Little Mahendra

16S rRNA-based detection of oral pathogens in coronary atherosclerotic plaque

BACKGROUND Atherosclerosis develops as a response of the vessel wall to injury. Chronic bacterial infections have been associated with an increased risk for atherosclerosis and coronary artery disease. The ability of oral pathogens to colonize in coronary atheromatous plaque is well known. AIM The aim of this study was to detect the presence of Treponema denticola, Porphyromonas gingivalis and Campylobacter rectus in the subgingival and atherosclerotic plaques of patients with coronary artery disease. MATERIALS AND METHODS Fifty-one patients in the age group of 40-80 years with coronary artery disease were selected for the study. DNA was extracted from the plaque samples. The specific primers for T. denticola, C. rectus and P. gingivalis were used to amplify a part of the 16S rRNA gene by polymerase chain reaction. STATISTICAL ANALYSIS USED Chi-square analysis, correlation coefficient and prevalence percentage of the microorganisms were carried out for the analysis. RESULTS Of the 51 patients, T. denticola, C. rectus and P. gingivalis were detected in 49.01%, 21.51% and 45.10% of the atherosclerotic plaque samples. CONCLUSIONS Our study revealed the presence of bacterial DNA of the oral pathogenic microorganisms in coronary atherosclerotic plaques. The presence of the bacterial DNA in the coronary atherosclerotic plaques in significant proportion may suggest the possible relationship between periodontal bacterial infection and genesis of coronary atherosclerosis.

Effect of curcumin against oxidation of biomolecules by hydroxyl radicals.

BACKGROUND Among various reactive oxygen species, hydroxyl radicals have the strongest chemical activity, which can damage a wide range of essential biomolecules such as lipids, proteins, and DNA. OBJECTIVE The objective of this study was to investigate the beneficial effects of curcumin on prevention of oxidative damage of biomolecules by hydroxyl radicals generated in in vitro by a Fenton like reaction. MATERIALS AND METHODS We have incubated the serum, plasma and whole blood with H2O2/Cu2+/ Ascorbic acid system for 4 hours at 37 0C and observed the oxidation of biomolecules like albumin, lipids, proteins and DNA. RESULTS Curcumin at the concentrations of 50,100 and 200 μmoles, prevented the formation of ischemia modified albumin, MDA, protein carbonyls, oxidized DNA and increased the total antioxidant levels and GSH significantly. CONCLUSION These observations suggest the hydroxyl radical scavenging potentials of curcumin and protective actions to prevent the oxidation of biomolecules by hydroxyl radicals.

Prevalence of eight putative periodontal pathogens in atherosclerotic plaque of coronary artery disease patients and comparing them with noncardiac subjects: A case-control study.

BACKGROUND The study was aimed to assess the prevalence of periodontal pathogens namely Tannerella forsythia (T.f), Campylobacter rectus (C.r), Eikenella corrodens (E.c), Porphyromonas gingivalis (P.g), Treponema denticola (T.d), Prevotella nigrescens (P.n) ,Aggregatibacter actinomycetemcomitans (A.a), P.g (fi mA gene) and Prevotella intermedia (P.i), in the subgingival and the atheromatous plaque of patients with coronary artery disease (CAD), and comparing them with the noncardiac subjects thereby indicating the role of periodontal pathogenic bacteria in the progression of atherosclerosis in south Indian population. MATERIALS AND METHODS 51 cardiac and non cardiac subjects within the age group of 40-80 years,who met the eligibility criteria, were selected and categorized as the experimental and control group respectively. Total number of teeth was recorded, and oral hygiene was evaluated using Plaque Index and Oral Hygiene Index (OHI). Periodontal pocket depth and clinical attachment level were also assessed as a part of periodontal examination. Subgingival plaque samples were collected with the help of with Graceys curette in both the groups. In experimental group, biopsy was obtained from the atherosclerotic plaque during Coronary artery bypass grafting CABG procedure. Both, subgingival and the coronary atherosclerotic plaque samples were subjected to polymerase chain reaction (PCR) analysis for identification of the periodontal bacteria. STATISTICAL ANALYSIS Mean, standard deviation and test of significance of quantitative variables such as periodontal parameters were compared between experimental group and control group. Kappa measures of agreement was done to analyze the relationship between the presence/absence of microorganisms in the subgingival and atherosclerotic plaque samples in the experimental group. RESULTS The mean Plaque Index, Gingival Index, Russels Periodontal Index, OHI, clinical attachment level, Pocket Depth Index was statistically significant in both the groups. Association of T.f, E.c, C.r, P.g, P.g (fi mA), T.d, P.i, P.n was significantly associated. A.a was absent in the control group, whereas only one patient in the experimental group was positive for the bacteria. Kappa analysis showed the significant association of periodontal bacteria T.f, C.r, P.g, P.g (fi mA), P.i and P.n in subgingival plaque and atherosclerotic plaque of the same patients of the experimental group. CONCLUSION It is concluded that CAD subjects had higher prevalence of periodontal pathogens in subgingival biofilms as compared to the non cardiac subjects. Further, the number of bacteria was significantly associated between the subgingival and atherosclerotic plaques of the cardiac patients in south Indian population.

Prevalence of periodontal pathogens in coronary atherosclerotic plaque of patients undergoing coronary artery bypass graft surgery

BackgroundChronic bacterial infections have been associated with an increased risk for atherosclerosis and coronary artery disease. The ability of oral pathogens to colonize in coronary atheromatous plaque is well known. The aim of our study was to detect the presence of four common periodontal pathogens in coronary plaques. We detected the presence of 16S rRNA of Treponema denticola, Eikenella Corrodens, Porphyromonas gingivalis and Campylobacter rectus in subgingival and atherosclerotic plaques of CABG surgery by using Polymerase Chain Reaction.Methods51 patients in the age group of 40 to 80 years with chronic periodontitis were recruited for the study. These patients were suffering from Coronary Artery Disease (CAD) and underwent Coronary Artery Bypass Grafting (CABG). DNA was extracted from the subgingival plaque and coronary atheromatous plaque samples. Universal Primer for the general detection of bacterial DNA and the primers for T.denticola, E. Corrodens, C.rectus and P.gingivalis were used to amplify part of 16SrRNA gene by Polymerase Chain Reaction.ResultsT.denticola, E.corrodens, C.rectus and P.gingivalis were detected in 49.01 %, 27.45 %, 21.51% and 45.10% of atherosclerotic plaque samples. In both subgingival and coronary plaque samples, T. denticola was detected in 39.21% of the cases, E.corrodens in 19.60%, C.rectus in 11.76% and P.gingivalis in 39.22% of the cases respectively.ConclusionOur study revealed the presence of significant bacterial DNA of oral pathogens in coronary plaques. This suggests possible relationship between periodontal infection and atherosclerosis and can help devise preventive treatment strategies.

Clinical effects of subgingivally delivered spirulina gel in chronic periodontitis cases: a placebo controlled clinical trial.

AIMS AND OBJECTIVES The aim of this study was to assess the clinical effects of Spirulina in-situ gel as an adjunct to Scaling And Root Planning (SRP) in the treatment of chronic periodontitis subjects. MATERIAL AND METHODS 64 sites were selected with probing pocket depth of ≥5mm and they were divided into 2 groups; 33 sites were treated with SRP along with spirulina gel (Group A) and 31 sites were treated with SRP alone (Group B). Clinical parameters were recorded at baseline before SRP and at 120(th) day after the treatment therapy. The parameters included Probing Pocket Depth (PPD) and Clinical Attachment Level (CAL). RESULTS Both the groups showed significant improvement in the parameters. However, Group A (SRP along with spirulina) showed statistically significant decrease in mean probing pocket depth and gain in the clinical attachment level after 120 days as compared to Group B SRP alone. CONCLUSION Locally delivered spirulina gel, along with scaling and root planning, has been shown to cause a beneficial impact. The efficacy of the product as a local drug delivery system in the non-surgical treatment of periodontitis without any side effects has been proved. Spirulina appears to be promising. It exerts strong anti-inflammatory effects which are closely connected with its antioxidative activity. This study can have a significant impact on the treatment procedures of periodontitis, with the use of blue green algae in the future.

Estimation of salivary neopterin in chronic periodontitis.

BACKGROUND Periodontal diseases are the most common bacterial infection predominantly associated with Gram-negative microorganisms that exist in the subgingival biofilm. Analysis of saliva provides a noninvasive means of evaluating the role of the host response in periodontal disease. Though salivary enzymes can be used as the biomarkers, neopterin has been recently used as one of the important diagnostic tools in the field of periodontics. Hence, we aimed to identify the neopterin levels in unsimulated saliva from the chronic periodontitis patients and compare them with the periodontally healthy subjects. MATERIALS AND METHODS Twenty subjects participated in the study and were categorized as the experimental group (chronic periodontitis patients) and control groups (healthy subjects). Unstimulated saliva samples were collected from both the groups for neopterin estimation. Neopterin in saliva was estimated using Shimadzu High Performance Liquid Chromatography with LC-20AD pump system, equipped with RF-10 AXL fluorescence detector. STATISTICAL ANALYSIS USED Data were expressed as mean±SD and analyzed using GraphPad Prism version 6.0 (California, USA). Statistical analysis was done by Students t-test. RESULTS The neopterin level in unstimulated saliva was found to be higher in the experimental group than the control group with P≤0.05. CONCLUSIONS The chronic periodontitis patients showed higher neopterin level in unstimulated saliva as compared to control. Hence, neopterin can be used as a potential biomarker for identification of the periodontal disease in its initial stage can help in preventing the disease progression.

Prevelance of periodontopathogenic bacteria in subgingival biofilm and atherosclerotic plaques of patients undergoing coronary revascularization surgery

Aim: The objective of the present study was to detect the presence of specific periodontopathogenic bacteria in the coronary plaque of patients with coronary artery disease and to find out the significant association between the periodontal status and the presence of pathogenic bacteria in the coronary plaque. Materials and Methods: The study population consisted of 51 patients with chronic generalized periodontitis undergoing coronary artery bypass grafting. Periodontal parameters were recorded and deoxyribonucleic acid was extracted from the subgingival plaque and coronary atherosclerotic plaque samples of the same patients. Polymerase chain reaction was used to amplify the part of 16S ribosomal ribonucleic acid (rRNA) gene to detect the presence of Aggregatibacter actinomycetemcomitans (Aa), Tannerella forsythia (Tf), Porphyromonas gingivali (Pg), Porphyromonas gingivalis (fimA) gene and Treponema denticola (Td). Results: Aa, Tf, Pg, Pg (fimA) gene and Td were detected in 0%, 31.4%, 45.1% 39.2% and 51% of atherosclerotic plaque samples, respectively. Tf was detected in 19.6%, Pg in 39.2%, Pg (fimA) gene in 33.3% and Td in 35.3% of both, subgingival plaque and atherosclerotic plaque samples. Periodontal parameters correlated with the presence of bacteria in coronary plaque. Aa could not be detected in coronary plaque samples. Conclusions: The study confirmed the detection of Red complex bacteria in coronary plaque samples and these bacteria correlated with the severity of periodontal destruction.

Clinical and microbiological efficacy of 4% Garcinia mangostana L. pericarp gel as local drug delivery in the treatment of chronic periodontitis: A randomized, controlled clinical trial

AIM The aim of the present study was to assess the periodontal status of individuals and the presence of red complex microorganisms (RCM), such as Treponema denticola (Td), Porphyromonas gingivalis, and Tannerella forsythia in the subgingival tissues of periodontitis patients before and after the application of 4% mangostana gel (Garcinia mangostana [MGA]) as an adjunct to scaling and root planing (SRP). METHODS Twenty-five patients (MGA group) were treated with SRP, and the subgingival application of mangostana gel was used as local drug delivery. Twenty-five patients (placebo group) were treated with SRP and placebo gel. Clinical parameters were recorded, and the presence of RCM was assessed at baseline and at the third month. RESULTS Clinical parameters, such as probing pocket depth, clinical attachment level, bleeding index, plaque index, and Td, were significantly reduced in the MGA group compared to the placebo group from baseline to the third month. CONCLUSION There was a significant improvement in the periodontal status with a reduction in Td with the application of mangostana gel in periodontal pockets. In the near future, 4% mangostana gel can be used as an adjunct to SRP to provide a new dimension to periodontal therapy.

Genetic analysis of Porphyromonas gingivalis (fimA), Aggregatibacter actinomycetemcomitans, and red complex in coronary plaque

AIM The objective of the present study was to detect the presence of Porphyromonas gingivalis (fimA), Aggregatibacter actinomycetemcomitans, and red complex in the coronary plaque of patients with coronary artery disease. METHODS The study population consisted of 51 patients with chronic periodontitis undergoing coronary artery bypass grafting. DNA was extracted from subgingival and coronary atherosclerotic plaque samples. Polymerase chain reaction was used to amplify the part of 16S rRNA gene to detect the presence of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis (fimA), Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola. RESULTS Aggregatibacter actinomycetemcomitans, Tannerella forsythia, Porphyromonas gingivalis, Porphyromonas gingivalis (fimA), and Treponema denticola were detected in 0%, 31.4%, 45.1%, 39.2%, and 51% of the atherosclerotic plaque samples, respectively. In both subgingival and coronary atherosclerotic plaque samples, Tannerella forsythia was detected in 19.6%, Porphyromonas gingivalis in 39.2%, Porphyromonas gingivalis (fimA) in 33.3%, and Treponema denticola in 35.3% of the samples. CONCLUSION The study confirmed the detection of red complex bacteria in coronary plaque samples. However Aggregatibacter actinomycetemcomitans could not be detected in these samples.

Evidence Linking the Role of Placental Expressions of Peroxisome Proliferator-Activated Receptor-γ and Nuclear Factor-Kappa B in the Pathogenesis of Preeclampsia Associated With Periodontitis

BACKGROUND Peroxisome proliferator-activated receptor (PPAR)-γ activation leads to suppression of production of a broad range of proinflammatory molecules. It plays a role in differentiation of trophoblasts and helps in normal placentation and formation of vascular exchange interface. Activation of nuclear factor-kappa (NF-κ) B triggers proinflammatory molecules inducing abnormal placentation and premature labor. This study aims to explore expression of PPAR-γ and NF-κB in placentas of women with periodontitis-associated preeclampsia compared with that in normotensive pregnant women. METHODS Fifty pregnant women were included. Twenty-five were controls (normotensive pregnant women) and 25 were pregnant women with preeclampsia, including those with gestational hypertension. Demographic data, pregnancy characteristics, and periodontal parameters were recorded, including: 1) plaque index; 2) gingival index; 3) bleeding on probing (BOP); 4) probing depth; and 5) attachment loss (AL). Placental tissue samples were collected from both groups and analyzed to quantify expression of PPAR-γ and NF-κB using real-time polymerase chain reaction. RESULTS BOP and AL were significantly higher in pregnant women with preeclampsia compared with normotensive pregnant women (P <0.05). Expression of PPAR-γ was downregulated in patients with preeclampsia compared with that of healthy normotensive patients, which was statistically significant (P <0.05), whereas NF-κB was significantly activated (P <0.05) in pregnant women with preeclampsia compared with normotensive pregnant women. CONCLUSIONS Higher periodontal disease prevalence is found among pregnant women with preeclampsia, with increased percentage of sites with BOP and greater AL. This study provides novel information on host response to systemic inflammation induced by periodontal pathogens through mechanisms involving downregulation of PPAR-γ and increased activation of NF-κB.