stimulated saliva samples was to compare bacterial profiles of saliva in subjects with periodontitis and dental caries.

Root caries, root surface restorations and lifestyle factors in adult Danes.

AIM: To investigate selected lifestyle factors in relation to active caries and restored root surface lesions in adults. MATERIALS AND METHODS: Based on clinical examinations and questionnaires, data on root caries, socioeconomic status, body mass index, dietary habits, alcohol consumption, tobacco use and oral hygiene routines were collected from 4369 adults aged 21-89 who took part in a survey covering 13 municipalities across Denmark. Uni- and multivariate logistic regression analyses were applied to analyse the relationship between the independent lifestyle variables and active caries and restored root surface lesions, respectively. RESULTS: The prevalence of active root caries was 4%, while 26% displayed restored root surfaces. The sugar intake was not related to root caries. A multivariate logistic regression analysis revealed that, in subjects aged 45 or over, smoking and wearing dentures were significantly associated with presence of active root caries (p<0.01). The intake of 15 drinks or more per week was associated with higher odds of root surface restorations compared with no alcohol intake (OR=1.7; p<0.001). CONCLUSIONS: Lifestyle factors such as tobacco use and alcohol consumption, as well as wearing dentures, were significantly associated with the occurrence of untreated caries and restored root surface lesions, especially in persons over 45. Thus, such lifestyle factors should be taken into consideration, identifying persons with a need of preventive dental services. In addition, oral health education should focus on the possible risks of smoking and a high alcohol intake.

Bacterial profiles of saliva in relation to diet, lifestyle factors, and socioeconomic status.

BACKGROUND AND OBJECTIVE: The bacterial profile of saliva is composed of bacteria from different oral surfaces. The objective of this study was to determine whether different diet intake, lifestyle, or socioeconomic status is associated with characteristic bacterial saliva profiles. DESIGN: Stimulated saliva samples from 292 participants with low levels of dental caries and periodontitis, enrolled in the Danish Health Examination Survey (DANHES), were analyzed for the presence of approximately 300 bacterial species by means of the Human Oral Microbe Identification Microarray (HOMIM). Using presence and levels (mean HOMIM-value) of bacterial probes as endpoints, the influence of diet intake, lifestyle, and socioeconomic status on the bacterial saliva profile was analyzed by Mann-Whitney tests with Benjamini-Hochberg's correction for multiple comparisons and principal component analysis. RESULTS: Targets for 131 different probes were identified in 292 samples, with Streptococcus and Veillonella being the most predominant genera identified. Two bacterial taxa (Streptococcus sobrinus and Eubacterium [11][G-3] brachy) were more associated with smokers than non-smokers (adjusted p-value<0.01). Stratification of the group based on extreme ends of the parameters age, gender, alcohol consumption, body mass index (BMI), and diet intake had no statistical influence on the composition of the bacterial profile of saliva. Conversely, differences in socioeconomic status were reflected by the bacterial profiles of saliva. CONCLUSIONS: The bacterial profile of saliva seems independent of diet intake, but influenced by smoking and maybe socioeconomic status.

Smoking and drinking as risk indicators for tooth loss in middle-aged Danes.

OBJECTIVES: To investigate tobacco and alcohol consumption as risk indicators for missing teeth in late middle-aged Danes. METHOD: In all, 1,517 Copenhagen Aging and Midlife Biobank (CAMB) participants received a clinical oral examination that included number of teeth. Information on smoking, drinking, and various covariates was obtained using self-administered, structured questionnaires. Descriptive statistics and logistic regression (dependent variable: 6+ vs. <6 missing teeth) were used to investigate smoking and drinking in relation to missing teeth. RESULTS: Current smokers, persons who currently or previously smoked >15 tobacco units/day, and persons who had smoked for 27+ years had elevated mean scores of missing teeth and associated odds ratios (OR) compared with never smokers. Relative to nondrinkers, alcohol consumption was associated with reduced odds of missing 6+ teeth. DISCUSSION: Our findings suggest that smoking is positively associated, while alcoholic beverage consumption is inversely related to tooth loss in middle-aged Danes.

Differences in bacterial saliva profile between periodontitis patients and a control cohort.

AIM: Periodontitis is a multifactorial disease in which subgingival bacteria play an important role in the pathogenesis of the disease. The objective of this study was to determine if periodontitis is associated with a characteristic salivary bacterial profile. This was accomplished by comparing the bacterial profile of saliva from subjects with chronic periodontitis with that of saliva from a control cohort. MATERIALS AND METHODS: Stimulated saliva samples from 139 chronic periodontitis patients and 447 samples from a control cohort were analysed using the Human Oral Microbe Identification Microarray (HOMIM). Frequency and levels (mean HOMIM-value) of around 300 bacterial taxa/clusters in samples were used as parameters for investigation. Differences at taxon/cluster values between groups were analysed using Mann-Whitney U-test with Benjamini-Hochberg correction for multiple comparisons. Principal component analysis was used to visualize bacterial community profiles obtained by the HOMIM. RESULTS: Eight bacterial taxa, including putative periodontal pathogens as Parvimonas micra and Filifactor alocis, and four bacterial clusters were identified statistically more frequently and at higher levels in samples from periodontitis patients than in samples from the control cohort. These differences were independent of the individuals' smoking status. CONCLUSIONS: Periodontitis is associated with a characteristic bacterial profile of saliva different from that of a control cohort.

Bacterial sex in dental plaque.

Genes are transferred between bacteria in dental plaque by transduction, conjugation, and transformation. Membrane vesicles can also provide a mechanism for horizontal gene transfer. DNA transfer is considered bacterial sex, but the transfer is not parallel to processes that we associate with sex in higher organisms. Several examples of bacterial gene transfer in the oral cavity are given in this review. How frequently this occurs in dental plaque is not clear, but evidence suggests that it affects a number of the major genera present. It has been estimated that new sequences in genomes established through horizontal gene transfer can constitute up to 30% of bacterial genomes. Gene transfer can be both inter- and intrageneric, and it can also affect transient organisms. The transferred DNA can be integrated or recombined in the recipient's chromosome or remain as an extrachromosomal inheritable element. This can make dental plaque a reservoir for antimicrobial resistance genes. The ability to transfer DNA is important for bacteria, making them better adapted to the harsh environment of the human mouth, and promoting their survival, virulence, and pathogenicity.

Findings from the oral health study of the Danish Health Examination Survey 2007-2008.

OBJECTIVE: The aims of the oral part of the Danish Health Examination Survey (DANHES 2007-2008) were (1) to establish an oral health database for adult Danes and (2) to explore the influence of general diseases and lifestyle on oral health. This paper presents the study population, examination methods, questionnaire and baseline results. MATERIALS AND METHODS: The study population comprised 4402 subjects, aged 18-96, consecutively enrolled from 18 065 DANHES participants from 13 municipalities in Denmark. The oral part consisted of a validated questionnaire and a clinical examination, carried out in mobile units by three trained and calibrated dental hygienists. The data were processed with descriptive statistics and mono- and bivariate analyses. RESULTS: The mean age was 54.1 years and 60% were women. The mean number of natural teeth was 26.6; the mean DMFT/DMFS values were 18.9 and 61.0, and varied with age (DMFT 8.7-24.3). A higher proportion of females suffered from dental erosion in the younger age groups. Forty per cent of all subjects had a mean clinical attachment loss ≥ 3 mm, varying from 4% among those aged 18-34 to 80% in those over 75. A sub-optimal saliva secretion rate was more common among females than males (17.7% vs 10.4%) and this was reflected by the reported frequency of dry mouth. CONCLUSION: This extensive cross-sectional study provides a platform for obtaining future knowledge of the impact of health- and lifestyle-related factors on oral diseases. The validated questionnaire and the clinical characteristics enable robust analyses, although the conclusions may be hampered by limited external validity.