Association of F. alocis and D. pneumosintes with Periodontitis Disease Severity and Red Complex Bacteria.

Oral biofilms are considered the principal etiological agent in the development of periodontitis. Novel species that may contribute to periodontitis and dysbiosis have been identified recently. The study aims to evaluate the presence of F. alocis and D. pneumosintes in healthy and diseased patients and their association with clinical parameters and with red complex bacteria. The study included 60 subjects, with 30 patients each in the healthy and periodontitis groups. The clinical parameters were noted, and samples were subjected to DNA extraction followed by a polymerase chain reaction. Statistical analysis was performed using the Graph Pad Prism software. Results: F. alocis and D. pneumosintes were detected at a significantly higher percentage in the periodontitis group compared to the healthy group (p < 0.05). D. pneumosintes was significantly associated with T. forsythia in the periodontitis group (p < 0.05). Both of these organisms were present in sites with higher clinical attachment loss (p < 0.05). This study demonstrated that both F. alocis and D. pneumosintes were detected at a significantly higher percentage in periodontitis subjects and were detected more frequently in sites with a greater clinical attachment loss. It was also evident that both F. alocis and D. pneumosintes can be present independently of other putative periodontal pathogens.

Isolation and immunohistochemical characterization of periodontal ligament stem cells: A preliminary study.

CONTEXT: It is a known fact that periodontal tissue regeneration can be achieved by the use of periodontal ligament stem cells (PDLSCs). Current mainstay of periodontal treatment is focusing on stem cell tissue engineering as an effective therapy, making it important to isolate PDLSCs from periodontal tissues. AIMS: The present research endeavor was undertaken to elucidate a technique for isolating PDLSCs for in vivo reconstructing the natural PDL tissue. SETTINGS AND DESIGN: The study design involves In vitro prospective study. MATERIALS AND METHODS: Premolar teeth were extracted from 12 patients who were under orthodontic treatment. PDL cells were scraped from their roots. Using 10 ml of Dulbecco's modified Eagle's medium with pH 7.2, the specimens of the periodontal tissue were transferred to laboratory where cell culture was done. Isolated stem cells were grown on 24-well microtiter plates-containing cover slips. They were incubated overnight at approximately 37°C in 95% air and 5% humidification. Anti-CD 45, CD73, CD90, CD105, and CD146 antibodies were used. After staining, cells were observed under phase-contrast microscopy and in inverted microscope. RESULTS: The cells showed a marked growth and 90% confluence at day 6. Cells presented thin and long fibroblastic spindle morphology. Isolated PDLSCs showed colony-forming ability at the 14th day after seeding. Immunohistochemical staining of PDLSCs showed positive uptake for CD146, CD90, CD73, CD105, and negative uptake for CD45. CONCLUSIONS: The human PDLSCs can be clearly isolated and characterized by using CD90, CD73, CD146, and CD105 markers of stem cells.

Interleukin-21 and its association with chronic periodontitis.

CONTEXT: Interleukin-21 (IL-21) is a pleiotropic cytokine, well documented to contribute to the development of Th17 cells which have been shown to play an important role in the pathogenesis of periodontitis. Periodontal disease is a chronic infection of tooth-supporting tissue. AIM: This study evaluates the saliva and serum levels of IL-21 in patients with chronic periodontitis and periodontally healthy individuals. SETTINGS AND DESIGN: The present study was carried out in the Department of Microbiology in association with Department of Oral Medicine and Radiology, Maratha Mandal's N.G.H Institute of Dental Sciences and Research Centre, Belgavi, Karnataka. MATERIALS AND METHODS: Fifty samples of each group were included in the present study. The levels of IL-21 were assessed using a commercially available enzyme-linked immunosorbent assay (ELISA) kit and the results were expressed as pg/mL. STATISTICAL ANALYSIS USED: Statistical analysis was performed using SPSS 17.0 software. Data were expressed as mean ± standard deviation and interquartile ranges and comparison of controls and cases by Mann-Whitney test. RESULTS: Serum and salivary levels of IL-21 were significantly higher in chronic periodontitis group than in controls (P < 0.001). Clinical periodontal parameters correlated positively with serum IL-21 levels. CONCLUSIONS: IL-21 is highly expressed in patients with chronic periodontitis and correlated well with clinical parameters of periodontal destruction. Therefore, IL-21 appears to play a role in tissue destruction and can be used as diagnostic biomarker in chronic periodontitis. Saliva can be considered to be a useful alternative to serum as a diagnostic sample.

Comparative evaluation of the efficacy of a herbal mouthwash and chlorhexidine mouthwash on select periodontal pathogens: An in vitro and ex vivo study.

BACKGROUND: Several herbal mouthwash and herbal extracts have been tested in vitro and in vivo in search of a suitable adjunct to mechanical therapy for long-term use. In this study, we aimed to look at the antimicrobial effect of the herbal mouthwash and chlorhexidine (CHX) mouthwash on select organisms in in vitro test and an ex vivo model. MATERIALS AND METHODS: The antimicrobial effects were determined against standard strains of bacteria that are involved in different stages of periodontal diseases. The in vitro tests included determination of minimum inhibitory concentration (MIC) using broth dilution and agar diffusion. In the ex vivo part of the study supragingival dental plaque were obtained from 20 periodontally healthy adult volunteers. Descriptive analysis was done for the entire quantitative and qualitative variable recorded. RESULTS: The MIC by broth dilution method found no statistically significant difference between the mouthwashes. The agar dilution method showed CHX was more effective as compared to the herbal mouthwash against standard strains of Streptococcus mutans, Streptococcus sanguinis, and Aggregatibacter actinomycetemcomitans. However, no difference was observed between the mouthwashes for Porphyromonas, Pseudomonas aeruginosa, and Fusobacterium nucleatum. The ex vivo results conclude that none of the selected mouthwashes were statistically significantly different from each other. CONCLUSION: In the present study, CHX showed higher levels of antimicrobial action than the herbal mouthwash against bacterial species. The results reinforce the earlier findings that the in vitro testing is sensitive to methods and due diligence is needed when extrapolating the data for further use. However, long-term use and in vivo effectiveness against the periopathogens need to be tested in well-planned clinical trials.

Detection and comparison of Selenomonas sputigena in subgingival biofilms in chronic and aggressive periodontitis patients.

BACKGROUND: With the advent of DNA-based culture-independent techniques, a constantly growing number of Selenomonas phylotypes have been detected in patients with destructive periodontal diseases. However, the prevalence levels that have been determined in different studies vary considerably. AIM: The present study was undertaken to detect and compare the presence of Selenomonas sputigena in the subgingival plaque samples from generalized aggressive periodontitis (GAP), chronic generalized periodontitis, and periodontally healthy patients using conventional polymerase chain reaction (PCR) technique. MATERIALS AND METHODS: A total of 90 patients were categorized as periodontally healthy individuals (Group I, n = 30), chronic generalized periodontitis (Group II, n = 30), and GAP (Group III, n = 30). The clinical parameters were recorded and subgingival plaque samples were collected. These were then subjected to conventional PCR analysis. STATISTICAL ANALYSIS USED: Kruskal-Wallis ANOVA test was used for multiple group comparisons followed by Mann-Whitney U-test for pairwise comparison. RESULTS: On comparison between three groups, all the clinical parameters were found to be statistically highly significant. Comparing Groups I-II and I-III, the difference in detection was found to be statistically highly significant whereas in Groups II-III, it was statistically nonsignificant. On comparison of S. sputigena detected and undetected patients to clinical parameters in various study groups, the difference was found to be nonsignificant. CONCLUSION: S. sputigena was found to be significantly associated with chronic and aggressive periodontitis. Although the difference in its detection frequency in both groups was statistically nonsignificant when compared clinically, S. sputigena was more closely associated with the GAP.

Occurrence of Aggregatibacter actinomycetemcomitans in Indian chronic periodontitis patients and periodontally healthy adults.

BACKGROUND: Aggregatibacter actinomycetemcomitans (Aa), an important primary periodontal pathogen, is known for its strong virulence characteristics that cause periodontal disease. We investigated Aa occurrence in Indian individuals using culture and 16 s rDNA polymerase chain reaction (PCR). MATERIALS AND METHODS: A cross-sectional study with 100 participants each in the healthy and chronic periodontitis (CP) groups was conducted. The subgingival plaque was collected and immediately plated on selective media for Aa. The remaining plaque samples were used for DNA extraction. PCR was performed using specific primers for Aa. STATISTICAL ANALYSIS USED: The detection of bacteria and the clinical parameters between the groups were compared using the Mann-Whitney U-test. For assessing the agreement between the results of anaerobic culture and PCR, Kappa analyses were performed. RESULTS: Aa levels using culture and PCR was 51% and 69% in the CP group and 12% and 30% in the healthy group, respectively. The two groups showed significant differences (P < 0.00001). The detection accuracy of culture and PCR was assessed, and the coefficient of accuracy (k) was highly significant in the healthy (0.3103; P < 0.0001) and CP groups (0.1536; P < 0.0497). CONCLUSIONS: Aa was predominantly found in the CP group compared with the healthy group, which is consistent with previous findings. Our results showed that both techniques can be used for detecting Aa. An ideal technique for detecting subgingival microorganisms should be carefully selected depending on the scope of the intended future work.

Prevalence of Herpesvirus and Correlation with Clinical Parameters in Indian Subjects with Chronic Periodontitis.

OBJECTIVE: The identification of new uncultured species and viruses supports the possibility of combination of the herpes-virus-bacterial periodontal infection for periodontitis. The paucity of data and studies with larger sample size in Indian subjects provides an unclear picture of the presence of the herpesvirus in this population. MATERIALS AND METHODS: This was a cross-sectional study consisting of 100 each in the healthy group and chronic periodontitis (CP) group. The subgingival plaque was collected and polymerase chain reaction was performed post deoxyribonucleic acid (DNA) extraction by using specific primers for human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV). The data were analyzed using Fisher's exact test, Mann-Whitney U test and Spearman's coefficient correlation. RESULTS: Human cytomegalovirus and EBV viruses were significantly higher in the CP group as compare to the healthy group. A higher percentage of those with CMV positive had EBV also positive (28.3%) compared to only 9.1% of CMV negative being EBV positive in the CP group. When both the healthy and CP group in total was compared, there was a significant correlation with all clinical parameters. CONCLUSION: Both the viruses dominated in disease as compared to health were similar to the earlier findings. The CP group had higher pocket depth and clinical attachment loss in the virus positive subjects. These findings could suggest that virus serves as a prelude to the disease and the combination of the two viruses could play a role in the pathogenesis.