Correlation Between Intestinal Clostridium Leptum And Periodontal Health

Background:Periodontitis is a common oral chronic infectious disease whose development and progression is affected by many factors. Epidemiological studies reveal that more than two-thirds of the world’s population suffers from one of the chronic forms of periodontal disease, as well as forty-four percent to fifty-seven percent and ten percent of adults suffer from moderate periodontitis and severe periodontitis, respectively. Although periodontal pathogens are initiating factor for the initiation of periodontitis, the disease process is ultimately decided by the host immune responses. Regulatory T cells (Treg) as the main immune suppression and regulation cells play an irreplaceable role in the development and progression of periodontitis.Type2diabetes mellitus is a common endocrine metabolic disease, which is considered to be one of the risk factors for periodontal disease. The clinical study results indicate that the incidence and severity of periodontitis in type2diabetes mellitus patients were higher than those without diabetes in the case of local incentives similar. A large number of studies suggest that type2diabetes mellitus is a low-grade chronic inflammation related to immune dysfunction. With the acceleration of global changes in lifestyle and the aging process, the prevalence of periodontitis and type2diabetes are rapidly increasing and they has become a worldwide public health problem. Therefore, investigating the new method of preventing this class of inflammatory diseases has important scientific significance and far-reaching social significance.In recent years, several studies have demonstrated that indigenous species of Clostridium leptum subgroup (C. leptum) in the gut can promote the generation of Treg, and suppress self-injury inflammatory response by expanding and activating Treg as well as secreting anti-inflammatory cytokines (such as IL-10), thereby reducing the body’s immune pathological damage. A recent research found that oral feeding with C. leptum can effectively induce Treg proliferation and activation in adult mice and significantly reduce inflammatory cells infiltration and inflammatory cytokine secretion in OVA sensitized mice airway. These findings suggest that C. leptum may involve in inflammatory diseases and reduce the body’s inflammatory response to a certain extent, and this will produce important guidance significance to scientific research and clinical treatment of periodontitis.Although the role of C. leptum in inflammatory diseases has drawn increasingly concern of experts and scholars at home and abroad, researches about C. leptum and periodontal tissue inflammation have not been reported. Up to now, it is not clear whether C. leptum is associated with periodontal inflammatory status, or whether it plays an anti-inflammatory role to reduce the inflammatory destruction of periodontal tissue.Objective:The purpose of this study was to preliminary investigate the relationship between C. leptum and inflammation of periodontal tissue in patients with chronic periodontitis and type2diabetes mellitus, as well as to provide new ideas for the clinical treatment of periodontitis by using real-time quantitative polymerase chain reaction (RTQ-PCR) quantitatively detected C. leptum.Methods:Fresh fecal samples were obtained from twenty patients with chronic periodontitis (CP group), twenty patients with type2diabetes mellitus (DM group), twenty patients with chronic periodontitis accompanying type2diabetes [(DM+CP) group] and twenty healthy control subjects (HC group), and each subject was conducted with a questionnaire survey and periodontal clinical examination. Genus-specific primer of C. leptum subgroup was designed for16S rRNA-targeted PCR. The standard curve of real-time quantitative polymerase chain reaction (RTQ-PCR) was generated from a grade-dilution of plasmid DNA which was cloned from PCR products. Bacterial genome DNA extracted from fecal samples was quantified by RTQ-PCR to analyze C. leptum amounts. SPSS17.0software was used for statistical analysis. Measurement data was expressed as mean±standard deviation, PD, AL and bacterial gene copy numbers in each group was used by compare t-test, BOP positive rate was used by chi-square test, P<0.05was considered statistically significant.Results:1. PD, AL and BOP positive rates comparison:PD and AL of (DM+CP) group were significantly greater than HC group, CP group and DM group (P<0.05). BOP positive rates of (DM+CP) group were significantly higher than HC group, CP group and DM group(P<0.05).2. Logarithm for16S rRNA gene copies of C. leptum per gram feces in HC group, CP group, DM group and (DM+CP) group were (7.09±0.17),(6.31±0.58),(5.79±0.52) and (5.41+0.29), respectively. Compared with HC group, the gene copies of C. leptum in CP group, DM group and (DM+CP) group were decreased dramatically (P<0.01). The gene copies of C. leptum in (DM+CP) group were significantly lower than CP group and DM group, P<0.01was considered statistically significant.Conclusion:The amount of intestinal flora C. leptum is associated with periodontal tissue inflammatory state. Compared with HC group, CP group and DM group,(DM+CP) group which has the least number of C. leptum having the most severely periodontal tissue destruction. It suggests that low levels of C. leptum may not be sufficient to inhibit excessive self-injury immune response, thereby aggravate local inflammation of the periodontal tissue.