benzoyl-DL-arginine-naphthylamide Research Articles

Effect of chlorhexidine chip in the treatment of chronic periodontitis

Aims:The evaluation of clinical and specific microbiological changes associated with chlorhexidine chip in the chronic periodontitis patients.Materials and Methods:A total of 30 chronic periodontitis patients (aged 20-65 years) having pocket depth of ≥5 mm in molar teeth were selected and randomly divided into following treatment groups: Group 1 - Scaling and root planning (SRP), Group 2 - SRP along with chlorhexidine chip and Group 3 - Only chlorhexidine chip. The clinical and microbial parameters were recorded at baseline and 1 and 3 months post-treatment as above. Benzoyl-DL-arginine-naphthylamide (BANA) chair side test was used for estimation of specific microbiota.Results:Gingival index, probing pocket depth and clinical attachment level scores in selected teeth within the groups at different time intervals was significantly (P < 0.001) different. Although, the comparison between groups for specific microbiota in selected sites at different intervals was not statistically significant at baseline and 1 month, it reached statistical significance at 3rd month post-treatment. Although significant reductions in percentage of BANA positive sites were observed in all three groups, the Group 2 had significantly greater percentage of BANA negative sites.Conclusion:Local drug delivery using chlorhexidine chip enhances the benefit of SRP in the treatment of chronic periodontitis.

"Investigation of the Periodontal and Microbiological Status of Patients Undergoing Fixed Orthodontic Therapy"

The aim of the study was to investigate the periodontal and microbiological status of patient undergoing fixed orthodontic treatment. Methods: In this study, plaque samples were collected from 12 patients at baseline, four monthly visits during orthodontic treatment and at 30 days after removal of the appliance. A benzoyl-DL-arginine-naphthylamide (BANA) test was performed to identify the periodontal pathogens. Dark field microscopy was used to recognize the morphotypes. The Plaque Index and probing depths were assessed at each test interval to determine the hygiene and periodontal status of the patients. Data were analyzed using analysis of variance and Tukey's test. Results: Significant increase in plaque score, probing depths, and BANA scores were found at each interval after placement of orthodontic appliances. The levels, however returned to baseline after removal of the appliances. Dark field microscopy confirmed increases in small spirochetes (8.5%), large spirochetes (2%), non-motile rods (8.5%), fusiforms (5.5%), and filaments (1%) with orthodontic treatment. Conclusions: Patients undergoing orthodontic therapy have an increase in plaque accumulation, probing depth, and microbial activity that may be associated with periodontal destruction. Thirty days after removal of the orthodontic appliance, the plaque score, probing depth, and BANA test score returned to almost baseline level.

Aterosclerosis en sujetos con periodontitis

Chronic inflammation and infections are involved in the development and progression of atherosclerotic vascular disease.To evaluate the association between periodontitis and early atherosclerosis.Fifty-three subjects who received periodontal treatment and regular maintenance for at least 10 years, and 55 subjects with periodontitis but without a history of periodontal treatment were studied. Carotid artery intima-media wall thickness (CIMT) was measured with high-resolution B-mode ultrasonography. A blood sample was obtained to measure high sensitivity C-reactive protein, fibrinogen, lipoprotein cholesterol, leukocyte count and erythrocyte sedimentation rate. Covariates included age, gender, smoking, level of education, body mass index and physical activity. The benzoyl-DL-arginine-naphthylamide (BANA) test was used to determine the number of periodontal sites with periodontal pathogens.CIMT value was significantly higher in subjects with periodontitis than those without it (0.775 ± 0.268 and 0.683 ± 0.131 mm respectively, p = 0.027). C-reactive protein, leukocyte count and percentage of sites with periodontal pathogens were also significantly higher in subjects with periodontitis. Regression analysis identified age, periodontitis, and smoking as independent predictors of CIMT.These results suggest that untreated periodontitis is associated with early atherosclerotic carotid lesions and higher levels of inflammatory markers.

Association between atherosclerosis and periodontitis

Chronic inflammation and infections are involved in the development and progression of atherosclerotic vascular disease. To evaluate the association between periodontitis and early atherosclerosis. Fifty-three subjects who received periodontal treatment and regular maintenance for at least 10 years, and 55 subjects with periodontitis but without a history of periodontal treatment were studied. Carotid artery intima-media wall thickness (CIMT) was measured with high-resolution B-mode ultrasonography. A blood sample was obtained to measure high sensitivity C-reactive protein, fibrinogen, lipoprotein cholesterol, leukocyte count and erythrocyte sedimentation rate. Covariates included age, gender, smoking, level of education, body mass index and physical activity. The benzoyl-DL-arginine-naphthylamide (BANA) test was used to determine the number of periodontal sites with periodontal pathogens. CIMT value was significantly higher in subjects with periodontitis than those without it (0.775 ± 0.268 and 0.683 ± 0.131 mm respectively, p = 0.027). C-reactive protein, leukocyte count and percentage of sites with periodontal pathogens were also significantly higher in subjects with periodontitis. Regression analysis identified age, periodontitis, and smoking as independent predictors of CIMT. These results suggest that untreated periodontitis is associated with early atherosclerotic carotid lesions and higher levels of inflammatory markers.

Periodontal Disease Activity Measured by the Benzoyl‐DL‐Arginine‐Naphthylamide Test Is Associated With Preterm Births

Infection is a risk factor for preterm birth. This study was conducted in the field and addressed the link between periodontal pathogens measured with the benzoyl-DL-arginine-naphthylamide (BANA) test and preterm birth. This prospective study was performed in Changhua, Taiwan. Periodontal examinations included the plaque index, papillary bleeding scores, and measurement of the BANA enzyme in plaque samples at the second and third trimesters. Independent variables included maternal demographic characteristics, previous pregnancy histories, risk factors, plaque and gingivitis scores, and current pregnancy outcomes. There were 19 (7%) preterm deliveries among the 268 subjects. A history of a previous preterm birth and low birth weight, frequency of prenatal visits, preterm uterine contractions, antepartum hemorrhages, placenta previae, and preterm premature rupture of membranes were significantly related to preterm birth (P = 0.035, 0.027, <0.001, 0.025, 0.006, 0.014, and <0.001, respectively). Maternal weight gain was higher with a normal term delivery (P = 0.003). Multivariable logistic regression analyses showed that the number of BANA-infected sites in the third trimester (odds ratio [OR]: 5.89; 95% confidence interval [CI]: 1.5 to 31.6), maternal weight gain (OR: 0.78; 95% CI: 0.65 to 0.91), antepartum hemorrhages (OR: 10.0; 95% CI: 2.2 to 46.9), and preterm premature rupture of membranes (OR: 12.6; 95% CI: 3.97 to 42.71) had significant influences on preterm-birth outcomes. BANA-positive plaque in the third trimester was associated with preterm births after controlling for other risk factors. The BANA test can be used to screen pregnant women at chairside and/or bedside to apply suitable intervention tactics.

The Diagnosis of Periodontal Disease in Private Practice

Periodontal disease is an inflammatory reaction to the bacteria in dental plaque. The present study compared the prevalence of periodontal disease in patients using as a diagnostic either probing depth measurements, an inflammatory marker such as numbers of white blood cells in plaque samples, or microbiological markers such as the microscopic count and the benzoyl-DL-arginine naphthylamide (BANA) test. Teeth with the most inflammation and/or deepest pockets in each quadrant were probed and subgingival plaque was sampled from 1,043 consecutive new patients enrolled in a private practice. Multivariate "diagnostic" models were developed based upon the probing depth (general linear models), percentage of white blood cell-positive and percentage of BANA-positive plaques (logistic regression models) to determine the prevalence of patients with periodontal disease. Plaque samples were removed from 3,694 sites. Fifty-two percent of sampled pockets were >4 mm; 49% of sites were inflamed, using the presence of white blood cells, and 28% were infected using the BANA test. Diagnostic models were highly significant at P<0.0001. The white blood cell model was the most parsimonious as demonstrated by the lowest Akaike information criteria statistic and had the highest receiver operator characteristic (ROC) curve relative to the probing depth and BANA models. Periodontal disease can be diagnosed chairside by the presence of white blood cells in plaque samples, a finding that reflects the inflammatory nature of the disease process. This approach would reduce the misclassification of subjects as having periodontal disease (130 patients in the present study who had pockets) but minimal evidence of an inflammatory response.

Systemic Inflammatory Markers, Periodontal Diseases, and Periodontal Infections in an Elderly Population

To study the levels of systemic markers for inflammation with parameters of periodontal diseases in older people. A cross-sectional study was conducted in a cohort that is being followed prospectively on the effects of aging and body composition on morbidity. University of Pittsburgh, Pittsburgh, and University of Tennessee, Memphis. One thousand one hundred thirty-one participants (mean age+/-standard deviation 72.7+/-2.8); 66% white and 50% male. Periodontal examination, including probing depth and attachment loss, was performed. Periodontal disease extent was divided into 0% of sites with probing depth of 6 mm or more, 1% to 10% of sites with probing depth of 6 mm or more and more than 10% of sites with probing depth of 6 mm or more. Subgingival plaque samples were collected from four molar teeth, and the levels of periodontal pathogens were determined using the benzoyl-DL-arginine-naphthylamide (BANA) test. Plasma interleukin-6 (IL-6), C-reactive protein (CRP), plasminogen activator inhibitor type-1 (PAI-1), and tumor necrosis factor alpha (TNF-alpha) levels were measured in all participants. Assessments of risk factors associated with elevated levels of markers of systemic inflammation were also determined. Multiple regression analysis was employed to analyze the data. IL-6 levels were significantly higher in participants with more-extensive periodontal disease than in other participants. Periodontal disease extent was significantly associated with higher TNF-alpha plasma levels, controlling for established risk factors for elevated TNF-alpha levels. Participants with BANA-positive species had significantly higher CRP plasma levels when controlling for risk factors for elevated CRP levels. Periodontal disease and infection may be modifiable risk indicators for elevated levels of systemic inflammatory markers in older people.

Effect of periodic preventive care on the progression of periodontal disease in young adults with Down's syndrome

To examine the effect of periodic preventive care on the progression of periodontal disease in 24 young adults with Down's syndrome (DS) (mean age+/-SD: 20.8+/-5.6 years) Subjects were divided into two groups: 13 subjects who had frequently visited our clinic (mean interval between visits: 3.7+/-1.3 months; managed group) and 11 subjects who had not visited our clinic for more than 1 year (mean duration of no visits: 27.5+/-10.1 months; interrupted group). The progression of periodontal disease in the subjects was evaluated clinically, microbiologically and roentgenologically. Clinical parameters (probing depth, frequency of the presence of a pathological periodontal pocket, alveolar bone loss, frequency of the incidence of pathological bone loss, and modified total PMA (M-PMA)) and benzoyl-DL-arginine-naphthylamide (BANA) scores in the interrupted group were significantly higher than those in the managed group. Subject age showed significant positive correlations with probing depth and alveolar bone loss. BANA scores showed significant positive correlations with probing depth, alveolar bone loss and M-PMA in the interrupted group. The results suggest that periodic preventive care is effective for suppressing the progression of periodontal disease in young adults with DS.

Relationship of sulcular sulfide level to severity of periodontal disease and BANA test.

Volatile sulfur compounds (VSC), such as hydrogen sulfide and methyl mercaptan, are toxic metabolites produced by periodontal pathogens. Their relationship to periodontal disease severity is not yet fully understood. Hence, the aims of this study were to: 1) examine the relationship between sulcular sulfide (pS) levels and severity of periodontal disease and 2) examine the link between pS level and the BANA (benzoyl-DL-arginine-naphthylamide) test. Seventy systemically healthy subjects with a mean age of 53.0 +/- 13.8 years participated. Three sites were selected from each subject based upon radiographic bone loss (RBL): RBL < 2 mm, healthy; RBL > or = 2 to < 4, low to moderate; RBL > or = 4 mm, severe. Periodontal parameters, probing depth (PD), clinical attachment level (CAL), and bleeding on probing (BOP), were recorded. The pS level was measured using a portable sulfide monitor in a digital score ranging from 0.0 (< 10(-7) M of S) to 5.0 (> or = 10(-2) M of S) in increments of 0.5. The presence of specific bacteria in subgingival plaque was detected using BANA test. The mean pS level was 0.10 +/- 0.23, 0.36 +/- 0.48, and 1.10 +/- 0.87 for healthy, low to moderate, and severe disease sites, respectively, and was statistically different (P<0.001). The pS level was positively correlated with the BANA test, and was higher in untreated subjects than maintenance subjects (P<0.01). The pS level may be a potential indicator for detecting severity of periodontal disease and identifying bacteria that are capable of hydrolyzing BANA.

Relationship between sulcular sulfide level and oral malodor in subjects with periodontal disease.

The relationship between oral malodor and sulfide levels in periodontal pockets (pS) has not yet been determined. The aims of this study were: 1) to identify the correlation among oral malodor, pS levels, and the BANA (benzoyl-DL-arginine-naphthylamide) test and 2) to recognize the interaction between oral malodor, demographic factors, tongue coating, and periodontal condition. Eighty-one periodontal patients participated in this study. A portable sulfide monitor and organoleptic method were used to evaluate oral malodor. Demographic data included age, gender, race, and smoking habits. The volume of tongue coating and periodontal condition for all teeth were assessed. The pS levels of 3 different radiographic bone loss (RBL) sites: RBL < 2 mm, healthy; RBL > or = 2 to < 4 mm; low to moderate; RBL > or = 4 mm, severe, were measured using an industrial sulcular sulfide-monitoring device. Subgingival plaque samples from the above 3 sites and tongue scraping were examined by the BANA test. The volume of tongue coating (P<0.001), extent of periodontal disease (P<0.05), pS levels of the sites with low to moderate bone loss (P<0.05), and BANA score of tongue scrapings (P<0.05) were significantly associated with oral malodor. Stepwise multiple regression analysis examined the degree of association between oral malodor and potential explanatory variables. The volume of tongue coating and percent of sites BOP (bleeding on probing) were significantly associated with oral malodor. Females and smoking habit were negatively correlated with organoleptic measurements. The pS level of the representative sites with low to moderate bone loss demonstrated a modest association with oral malodor. Oral malodor in periodontal patients was primarily associated with tongue coating and gingival inflammation.

The relationship between microbial factors and gingival crevicular fluid glycosaminoglycans in human adult periodontitis

Counts of cultivable Porphyromonas gingivalis, assays of microbial proteases and the concentration in gingival crevicular fluid of proteoglycan metabolites were investigated at periodontitis and gingivitis sites in 16 patients with chronic adult periodontitis before and after treatment. Two periodontitis sites and two gingivitis sites were selected from each patient on the basis of a clinical examination. Gingival crevicular fluid from each site was analysed for the concentrations of the glycosaminoglycans chondroitin-4-sulphate and hyaluronan and subgingival plaque samples were analysed for cultivable P. gingivalis and microbial trypsin-like proteases assayed by benzoyl- dl-arginine-naphthylamide (BANA) hydrolysis. Significantly higher concentrations ( p = 0.007) of chondroitin-4-sulphate were found at periodontitis than gingivitis sites but there was no significant difference in hyaluronan ( p = 0.36) between these sites. Although the majority of periodontal sites were P. gingivalis-negative (23/32), there were significantly higher concentrations of chondroitin-4-sulphate ( p = 0.05) and hyaluronan ( p = 0.04) at the P. gingivalis-positive, compared to -negative, periodontitis sites. At BANA-positive periodontitis sites there were also higher concentrations of chondroitin-4-sulphate ( p = 0.0015) and hyaluronan ( p = 0.0001) than at BANA-positive gingivitis sites. There was a significant decrease in concentrations of chondroitin-4-sulphate and hyaluronan at periodontitis sites after treatment. This study lends support to the hypothesis that P. gingivalis may be actively involved in the destruction of connective tissue components at culture-positive sites but shows that elevated concentrations of connective tissue breakdown products may occur in gingival crevicular fluid from periodontal sites where this organism is absent.

Presence of Treponema denticola and Porphyromonas gingivalis in children correlated with periodontal disease of their parents.

Considerable evidence exists suggesting that periodontal disease is due to the overgrowth of a finite number of specific bacteria such as Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Treponema denticola, Bacteroides forsythus, and Prevotella intermedia, among others. Three of these organisms-P. gingivalis, T. denticola, and B. forsythus-can be easily detected in plaque samples by the hydrolysis of the synthetic trypsin substrate benzoyl-DL-arginine-naphthylamide (BANA). The aim of the present study was to determine if a relationship could be found between the presence of either these organisms of periodontitis in the parent and the presence of BANA-positive species in the child. Thirty-four mothers or fathers and 34 children were examined for plaque scores, papillary bleeding scores, and the presence of P. gingivalis and T. denticola in four subgingival or marginal gingival plaque samples as assayed by the BANA test or specific polyclonal antibodies using an ELISA. Children whose parents were colonized by BANA-positive bacteria were 9.8 times more likely to be colonized by these BANA-positive species. Children whose parents had clinical evidence of periodontitis were 12 times more likely to be colonized by these BANA-positive species. These data are compatible with the hypothesis that children may acquire the BANA-positive species from their parents, especially if the parent has periodontitis.

The Utility of the BANA Test for Monitoring Anaerobic Infections due to Spirochetes (Treponema denticola) in Periodontal Disease

Treponema denticola, Porphyromonas gingivalis, and Bacteroides forsythus each possesses an enzyme(s) that hydrolyzes the synthetic substrate benzoyl-DL-arginine-naphthylamide (BANA). The presence of these organisms in a subgingival plaque sample can be determined by the ability of the plaque to hydrolyze BANA. In the present study, we describe the usefulness of the BANA test at various stages of a clinical trial of the efficacy of metronidazole in the treatment of periodontal disease. A BANA-positive test was significantly associated with high levels and proportions of spirochetes in the plaque, so that it provided information comparable with that which could be obtained by a microscopic examination of the plaque. Patients with such anaerobic spirochetal infections were randomly assigned to a group receiving either metronidazole or placebo (250 mg, three times a day) for one week and whose teeth were scaled and root-planed. The advantages of the decision that metronidazole be used were apparent from the comparison with the results obtained in the patients who received only the scaling and root planing. The initially BANA-positive teeth in the patients treated with metronidazole, scaling, and root planing gained attachment and exhibited a significant reduction in the need for periodontal surgery, when compared with the BANA-positive teeth in the patients who received only placebo, scaling, and root planing. After the conclusion of this therapy, those teeth with persistent BANA-positive plaques had significantly higher proportions and levels of spirochetes than did the teeth with BANA-negative plaques.(ABSTRACT TRUNCATED AT 250 WORDS)

Development of a diagnostic test for anaerobic periodontal infections based on plaque hydrolysis of benzoyl-DL-arginine-naphthylamide

Treponema denticola, Porphyromonas (Bacteroides) gingivalis, and Bacteroides forsythus are among the anaerobic species frequently associated with adult forms of periodontal disease. These organisms hydrolyze the synthetic peptide benzoyl-DL-arginine-naphthylamide (BANA), and such enzyme activity can be detected in the plaque and related to clinical disease and the presence of spirochetes. In this investigation, the liquid BANA assay was compared with a commercially developed BANA assay which employed a paper format and which could be read after a 15-min incubation. In the paper format, strips of a Whatman filter paper were impregnated with BANA and strips of nitrocellulose paper were impregnated with fast black K salt. Both strips were applied lengthwise across a paper card (3 by 5 in. [7.6 by 12.7 cm]). The BANA strip at the bottom was inoculated with the test sample (pure culture, plaque), folded back so that it contacted the fast black strip, and then incubated for 15 min at 55 degrees C. T. denticola, P. gingivalis, and B. forsythus always gave a positive reaction, whereas 51 other plaque species were always negative. Six Bacteroides and Capnocytophaga species on occasion had weak reactions. The proportional agreement between BANA positiveness and clinical disease was similar for both the liquid and the paper assays. The sensitivity, specificity, and accuracy relative to the clinical standard of the liquid assay were 74, 76, and 77%, respectively, while those of the paper assay were 81, 78, and 80%, respectively. The paper assay was significantly associated with the presence of either T. denticola or P. gingivalis or both in the plaque samples, with a sensitivity of 85%, a specificity of 53%, and an accuracy of 79%. These findings indicate that a rapid paper assay for BANA hydrolysis gives data comparable to those obtained with the liquid BANA assay.