By using NGS-sequencing libraries of DNA from periodontal swabs with primers specific to V6 region of 16S rDNA prevalence of bacterial genera and species in periodontal microbiota of patients with aggressive periodontitis and healthy donors was analyzed. Six genera of putative periodontal protectors and eight periodontal pathogens were identified with respect to aggressive (but not chronic) periodontitis. Statistically relevant over-colonization by general Porphyromonas, Treponema, Synergistes, Tannerella, Filifactor, Ruminococcus, Parvimonas and Mycoplasma was found to be associated with the condition. From these, only three genera Porphyromonas, Treponema and Tannerella are traditionally considered as periodontal pathogens. Statistically confidential over-colonization by genus Veillonella was found in healthy patients. This genus should be considered as a relevant marker of a healthy periodontium. Genera Streptococcus, Bergeyella, Granulicatella, Kingella and Corynebacterium may be considered as putative periodontal protectors. Comparison of data of NGS-sequencing and real-time PCR demonstrated a good agreement if different PCR efficiency using independent primer pairs is taken into account.
By using qPCR system, women as well as men were found to have an equal periodontal pathogen colonization. However, the women are subjected to have a higher risk of chronic periodontitis onsets. Women with the chronic periodontitis usually expose an evident hypercolonization with a single pathogen. P. gingivalis is the most prevalent causative agent of the chronic periodontitis in women but not in men. In health as well as in the chronic periodontitis a complex of periodontal pathogens forms such as P. gingivalis, P. intermedia, T. forsythensis and T. denticola. T. forsythensis demonstrates the highest correlation with the chronic periodontitis onset in men. Our data allow us to prove T. forsythensis playing the key role in the forming of periodontal pathogen complex.
The study revealed positive correlation between bleeding on probing and teeth loss risk with periodontal hypercolonization by Porphyromonas gingivalis, Prevotella intermedia and Treponema denticola. Pathological tooth mobility was associated with hypercolonization by P. intermedia and Tannerella forsythensis. Expression of IL8, TNF-α, MMP8 and MMP9 genes was also assessed in patient groups divided according to the depth of periodontal pockets and-the severity of chronic periodontitis revealing IL8 as positive diagnostic marker.
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