Calprotectin levels in gingival crevicular fluid predict disease activity in patients treated for generalized aggressive periodontitis.

BACKGROUND AND OBJECTIVE: Clinical parameters such as probing depth and bleeding on probing are commonly used for monitoring after periodontal treatment. However, these parameters have poor prognostic utility. The biomarker calprotectin is used to monitor conditions such as inflammatory bowel disease because of its ability to predict disease activity. Levels of calprotectin in gingival crevicular fluid correlate with periodontal disease severity and treatment outcome. The validity of calprotectin as predictor for future periodontal disease activity has not yet been investigated. MATERIAL AND METHODS: Thirty-six subjects with generalized aggressive periodontitis were treated with scaling and root planing (SRP), and with adjunctive antimicrobial medications. Probing depth, clinical attachment level and bleeding on probing were assessed at baseline, and 3 and 6mo after SRP. A gingival crevicular fluid sample was collected from the initially deepest site in each patient 3mo after SRP and analysed for calprotectin levels. Activity was defined as a probing depth increase of >0.5mm between 3 and 6mo at the sample site. The ability of individual parameters to predict activity was analysed by construction of receiver operating characteristic curves. RESULTS: Nine active sites were identified. Clinical attachment level, probing depth, bleeding on probing and gingival crevicular fluid volume showed no predictive utility [area under the curve (AUC) <0.6, p>0.05]. However, calprotectin concentration (AUC=0.793, p=0.01) and the total amount/sample of calprotectin (AUC=0.776, p=0.02) significantly predicted activity. Patients with calprotectin levels above calculated cut-off values had significantly more active sites than patients with negative results. CONCLUSION: Calprotectin levels were predictors of disease activity at both site and subject levels. The calculated cut-off values provide a dichotomous basis for prospective evaluation of calprotectin as a diagnostic marker for monitoring periodontal treatment.

Periodontal disease in patients with ankylosing spondylitis.

OBJECTIVE: Ankylosing spondylitis (AS) and periodontal disease (PD) are characterised by dysregulation of the host inflammatory response, resulting in soft and hard connective tissue destruction. AS has been related to other inflammatory diseases, however, there is a paucity of data on whether AS is associated with inflammatory PD. METHODS: The association between AS and PD was examined in 48 patients with AS and 48 healthy controls, matched for age and gender. AS was diagnosed according to the modified New York criteria. Periodontal examination included probing pocket depth (PPD), clinical attachment loss (CAL), plaque index (PI) and bleeding on probing (BOP). Potential risk factors of PD such as smoking, low education, alcohol consumption, body mass index (BMI), as well as chronic diseases associated with PD and AS were assessed through questionnaires. RESULTS: In stepwise logistic regression, including AS status, age, gender, education, smoking, alcohol consumption and BMI, only AS status, age and education remained significant predictors of PD. Patients with AS had significant 6.81-fold increased odds (95% CI 1.96 to 23.67) of PD (defined as mean attachment loss >3 mm) compared to controls. The strength of the association was attenuated but remained statistically significant after further adjustment for plaque accumulation (odds ratio (OR) 5.48, 95% CI 1.37 to 22.00). CONCLUSIONS: The present study shows that patients with AS have a significantly higher risk of PD, strongly suggesting the need for close collaboration between rheumatologists, periodontists and dental hygienists when treating patients with AS.

Induced apoptosis of chondrocytes by Porphyromonas gingivalis as a possible pathway for cartilage loss in rheumatoid arthritis.

The role of bacterial infections in the pathogenesis of rheumatoid arthritis (RA) has gained increasing interest. Patients with RA often exhibit periodontal disease, which is associated with pathogens like Porphyromonas gingivalis. The present study examines the direct effects of P. gingivalis on apoptosis of human chondrocytes (a feature of inflammatory joint diseases) as one can assume an interrelation of pathogenesis of RA and P. gingivalis infections. Primary chondrocytes were infected with P. gingivalis. Early apoptotic and dead cell analysis was performed using Annexin-V, 7AAD, and propidium iodide and examined by flow cytometry and fluorescence microscopy. Caspase activation and DNA fragmentation were determined by western blot analysis and TUNEL reaction. Flow cytometry and fluorescence microscopy demonstrated an increase of Annexin-V-positive early apoptotic chondrocytes after infection. Western blot showed upregulation of activated caspase-3 expression, and TUNEL reaction revealed considerable DNA fragmentation following infection. The data show that P. gingivalis promotes early and later stages of apoptosis of primary human chondrocytes, which might contribute to the joint damage seen in the pathogenesis of RA.

Effects of Porphyromonas gingivalis on cell cycle progression and apoptosis of primary human chondrocytes.

BACKGROUND: It has been suggested that bacterial infections have a role in the pathogenesis of rheumatoid arthritis (RA). P gingivalis, a Gram-negative, anaerobic rod, is one of the major pathogens associated with periodontal disease. OBJECTIVE: To examine P gingivalis infection and its effects on cell cycle progression and apoptosis of human articular chondrocytes. METHODS: Primary human chondrocytes cultured in monolayers were challenged with P gingivalis. Infection and invasion of P gingivalis into chondrocytes was analysed by scanning electron microscopy, double immunofluorescence and by antibiotic protection and invasion assay. Cell cycle progression of infected chondrocytes was evaluated by flow cytometry. Also, cell apoptosis was visualised by terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) of DNA strand breaks and by western blot analysis. RESULTS: Data showed that P gingivalis could adhere and infect primary human chondrocytes. After chondrocyte infection, intracellular localisation of P gingivalis was noted. Flow cytometry analyses demonstrated affected cell cycle progression, with an increase of the G(1) phase and a significant decrease of the G(2) phase after infection. In addition, increased apoptosis of P gingivalis-infected chondrocytes was visualised by TUNEL assay and by upregulation of caspase-3 protein expression. CONCLUSION: These data demonstrate that P gingivalis infects primary human chondrocytes and affects cellular responses, which might contribute to the tissue damage seen in the pathogenesis of rheumatoid arthritis.

Association among rheumatoid arthritis, oral hygiene, and periodontitis.

BACKGROUND: A limited number of studies suggest a higher prevalence of periodontal disease among individuals with rheumatoid arthritis (RA); however, results have been inconsistent. Further, it is unclear to what extent poor oral hygiene among patients with RA may account for this association. METHODS: The association between RA and periodontitis was examined in 57 subjects with RA and 52 healthy controls, matched by age and gender. Oral examination included plaque index (PI), gingival index (GI), probing depth (PD), and clinical attachment loss (CAL). Potential risk factors for periodontal disease, such as smoking, education, alcohol consumption, and body mass index (BMI), as well as chronic diseases associated with RA and periodontal disease were assessed through questionnaires. RESULTS: In a stepwise logistic regression, including RA status, age, gender, education, smoking, alcohol consumption, and BMI, only RA status and age remained significant predictors of periodontal disease. Subjects with RA had a significant 8.05-fold increased odds (95% confidence interval: 2.93 to 22.09) of periodontitis compared to controls. The strength of the association was attenuated but remained statistically significant after further adjustment for PI, GI, or both. PI alone accounted for 12.4%, GI alone accounted for 11.1%, and PI and GI combined accounted for 13.4% of the association between RA and periodontitis. CONCLUSIONS: Subjects with RA have significantly increased periodontal attachment loss compared to controls. Oral hygiene may only partially account for this association.

Porphyromonas gingivalis dihydroceramides induce apoptosis in endothelial cells.

Porphyromonas gingivalis dihydroceramides are found in extracts of calculus-contaminated root surfaces, diseased gingival tissue, and atherosclerotic plaques. These ceramides have been shown to promote inflammatory secretory responses in gingival fibroblasts. Little is known about their effects on the vascular system. We tested the hypothesis that P. gingivalis lipids induce apoptosis of human endothelial cells, and investigated the effects of extracted and purified P. gingivalis lipids on human umbilical vein endothelial cells. P. gingivalis phosphoglycerol dihydroceramides induced apoptosis, but not necrosis, in endothelial cells. Early apoptotic cells showed exposure of phosphatidylserine on the cell surface, followed by the cleavage of procaspases 3, 6, and 9. The release of apoptosis-inducing factor was increased, suggesting mitochondrial involvement. Different caspase inhibitors and cAMP elevation blocked DNA fragmentation. Moreover, N-acetylcysteine significantly reduced apoptosis, suggesting a role for reactive oxygen species in this process. Analysis of these data indicates that dihydroceramides may be important virulence factors of P. gingivalis.

Obesity, inflammation, and periodontal disease.

The prevalence of obesity has increased substantially over the past decades in most industrialized countries. Obesity is a systemic disease that predisposes to a variety of co-morbidities and complications that affect overall health. Cross-sectional studies suggest that obesity is also associated with oral diseases, particularly periodontal disease, and prospective studies suggest that periodontitis may be related to cardiovascular disease. The possible causal relationship between obesity and periodontitis and potential underlying biological mechanisms remain to be established; however, the adipose tissue actively secretes a variety of cytokines and hormones that are involved in inflammatory processes, pointing toward similar pathways involved in the pathophysiology of obesity, periodontitis, and related inflammatory diseases. We provide an overview of the definition and assessment of obesity and of related chronic diseases and complications that may be important in the periodontist's office. Studies that have examined the association between obesity and periodontitis are reviewed, and adipose-tissue-derived hormones and cytokines that are involved in inflammatory processes and their relationship to periodontitis are discussed. Our aim is to raise the periodontist's awareness when treating obese individuals.

Prediction of periodontal disease from multiple self-reported items in a German practice-based sample.

BACKGROUND: Ascertainment of periodontal disease using self-reported measures would be useful for large epidemiologic studies. This study evaluates whether a combination of self-reported items with established risk factors in a predictive model can assess periodontal disease accurately. METHODS: Responses of 246 subjects to a detailed questionnaire were compared to their periodontal disease history as assessed from radiographs. Multiple regression modeling was used to construct predictive models using self-reported items and established risk factors. RESULTS: Depending on the definition of gold-standard periodontal disease, two or three self-reported items were selected for the predictive models, in addition to age, gender, and smoking. Self-reported tooth mobility was associated strongly with periodontal disease independent of other risk factors and was selected in all models. For dichotomous definitions of periodontal disease, discrimination of predictive logistic regression models was good with areas under the receiver operating characteristic curve >0.80. Assessment of periodontal disease history based on extreme quantiles of model-predicted values yielded high sensitivity and specificity. CONCLUSION: The combination of several self-reported items may be useful for ascertainment of periodontal disease in epidemiologic studies.

The double lateral bridging flap for coverage of denuded root surface: longitudinal study and clinical evaluation after 5 to 8 years.

Longitudinal results of a 5 to 8 year follow-up study of the double lateral bridging flap for coverage of gingival recessions (GR) are presented. On a total of 75 GR in 18 patients (31 surgical procedures) the majority of the teeth (58%) showed a root denudation reduction of 75% or more. Total coverage was observed on 18 teeth (24%). Examinations did not show a high correlation between the extent of recession and bone dehiscence after surgery, between the degree of root coverage and the initial width of keratinized gingiva, or between the GR before and after surgery. We conclude that neither the quantity of gingival recession nor the quality of the supporting tissues were a prerequisite for the success of this surgical technique.

Immunohistochemical distribution of extracellular matrix proteins as a diagnostic parameter in healthy and diseased gingiva.

This study showed histopathological findings of the extracellular matrix in healthy, inflamed, and hyperplastic human gingiva with immunohistochemical techniques. The distribution of collagen types V and VI, as well as of glycoprotein fibronectin, shows that they are extracellular matrix structural components which differentiate the tissue pathology. The orientation of the collagen fibers, the intensity of the fluorescent staining, the thickness of the fibrillar component, and the topographical localization of the connective tissue proteins are important parameters for tissue morphology. Therefore, bacterial deposits and the pharmacodynamic properties of drugs associated with gingival hyperplasia lead to an alteration of the matrix compared to the healthy tissues. This may be important in the tissue pathology in cases when the medical history of the patient is not known, as well as in forensic medicine.

Local metronidazole application in maintenance patients. Clinical and microbiological evaluation.

The purpose of this investigation was to evaluate the clinical and microbiological effect of local antibiotic therapy in comparison with subgingival scaling and root planing in a randomized semi-masked study. Forty-six recall patients who completed systematic periodontal therapy 6 to 24 months prior to the study were enrolled. The inclusion requirements were at least one site with probing depth > or = 5 mm in each quadrant, no scaling, and no antibiotic therapy during the last 6 months. After randomization each patient received 2 different treatments: in 2 quadrants metronidazole 25% dental gel was applied subgingivally to the pockets at day 0 and day 7; scaling and root planing was carried out in the 2 other quadrants, one at day 0 and in the remaining quadrant at day 7. Subgingival microbiological samples were taken from each patient before treatment and on days 21, 91, and 175 after the treatment. The analyses were carried out by indirect immunofluorescence assay. At all treated sites probing depth (PD), clinical attachment level (CAL), and bleeding on probing (BOP) were recorded on days 0, 21, 91, and 175. Both treatments resulted in PD reduction and CAL gain. PD reduction was statistically significant (P < 0.01) for both treatment modalities after 6 months. The CAL gain was not significant for either treatment. There was no statistical significance between scaling and antibiotic therapy. Treponema denticola, Porphyromonas gingivalis, and Prevotella intermedia were significantly reduced after therapy; however, there were no statistically significant differences between treatments. If Actinobacillus actinomycetemcomitans was present before therapy, it was also present after treatment in both groups. The conclusion is that, in recall patients, local application of metronidazole and scaling and root planing showed similar clinical and microbiological effects without statistically significant differences.

Local versus systemic adjunctive antibiotic therapy in 28 patients with generalized aggressive periodontitis.

BACKGROUND: Although the use of systemic antibiotics has been studied in patients with generalized aggressive periodontitis (formerly rapidly progressive periodontitis), the use of adjunctive tetracycline fibers in these patients has not been reported. The purpose of the present study was to compare the clinical response of local versus systemic antibiotic treatment as adjuncts to scaling and root planing in patients with GAgP. METHODS: After initial therapy and full-mouth scaling and root planing (SRP), 30 patients were randomly assigned to 1 of 2 antibiotic treatment groups. Probing depth (PD), clinical attachment level (CAL), and bleeding on probing (BOP) were recorded with an automated probe prior to SRP at baseline (BL) and 15, 30, 41, and 54 weeks later. Three months after SRP, the patients were treated with amoxicillin/clavulanic acid (500 mg tid; SRP + AUG group) or with local tetracycline fiber in pockets with PD > or =5 mm (SRP + TCF group). RESULTS: In both treatment groups, PD decreased significantly from BL to week 54 (6.2+/-1.5 mm to 4.7+/-1.4 mm for SRP + TCF and 6.5+/-1.4 mm to 4.2+/-0.6 mm for SRP + AUG). However, there was no statistically significant difference between the 2 groups in pocket reduction. Similarly, in both treatment groups, there were small but significant gains in CAL from BL to week 54 (12.0+/-1.8 mm to 11.3+/-1.8 mm for SRP + TCF and 12.3+/-1.5 mm to 11.2+/-1.2 mm for SRP + AUG). The difference in CAL gain between the 2 groups was not statistically significant. At the final examination, both groups showed significant PD reduction and CAL gain (P <0.001) compared to BL. The frequency and percentage of bleeding sites decreased significantly in both groups. At week 54, this decrease was significantly greater in the SRP + AUG group (31.67% for SRP + TCF versus 3.85% for SRP + AUG). CONCLUSIONS: These results indicate that the local delivery of tetracycline by a fiber or the systemic administration of amoxicillin/clavulanic acid given 3 months after scaling and root planing produced similar clinical outcomes over the 9-month observation period.

Observations on a new collagen barrier membrane in 16 consecutively treated patients. Clinical and histological findings.

BACKGROUND: Space-maintaining capacity, cell disclusive potential, and stability over time are crucial factors to achieving sufficient bone augmentation with membrane barriers. The case series presented here assessed a new collagen barrier used in bone augmentation. Clinically, the healing pattern, especially in cases of secondary healing, was studied. METHODS: Soft tissue healing was documented by photographs, and the size of the dehiscences calculated by image analysis. The measurements were performed on digitized photographs. During reentry, barrier remnants were dissected and histologically evaluated. RESULTS: The mean value for dehiscences was 35.5 mm2; all dehiscences healed within 4 weeks after the exposure became evident. The difference was statistically significant between the week 2 and week 6 visits (P = 0.008) for each previously exposed site. The histologic observation of barrier remnants revealed direct apposition of fibrous and bone tissues on the membrane surface. CONCLUSION: In cases of membrane exposure, gingival dehiscences always disappeared in the following weeks without affecting the healing process. Histologic results showed barrier stability over a 6-month period, promoting bone regeneration.

Changes in the differentiation pattern of oral mucosal epithelium following heterotopic connective tissue transplantation in man.

Anterior palatal connective tissue was transplanted into wound beds created within the alveolar mucosa of the lower anterior vestibulum in seven adults females. Six months postoperatively, the epithelial lining of the inverted transplants was biopsied and compared histologically, histometrically, and stereologically to the neighboring mucosal epithelium. The data suggest that (1) connective tissue of the lamina propria is primarily responsible for the structural specificity of the overlying epithelium, (2) inductive stimuli are not confined to superficial layers but also originate from deep portions of the lamina propria, (3) these stimuli influence both the epithelial differentiation and the type of epithelium-connective tissue interface, and (4) the epithelial response to the inductive stimuli in man may depend on the individual type of epithelial differentiation pattern originally prevailing at the recipient site. The transplant-lining epithelium assumed features resembling that of the hard palate in subjects with an alveolar mucosal epithelium with cheek-type characteristics. In subjects with an alveolar epithelium displaying a mixed differentiation pattern, the transplant-lining epithelium continued to generate a mixed pattern or else shifted to more palate-like characteristics.

Suppression of interleukin-10 release from human periodontal ligament cells by interleukin-1beta in vitro.

Periodontitis is characterized by an inflammatory process induced by periodontopathogenic bacteria in the subgingival plaque. Periodontal inflammation can be enhanced by both an increase of inflammatory stimulators, e.g. interleukin (IL)-6, and a decrease of inflammatory inhibitors, e.g. IL-10. The amount of IL-1beta is known to be increased in gingival tissues and in the gingival crevicular fluid from inflamed sites compared to healthy sites. This in vitro study sought to clarity whether IL-1beta (1 ng/ml) has a regulatory effect on the release of these two cytokines from human periodontal ligament (PDL) cells. PDL cells derived from healthy premolars were grown in the presence and absence (control) of IL-1beta. The concentration of IL-6 and IL-10 in the supernatants was assessed by enzyme-linked immunosorbent assay after 48 h of culture. PDL cells incubated with IL-1beta released significantly (p < 0.05) higher amounts of IL-6 and significantly (p < 0.01) smaller amounts of IL-10 compared to control. These results give further support to the observation that IL-1beta can increase the IL-6 secretion from PDL cells. Moreover, they provide original evidence that PDL cells secrete IL-10, which can be suppressed by IL-1beta. It is concluded that PDL cells can function as accessory immunoinflammatory cells amplifying the inflammatory process in periodontitis and, thereby, contributing to periodontal breakdown.

Guided bone regeneration next to osseointegrated implants in humans.

The purpose of this investigation was to evaluate the technique of guided tissue regeneration on a clinical and histologic level for generating new bone adjacent to osseointegrated implants. Cylindrical, press-fitted, titanium plasma flame-coated implants were modified by a disk screwed onto the implant. After implant insertion, the disk-implant assembly was covered by an expanded polytetrafluorethylene membrane. In one group, the space underneath the membrane was filled with hydroxylapatite granules; in the control group, no membrane was used. Clinical and histologic results demonstrate that the principle of guided tissue regeneration is applicable to osteogenesis in humans.

A clinical study comparing two high-fluoride dentifrices for the treatment of dentinal hypersensitivity.

Amine fluoride, the active ingredient of a currently marketed dentifrice in Germany and other European countries, and sodium fluoride were compared to a placebo dentifrice for their effectiveness in alleviating dentinal hypersensitivity. This was a randomized, double-blind, two-center, parallel clinical study covering 8 weeks of product use by 115 subjects. The hypersensitivity of the affected teeth was assessed by tactile stimulation, cold air stimulation, and overall subjective patient response. The three treatment groups exhibited comparable baseline sensitivity. These three methods of clinical assessment demonstrated that the desensitizing ability of a relatively higher fluoride dentifrice (1,400 ppm), delivered either as amine fluoride or sodium fluoride, did not differ significantly from that of the placebo dentifrice.

[The distribution of different collagen types and laminin in the gingiva of the common marmoset (Callithrix jacchus)]. / Die Verteilung verschiedener Kollagentypen und des Laminins in der Gingiva von Marmoset-Affen (Callithrix Jacchus).

The distribution of the collagen types I, III, IV, V, VI and of the glycoprotein laminin have been examined in the healthy gingiva of the marmoset with the indirect immunofluorescence method. It has been shown that collagen type I formed thick fibrils in a wave-like pattern, while collagen type III formed thinner fibrils, especially under the epithelial basal membrane. Collagen type IV and laminin showed a strong fluorescence in the basal membrane (epithelium, vessels, nerves). Collagen type V has been demonstrated as a fine fibrillar pattern in the lamina propria and occasionally close to the basal membrane. Collagen type VI has been represented as a microfibrillar network in the connective tissue matrix with a brilliant pericellular fluorescence around vessels and nerves. The study showed the structural model of the connective tissue matrix of the marmoset gingiva and allows a later comparison between inflamed and healthy gingiva.

Clinical and inflammatory effects of galvano-ceramic and metal-ceramic crowns on periodontal tissues.

Prosthetic crowns made by galvano-forming are considered to be highly biocompatible and aesthetic restorations. Therefore, they represent an alternative crown-system to conventional metal-ceramic crowns in replacing lost tooth structure. However, there are few data available on clinical and biochemical effects of galvano-ceramic crowns on periodontal tissues. The purpose of this controlled study was to test the impact of galvano-ceramic crowns and metal-ceramic crowns on clinical and inflammatory responses of periodontal tissues. A prospective, blinded randomized clinical trial was conducted. Galvano-ceramic crowns and metal-ceramic crowns were placed in 52 periodontally healthy patients in split-mouth design. Clinical parameters (gingival index, plaque index, probing depths and recessions) were recorded from six sites per tooth. Initial tissue alteration was accessed analysing the gingival crevicular fluid flow rate and IgG concentration in gingival crevicular fluid. After 24 months, 34 patients could be re-evaluated. All crowns were in adequate function and obvious clinical inflammation was rarely observed. After 24 months of follow-up, gingival tissues adjacent to galvano-ceramic crowns showed significantly less signs of clinical and inflammatory responses according to plaque index (P = 0.004), gingival index (P < 0.001), gingival crevicular fluid flow rate (P = 0.012) and IgG (P = 0.002). Data were also analyzed for buccal and oral sites separately. Gingival tissues adjacent to metal-ceramic crowns showed significantly increased clinical and inflammatory values for plaque index (P = 0.005), gingival index (P = 0.008), gingival crevicular fluid flow rate (P = 0.006), IgG (P = 0.007) at oral sites compared to galvano-ceramic crowns. Our data suggest a stabilizing effect of galvano-ceramic crowns on periodontal tissues over time.