van Arie Winkelhoff

Polymorphisms of the Interleukin-1 Gene Family, Oral Microbial Pathogens, and Smoking in Adult Periodontitis

Interleukin (IL)-1α, IL-1β, and lL-1ra contribute to regulation of the inflammatory response in periodontal tissues. We aimed to investigate the distribution of polymorphisms in the IL-1 gene family among periodontitis patients and controls, taking into account smoking and microbiology as additional variables. Fifty-three non-smoking and 52 smoking patients with severe adult periodontitis and 53 controls were genotyped for bi-allelic IL-1A-889, IL-1B +3954 , and a penta-allelic 86-bp VNTR IL-1RN gene polymorphisms. The presence of Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans was established by culture techniques. We found a higher frequency of allele 2 carriage in IL-1A, IL-1B, and IL-1RN in periodontitis patients who were non-smokers and in whom P. gingivalis and A. actinomycetemcomitans could not be detected (42.1 % vs. 11.3% in controls; P = 0.0068; OR 5.7, 95% CI: 1.6-19.8). Our results provide evidence that polymorphisms in genes of the IL-1 family are associated with severe adult periodontitis in the absence of other risk factors tested in this patient population.

CD14 and TLR4 Gene Polymorphisms in Adult Periodontitis

Bacterial deposits, smoking, and host genetic factors play a major role in an individual’s predisposition to periodontitis. Bacterial components are recognized by CD14 and toll-like receptor 4 (TLR4), resulting in a NF-κB-based inflammatory response. We hypothesized that functional CD14 and TLR4 polymorphisms contribute to periodontitis susceptibility. We aimed to investigate the occurrence of CD14-260C>T, TLR4 299Asp>Gly, and 399Thr>Ile gene polymorphisms in adult periodontititis. DNA was collected from 100 patients with severe periodontitis and from 99 periodontally healthy controls. The gene polymorphisms were determined by the PCR technique. The presence of the periodontal pathogens Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans, and whether the subjects smoked, was included in the analyses. The CD14-260T/T genotype was found in 34.0% of periodontitis patients and in 20.2% of controls. Logistic regression analysis adjusted for gender, age, smoking, and prevalence of P. gingivalis and A. actinomycetemcomitans showed an association between the CD14-260T/T genotype and periodontitis (P = 0.004, OR 3.0, 95% CI 1.4–6.9). We conclude that the CD14-260T/T genotype contributes to the susceptibility to severe periodontitis in Dutch Caucasians.

Clinical and microbiological effects of initial periodontal therapy in conjunction with amoxicillin and clavulanic acid in patients with adult periodontitis

The aim of the present study was to investigate the clinical and microbiological effects of initial periodontal therapy in conjunction with systemic amoxicillin plus clavulanic acid in adult periodontitis patients using a double-blind, parallel-group, and placebo-controlled protocol. 21 patients with a clinical diagnosis of generalised adult periodontitis were recruited. Clinical measurements and microbiological assessments were carried out at baseline, 3, and 12 months post-treatment. Approximately 6 weeks after initial periodontal treatment (3-6 h), patients were randomly assigned to receive coded study medication of 500 mg amoxicillin plus 125 mg clavulanic acid (Augmentin) or placebo, every 8 h for 10 days. Patients returned for follow-up visits 3, 6, 9, and 12 months after completion of the medication. The mean plaque index (PI) at baseline was 1.1 for placebo group and 0.9 for the test group. At 3 months, the PI had dropped to 0.3 in both groups, and was maintained during the rest of the study. The changes in bleeding on probing (BOP) and gingival index (GI) in the course of the study were similar in both groups. The mean whole mouth probing pocket depth (PPD) in the placebo group was 3.8 mm at baseline and 3.9 mm in the test group. A mean reduction of 1.0 mm in the placebo group and 0.9 mm in the test group was observed during the first 3 months. No further reduction in PPD was noticed during the study period in either group. There was no statistically significant difference in the PPD reduction between the 2 groups. The change in clinical attachment level (CAL) from baseline to 3 months amounted to 0.5 mm in both groups. Between 3 and 12 months, the CAL changed in neither group. In both groups, treatment resulted in a decrease in the number of spirochetes and motile rods in positive patients, but no significant differences between either group were noted in any of the dark field microscopy observations. At baseline, 1 patient in the placebo group and 2 patients in the test group were culture positive for Actinobacillus actinomycetemcomitans (Aa). After therapy, Aa was not detectable in the placebo group and 1 patient remained positive in the test group. In the placebo group, the number of patients positive for Porphyromonas gingivalis (Pg) decreased from 7 to 2 after therapy. In the test group, the 4 patients positive for Pg at baseline remained positive after therapy. In both groups, all subjects were positive for Prevotella intermedia (Pi) and Fusobacterium nucleatum (Fn) at baseline. At 12 months, all subjects had detectable subgingival Fn. 9 out of the 11 placebo and 8 of the 10 test patients remained positive for Pi. There were no differences in detection frequency of Peptostreptococcus micros (Pm) and Bacteroides forsythus (Bf) in both groups between baseline, 3, and 12 months post-treatment. The findings demonstrated that, in comparison to placebo, systemic amoxicillin plus clavulanic acid provided no additional clinical and microbiological effects in the treatment of adult periodontitis patients.

Polymorphisms in an interferon-gamma receptor-1 gene marker and susceptibility to periodontitis

Chronic marginal periodontitis is an inflammatory condition in which the supporting tissues of the teeth are destroyed. Interferon (IFN)‐γ is a cytokine that plays a pivotal role in the defense against infection, and mutations in the gene coding for the ligand binding chain (α, R1) of the IFN‐γ receptor (IFNGR1) confer susceptibility on infections caused by poorly virulent mycobacteria. Using an intronic (CA)n polymorphic microsatellite marker within the IFNGR1 gene we investigated whether genetic polymorphisms are associated with periodontitis. In 62 periodontitis patients and 56 healthy controls we found a total of 13 polymorphisms, 11 of which were found in the periodontitis patients and 9 in the controls. Although we observed a trend towards an association with disease for allele 192, there were no significant differences in allele frequency between patients and controls. We therefore cannot find any evidence to suggest that IFNGR1, as a single dominant gene, contributes to susceptibility to periodontitis. However, in combination with the environmental risk factor, smoking, the same allelic marker was significantly associated [OR = 5.56 (1.16<OR<36.31), P = 0.014, Pcorr = 0.027] with periodontitis. Our results support the multigene‐environment interaction model of disease susceptibility to periodontitis.

Host and Microbiological Factors Related to Dental Caries Development

Studies on dental caries suggest that in severe cases it may induce a systemic immune response. This occurs particularly when caries progresses into pulpal inflammation and results in abscess or fistula formation (AFF). We hypothesized that severe dental caries will affect the general health of children. The acute phase proteins alpha-1-acid glycoprotein (AGP), C-reactive protein (CRP) and the cytokine neopterin were chosen as parameters to monitor general health. Also, a polymorphism in the bacterial ligand CD14 (-260) was studied to investigate the relationship between genotype sensitivity for bacterial infections and AFF. In Suriname, children aged 6 years were recruited and enrolled into a dental care scheme, randomly assigned to 4 groups with different treatment strategies and monitored longitudinally. 348 children were included in the present study. Blood and saliva samples were taken at baseline and 1 year, and concentrations of serum AGP, CRP, neopterin, salivary Streptococcus mutans and CD14-260 C>T polymorphism were determined. There was no significant association between different treatment strategies and the serum parameters. Binary logistic regression analyses revealed a significant association between AFF as the outcome variable and the CD14 genotype and the concentrations of CRP and of neopterin as factors (p < 0.05). A significant negative association was found between the CD14-260 TT and AFF (p = 0.035, OR = 3.3) for the whole population. For children who had 4 or more carious lesions at baseline, the significance increased (p = 0.005, OR = 4.8), suggesting that the CD14-260 TT genotype was protective for AFF as a consequence of dental caries.

In vitro antimicrobial effects of two antihalitosis mouth rinses on oral pathogens and human tongue microbiota

OBJECTIVES The aim of the study was to compare the antimicrobial activity of a mouth rinse containing chlorhexidine and cetylpyridinium chloride (MR1) with a stannous fluoride-based mouth rinse (MR2) in vitro. MATERIALS AND METHODS Samples of the tongues from 10 subjects with and 10 subjects without halitosis were inoculated on blood agar plates. The agar was perforated, and the cylindrical holes were filled either with mouth rinse MR1 or with mouth rinse MR2. After incubation, inhibition zones of the whole tongue microbiota and Fusobacterium nucleatum were measured. In addition, MR1 and MR2 were applied in a short interval killing test (SIKT) on four oral pathogens Porphyromonas gingivalis, Prevotella intermedia, F. nucleatum and Aggregatibacter actinomycetemcomitans. Total viable cell counts were made after two minutes of incubation with increasing concentrations of MR1 and MR2. RESULTS MR1 showed a significantly higher in vitro antimicrobial activity against the whole tongue microbiota and F. nucleatum than MR2 in both groups of subjects. In the SIK test, MR1 showed a significantly greater killing capacity than MR2. The results show that a mouth rinse with low concentrations of chlorhexidine and 0.05% cetylpyridinium chloride appears to be more effective in inhibiting growth of the human tongue microbiota in vitro than a fluoride/stannous fluoride-containing mouth rinse. CONCLUSION This in vitro observation supports the use of chlorhexidine and cetylpyridinium chloride in the treatment of oral halitosis.

Anaerococcus nagyae sp. nov., isolated from human clinical specimens

We describe a new Anaerococcus species isolated from human clinical specimens. Analyses of 16S rRNA gene sequences of three strains showed <98% similarity with its closest relative Anaerococcus octavius. Phylogenetically the isolated strains form a cluster and can be differentiated from other species of the genus Anaerococcus based on its phenotypic characteristics and its MALDI-TOF MS profile. We propose the name Anaerococcus nagyae, with A. nagyae DSM101193 (accession number KU043522) as the type strain.

Periodontitis and Rheumatoid Arthritis

BACKGROUND Currently, in the field of rheumatology, there is much attention given towards the possible causality between periodontitis and rheumatoid arthritis (RA), specifically regarding the role of Porphyromonas gingivalis (Pg). This bacterium is unique, having a citrullinating enzyme. Antibodies against citrullinated proteins are rather specific for RA. METHODS Because causality is ultimately tested in longitudinal cohort studies which currently do not exist for periodontitis and RA, this commentary applied Bradford Hill criteria on the existing literature to assess causality as the most likely interpretation of this association. CONCLUSIONS From an epidemiologic point of view, patients with RA have a higher incidence of periodontal disease than individuals without RA. In addition, there is a dose-response pattern in the association between the severity of periodontitis and RA disease activity. There are indications that periodontitis precedes RA, but there is no evidence yet available to show that Pg plays a direct role in this temporal relationship. The role of the unique characteristic of citrullination by Pg remains unexplained. However, in animal models, citrullination by Pg plays a distinct role in the development and aggravation of experimental arthritis. Although the role of Pg in RA remains speculative, a causative role for periodontitis as a chronic inflammatory disease caused by infectious agents in RA seems biologically plausible.

Adjunctive effect of a water-cooled Nd

OBJECTIVES To test whether use of a water-cooled Nd:YAG laser adjunctive to supra- and subgingival debridement (SRP) with hand and ultrasonic instruments results in greater clinical improvement than SRP alone. Another objective was to investigate the reduction in the number of microorganisms. METHODS This study was an examiner-blind, randomized and controlled clinical trial using a split-mouth design. Nineteen subjects with moderate-to-severe generalized periodontitis were selected. Immediately following SRP in two randomly chosen contra-lateral quadrants, all pockets 4 mm were additionally treated with the Nd:YAG laser (1064 nm, 6 W, 400 mJ). Clinical assessments (Plaque index, bleeding on pocket probing, probing pocket depth) were performed pre-treatment and at 3 months post-treatment. In each quadrant, one site was sampled for microbiological evaluation at pre-treatment, immediately post-instrumentation and 3 months post-treatment. RESULTS At the 3-month visit, the clinical parameters had significantly improved for both regimens. No significant differences between treatment modalities were observed for any of the clinical parameters at any time. Immediately following instrumentation, the total colony forming units for both groups were significantly reduced as compared with pre-instrumentation. No significant differences between treatment modalities were observed. CONCLUSIONS Three months after SRP, no additional advantage was achieved with the additional use of the Nd:YAG laser. Microbiological findings reflect these clinical results.

Two-part implants inserted in a one-stage or a two-stage procedure

Objective: The aim of this study was to evaluate the feasibility of using a twopart implant system in a one-stage procedure and to monitor the microflora in the peri-implant area in relation to clinical and radiographic outcome. Material and Methods: After randomisation, 40 edentulous patients (Cawood & Howell class V–VI) received two IMZ implants in the anterior mandible inserted by either a one-stage (nΩ20) or a two-stage (nΩ20) surgical procedure for overdenture treatment. A standardised clinical and radiographic evaluation was performed after denture insertion as well as 6 and 12months thereafter. Twelve months after loading, peri-implant samples were collected and analysed for the presence of putative periodontal pathogens using culture technique. Results: No striking differences were found between the two groups with regard to the clinical parameters during the evaluation period. The mean bone loss in the first year of functioning was 0.6mm in both groups. With regard to the gingiva score, plaque score, bleeding score or bone loss between T0 and T12, no associations were found with the presence of the cultured microorganisms. An association was present between pockets 4mm and the presence of Peptostreptococcus micros in the two-stage group. Conclusions: The short-term results indicate that two-part implants inserted in a one-stage procedure may be as predictable as inserted in the common two-stage procedure. The peri-implant sulcus can and does harbour potential periodontal pathogens without significant signs of tissue breakdown.The Hawaweer, a nomadic, pastoralist group in northern Sudan, were seriously affected by the drought in the Sahel during the mid-1980s. Their experience illustrates the connection between internally displaced people, normal mobility, forced migration, dilemmas and opportunities of return and how new livelihoods can be successfully constructed based on traditional rights, strong local institutions and external resources. Some displaced Hawaweer got the chance to return to their homeland as new livelihood opportunities were established; others did not get this opportunity nor would they have returned if they had been given the chance. In both situations, the processes of displacement and return had an impact on the sense of belonging and identity.