Salivary, gingival crevicular fluid and serum levels of ghrelin and chemerin in patients with periodontitis and overweight.

BACKGROUND AND OBJECTIVE: Nutrition and body weight are modifying factors for periodontitis. The purpose of this study was to quantify two molecules (ghrelin and chemerin), released in association with food intake and obesity, in periodontally healthy and diseased individuals with respect to different body mass categories. MATERIAL AND METHODS: The two main groups (patients with chronic periodontitis and periodontally healthy/gingivitis volunteers) were subdivided into groups of subjects with normal weight [body mass index (BMI) <25] and groups of overweight/obese subjects (BMI ≥25). Subgingival bacteria were analysed and the levels of acylated and total ghrelin, chemerin and interleukin-1ß (IL-1ß) were assessed in saliva, gingival crevicular fluid and serum. RESULTS: The amount of Treponema denticola present subgingivally was significantly higher in the groups of patients with chronic periodontitis as well as in periodontally healthy/gingivitis individuals with BMI ≥25 than in periodontally healthy/gingivitis individuals with BMI <25. The amount of total ghrelin in gingival crevicular fluid differed significantly between the groups, with the lowest levels found in the group of patients with chronic periodontitis and BMI ≥25. The levels of chemerin in gingival crevicular fluid were significantly higher in each chronic periodontitis group than in periodontally healthy/gingivitis individuals with BMI <25. However, the level of IL-1ß in the gingival crevicular fluid was most differentiating between the groups, with the highest levels found in the group of patients with chronic periodontitis and BMI <25 and the lowest levels in periodontally healthy/gingivitis individuals with BMI <25. No significant differences between any groups were seen for chemerin or for acylated ghrelin in the stimulated whole saliva, or for acylated and total ghrelin in peripheral blood serum. The BMI correlated with the serum level of chemerin. CONCLUSION: Low ghrelin and high chemerin levels in the gingival crevicular fluid might be linked to periodontal disease and overweight/obesity. However, unlike IL-1ß, the levels of chemerin and ghrelin in gingival crevicular fluid are not reliable indicators of periodontal destruction.

Evaluation of a hydrophobic gel adhering to the gingiva in comparison with a standard water-soluble 1% chlorhexidine gel after scaling and root planing in patients with moderate chronic periodontitis. A randomized clinical trial.

OBJECTIVES: To evaluate the clinical, microbiological and enzymatic activity of a hydrophobic chlorhexidine-based gingiva-adhering gel containing herbal ingredients, compared with a commercially available 1% chlorhexidine water-soluble gel, during non-surgical therapy of moderate chronic periodontitis. METHODS: A total of 34 subjects participated in this 6-month blinded randomized parallel controlled trial (ISRCTN35210084). After scaling and root planing (SRP), test group received the gel, by rubbing on the gingiva, once every second day, for 14 days. The control group received the control gel twice daily. Clinical parameters considered were the approximal plaque index, simplified oral hygiene index, modified gingival index, bleeding on probing, probing depth and clinical attachment level (primary outcome), assessed at baseline, 3 and 6 months, together with the frequency of detection of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis (P.g.), Prevotella intermedia, Treponema denticola (T.d.), Tannerella forsythia (T.f.), and activity of neutrophil elastase and myeloperoxidase (secondary outcomes). RESULTS: At 3 and 6 months, all clinical parameters improved significantly, without significant intergroup differences, except OHI-S, which improved at 3 months (P < 0.05). Microbiological data resulted in no significant intergroup differences at baseline and 6 months. At 3 months, significant differences for P.g., T.f. and T.d. were noted. A significant reduction of neutrophil elastase after 3 and 6 months was observed (P < 0.005), without significant intergroup differences. For myeloperoxidase, significant reductions were noted in both groups (P < 0.005 and P < 0.05), but no significant intergroup differences. The tested product seemed to have an increased efficacy, due to longer persistence on the gingiva, with reduced application frequency. CONCLUSIONS: Both products had a relatively similar influence on the clinical, microbiological and enzymatic outcomes at 3 and 6 months after SRP.

Clinical and cytological study of the oral mucosa of smoking and non-smoking qat chewers in Yemen.

OBJECTIVE: The study was conducted to investigate the role of qat and smoking habits on the prevalence of visible and cytological abnormalities in the oral mucosa among Yemenites. METHODS: We recruited 30 non-smoking and 30 smoking Yemenites chewing qat unilaterally for at least 5 years. We inspected oral cavities for the presence of lesions and took brush biopsies from the buccal mucosa/gingiva of the chewing/non-chewing region. RESULTS: All visible oral lesions were flat and homogeneous, and cytological changes were detected frequently. Among both non-smokers and smokers, white lesions and cytological changes were detected in 77% of all cases. On the chewing area, the proportion with white lesions ranged--depending on anatomical area and smoking status--between 47 and 93% and was significantly more frequent than on the non-chewing side (range 3-47%). The proportion of regions with changes was similar in non-smokers and smokers. Kappa statistics for "interobserver" agreement between visual inspection and cytological specimens of brush biopsies was at best fair (≤0.25). CONCLUSIONS: The high prevalence of visible lesions and cytological abnormalities among qat chewers was independent of smoking status. CLINICAL RELEVANCE: The moderate level of agreement between visual inspection and exfoliative cytology demonstrates the still challenging clinical management of chronic qat chewers, though brush biopsies including adjuvant techniques like DNA cytometry may support the clinical decision-making process in future.

Antibacterial action of Chlorhexidine/thymol containing varnishes in vitro and in vivo.

OBJECTIVES: The antibacterial activity of two different formulations of a chlorhexidine/thymol varnish should be elucidated in vitro and in vivo. METHODS: The agar diffusion assay with Cervitec(®) and CervitecPlus(®) and three reference strains each of streptococci, lactobacilli, actinomyces and periodontal pathogens was performed. In a split-mouth study, 40 volunteers applied the test (CervitecPlus(®), solvent water and ethanol) and control (Cervitec(®), solvent ethyl acetate) varnish at buccal recessions of premolar teeth at baseline as well as after two, four and seven days. Supra- and subgingival plaques were collected 2 weeks before baseline and at the screening appointments. Supragingival plaque was analysed for mutans streptococci and lactobacilli and subgingival samples for Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis and Porphyromonas intermedia. Friedman/Wilcoxon tests and U-test were used for statistical analysis (P < 0.05). RESULTS: Most reference strains were susceptible with inhibition zones (mm) as follows: Cervitec(®)/CervitecPlus(®) streptococci 27 ± 1.7/21.3 ± 2.5, lactobacilli 26 ± 9.2/23.7 ± 4.9, actinomyces 36.3 ± 6.6/27.3 ± 1.5, periodontal pathogens 18.7 ± 7.6/18 ± 1.7. Both varnishes reduced significantly the counts of mutans streptococci and lactobacilli in the patients. However, no significant differences were found between test and control sides at any time. The total counts of periodontal pathogens were low. A tendency to higher counts of A. actinomycetemcomitans at the control side could be shown; the test side did not harbour significantly higher counts. CONCLUSION: Both varnishes may influence the plaque formation and reduce mutans streptococci in supragingival plaque.

The effects of stress hormones on growth of selected periodontitis related bacteria.

The focus of this study was to examine in vitro the effects of stress hormones (catecholamines: epinephrine, norepinephrine, dopamine and hydrocortisone: cortisol) on the growth of four anaerobic species of periodontitis-related bacteria (Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia and Tannerella forsythia) and one facultative anaerobic species (Eikenella corrodens). Bacterial growth was determined by two different methods: fluorescence in situ hybridization (FISH), and the viable count by culture method. To simulate stress, each single strain was grown in a special growth medium with three different concentrations of each hormone, using an anaerobic chamber at 37 °C. Growth of F. nucleatum increased in the presence of all stress hormones. Growth of P. gingivalis was not significantly influenced by any hormone. Growth of P. intermedia and E. corrodens was inhibited by almost all stress hormones tested. Both methods of analysis revealed that the highest concentrations of norepinephrine and cortisol increased the growth of T. forsythia. Different hormones have a different effect on the growth of periodontitis-related bacteria in vitro. It appears that bacterial viability is more strongly influenced than is bacterial metabolic activity. The growth of F. nucleatum particularly and partially of T. forsythia is increased by several stress hormones and may have an additional negative impact on periodontal disease.

Prospective clinical study evaluating the long-time adjunctive use of chlorhexidine after one-stage full-mouth SRP.

OBJECTIVES: Scaling and root planing are the causal procedure in the treatment of periodontitis. Many attempts have been made to improve the outcome. The aim of this study was to verify the influence of the extended use of chlorhexidine after one-stage full-mouth (FM) SRP in patients with chronic periodontitis on the clinical outcome after 3 months. METHODS: Eighty-one patients with pockets > or =5 mm were treated by FM. All patients rinsed additionally with 0.2% chlorhexidine (CHX) twice daily over 3 months. Plaque index, bleeding on probing, probing depth (PD) and clinical attachment level (CAL) were recorded at baseline and after 1 and 3 months. RESULTS: In the test group, all variables were significantly improved after 1 and 3 months. Mean reduction of PD and CAL gain was 2.25 +/- 1.08 and 1.67 +/- 1.08 after 1 and 2.99 +/- 1.11 and 2.33 +/- 1.31 after 3 months respectively. CONCLUSIONS: Over 3 months of extended use of CHX mouth rinse after SRP showed slightly but statistically significant better results.

Rapid identification of oral anaerobic bacteria cultivated from subgingival biofilm by MALDI-TOF-MS.

INTRODUCTION: To facilitate the identification of anaerobes cultivated from periodontal disease, whole cell bacterial identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was evaluated. METHODS: A total of 84 strains (nine reference strains and 75 recent clinical isolates from 33 patients with aggressive periodontitis) previously identified with phenotypic methods were used. All the references and 10 clinical isolates belonging to the same species as the reference strains were genotypically identified by sequence analysis of the 16S ribosomal RNA gene. All the strains were then analyzed using MALDI-TOF-MS. RESULTS: The reference strains of anaerobic bacteria used showed characteristic MALDI-TOF-MS spectra with peaks between m/z 2000 and up to about m/z 13,000. On visual inspection, the similarity of spectra produced by strains of a single genus could be recognized. Obvious differences between spectra produced by strains of different species were also easily noticed. The reproducibility of the method was proved by the similarity of spectra belonging to the same species. The spectra of the Prevotella intermedia strains identified with MALDI clustered together and clustered separately from the spectra of Prevotella nigrescens, proving that MALDI-TOF-MS is an accurate method that is capable of separating these two species. The quality of clustering was characterized by calculating an inconsistency coefficient (Mathworks:/Matlab Reference Manual v2007a/, Statistical toolbox). CONCLUSION: Our results suggest that MALDI-TOF-MS might become a useful method for the identification of anaerobic bacteria, especially for those that cannot be readily identified by biochemical analysis. It may become an attractive system even for the routine identification of clinical isolates.

Adjuvant treatment with interleukin-2- and interferon-alpha2a-based chemoimmunotherapy in renal cell carcinoma post tumour nephrectomy: results of a prospectively randomised trial of the German Cooperative Renal Carcinoma Chemoimmunotherapy Group (DGCIN).

We conducted a prospectively randomised clinical trial to investigate the role of adjuvant outpatient immunochemotherapy administered postoperatively in high-risk patients with renal cell carcinoma. In total, 203 renal carcinoma patients' status post radical tumour nephrectomy were stratified into three risk groups: patients with tumour extending into renal vein/vena cava or invading beyond Gerota's fascia (pT3b/c pN0 or pT4pN0), patients with locoregional lymph node infiltration (pN+), and patients after complete resection of tumour relapse or solitary metastasis (R0). Patients were randomised to undergo either (A) 8 weeks of outpatient subcutaneous interleukin-2 (sc-rIL-2), subcutaneous interferon-alpha2a (sc-rIFN-alpha2a), and intravenous 5-fluorouracil (iv-5-FU) according to the standard Atzpodien regimen (Atzpodien et al, 2004) or (B) observation. Two-, 5-, and 8-year survival rates were 81, 58, and 58% in the treatment arm, and 91, 76, and 66% in the observation arm (log rank P=0.0278), with a median follow-up of 4.3 years. Two, 5-, and 8-year relapse-free survival rates were calculated at 54, 42, and 39% in the treatment arm, and at 62, 49, and 49% in the observation arm (log rank P=0.2398). Stage-adapted subanalyses revealed no survival advantages of treatment over observation, as well. Our results established that there was no relapse-free survival benefit and the overall survival was inferior with an adjuvant 8-week-outpatient sc-rIL-2/sc-rIFN-alpha2a/iv-5-FU-based immunochemotherapy compared to observation in high-risk renal cell carcinoma patients following radical tumour nephrectomy.

Salivary analyses and caries increment over 4 years: an approach by cluster analysis.

MAIN PROBLEM: Caries as a multifactorial process is influenced by salivary defense. Cluster analyses should give additional information on the role of salivary variables in relation to caries increment. METHODS: Samples of unstimulated and stimulated whole saliva from 28 young adults (mean age 23.5+/-2.1 years) were analyzed for flow rate, pH and buffer variables, lysozyme, lactoferrin, peroxidase, thiocyanate, secretory immunoglobulin A, and total protein. The decayed, missing, and filled surfaces (DMFS) were recorded at baseline and after 4 years. Cluster analyses were executed on the basis of salivary data. RESULTS: The mean caries increment (DeltaDMFS) over 4 years was 6.7+/-4.0 (range 1-16). In two-cluster processing, three out of four volunteers with low caries increments were grouped into one cluster. Only a few variables proved to be important for cluster characteristics. CONCLUSIONS: The results suggest that over 4 years (1) the volunteers with very low caries increment (DeltaDMFS=1) are classified always together, (2) these volunteers do not form a separate cluster by themselves, (3) low caries increment was related to higher salivary flow rate and lower levels of lysozyme and lactoferrin for unstimulated saliva and (4) the partial pressure of CO(2) was of importance in stimulated saliva.

Lactoferrin and other markers from gingival crevicular fluid and saliva before and after periodontal treatment.

OBJECTIVES: The aim of the study was to verify (i) if crevicular fluid defence variables reflect the changes after surgical periodontal treatment and (ii) if they are in correspondence with changes of these variables in the unstimulated and stimulated whole saliva. MATERIAL AND METHODS: For 12 male and 13 female volunteers with chronic periodontitis lactoferrin concentration as well as the lysozyme and peroxidase activities were determined in crevicular fluid as well as in unstimulated and stimulated saliva before and 14 days after surgical periodontal treatment by a minimal invasive flap technique. RESULTS: The lactoferrin concentrations decreased significantly in the crevicular fluid eluting solution from 1.63 to 1.23 mg/l reflecting a decrease in the total amount collected, in unstimulated saliva from 10.54 to 8.96 mg/l, and in stimulated saliva from 9.00 to 7.11 mg/l after treatment. No significant change could be found for lysozyme. Peroxidase activity was significantly reduced from 269.06 to 186.15 U/l only in the crevicular fluid. CONCLUSION: The results of this study suggest that (i) the defence factor lactoferrin is suitable for monitoring of periodontal treatment results and (ii) changes of the lactoferrin concentration in crevicular fluid are related with significant changes in unstimulated and stimulated saliva.

Treatment of gingivitis with hyaluronan.

OBJECTIVES: Hyaluronic acid (hyaluronan) is a glycosaminoglycan with anti-inflammatory and antiedematous properties. It was evaluated in a gel formulation for its effect in the treatment of plaque-induced gingivitis. METHOD: In a randomised double-blind study, 50 male subjects with plaque-induced gingivitis were divided into two groups and used a verum or placebo gel twice daily additionally to oral hygiene for a 3-week treatment period. Clinical indices (API, Turesky index, PBI) and crevicular fluid variables (peroxidase, lysozyme) were determined at baseline and after 4, 7, 14 and 21 days, respectively. RESULTS: Significant improvements could be found for all clinical variables in both groups. The verum group showed significant improvement in the study area for the plaque indices beginning with day 4 (P = 0.011) and the PBI beginning with day 7 (P = 0.001) in comparison with the placebo group. The crevicular fluid variables were significantly improved in the centre of the studied inflammation area in the verum group. Here all studied sites had significant decreases in peroxidase (176.72-128.75 and 188.74-128.75 U/L) and lysozyme (1.27-0.27 and 1.30-0.33 mg/L) activities after 7, 14 and 21 days (P between 0.034 and < 0.001), whereas in the placebo group only one site showed a significant decrease for lysozyme (1.74-0.75 mg/L) after 7 and 21 days (P = 0.048 and 0.025). CONCLUSIONS: These data suggest that a hyaluronan containing gel has a beneficial effect in the treatment of plaque-induced gingivitis.

Longitudinal analysis of the association of human salivary antimicrobial agents with caries increment and cariogenic micro-organisms: a two-year cohort study.

Previous studies of the possible associations of salivary antimicrobial agents with dental caries have given controversial results, obviously mainly because almost all studies have been cross-sectional. Our aim was to find out, in a two-year longitudinal follow-up study, the associations among selected salivary non-immune and immune antimicrobial variables, cariogenic bacteria, and caries increment. The study population was comprised of 63 subjects, all of whom had their 13th birthday during the first study year. In addition to a comprehensive dental examination at baseline and after 2 yrs, paraffin-stimulated whole saliva samples were collected in a standardized way at six-month intervals. Saliva samples were analyzed for flow rate, buffer effect, lysozyme, lactoferrin, total peroxidase activity, hypothiocyanite, thiocyanate, agglutination rate, and total and specific anti-S. mutans IgA and IgG, as well as for numbers of total and mutans streptococci, lactobacilli, and total anaerobic bacteria. Cluster analysis and Spearman-Rank correlation coefficients were used to explore possible associations between and among the studied variables. During the two-year period, a statistically significant increase was observed in flow rate, thiocyanate, agglutination rate, anti-S. mutans IgA antibodies, lactobacilli, and total anaerobes, whereas lysozyme, lactoferrin, and total and anti-S. mutans IgG antibodies declined significantly. Based on various analyses, it can be concluded that, at baseline, total IgG and hypothiocyanite had an inverse relationship with subsequent two-year caries increment, anti-S. mutans IgG antibodies increased with caries development, and mutans streptococci and lactobacilli correlated positively with both baseline caries and caries increment. Total anaerobic microflora was consistently more abundant among caries-free individuals. In spite of the above associations, we conclude that none of the single antimicrobial agents as such has sufficiently strong power to have diagnostic significance in vivo with respect to future caries.

Antimicrobial factors of saliva in relation to dental caries and salivary levels of mutans streptococci.

The possible association between salivary non-immunoglobulin (lysozyme, lactoferrin, hypothiocyanite, agglutinins) or immunoglobulin (total IgA, anti-Streptococcus mutans IgA) antimicrobial factors, and the prevalence of dental caries was studied in 59 young adults. These antimicrobial factors were also analysed in relation to the salivary levels of mutans streptococci (MS). The amount of MS correlated significantly (+0.31, p less than 0.05) with the number of initial caries lesions (Di) but not with other caries indices (DMFT, DMFS, DS). The group with no Di (N = 17) had significantly (p less than 0.05) more hypothiocyanite (HOSCN/OSCN-) and anti-S. mutans IgA antibodies in whole saliva than those with initial caries lesions (N = 42). None of the antimicrobial factors alone showed any significant association with salivary MS counts. Our results suggest that HOSCN/OSCN- and anti-S. mutans IgA may be involved in the prevention of the early phases of dental caries.

[The assessment of the caries risk in young adults]. / Zur Einschätzung des Kariesrisikos bei jungen Erwachsenen.

Several factors for caries prediction have been proved in a one-year follow-up study on 73 patients. The white-spot lesions have proved to be a good criteria for caries prediction in this population. These lesions could be combined with the microbiological tests Dentocult SM and Oricult N or with the determination of the buffer capacity (Dentobuff), which did not much improve the prediction.

[The quantitative determination of resistance factors in the saliva of patients with very high caries involvement]. / Quantitativer Nachweis von Abwehrfaktoren im Speichel von Patienten mit sehr hohem Kariesbefall.

The unstimulated and stimulated whole saliva of patients with a very high caries experience were used for the determination of the flow rate, total protein, and the antibacterial proteins lysozyme, S-IgA and lactoferrin as well as the peroxidase and thiocyanate being components of the salivary peroxidase system. The concentration, the specific concentration and the secretion rate were calculated.

[The plaque wet weight in subjects with different caries involvement]. / Das Plaquefeuchtgewicht bei Probanden mit unterschiedlichem Kariesbefall.

Determinations of the wet weight of 48 h human dental plaque have proved that there is a significantly higher plaque wet weight among patients characterized by a low caries experience compared to patients with a high caries experience. The plaque-forming rate of patients with a low caries experience rises with advancing years.