IADR Abstract Archives
Smoking, Diabetes, and the Subgingival Microbiome
Objectives: Smoking and diabetes are known risk factors for periodontitis, and their effect on the subgingival biofilm have been independently studied. However the combined effect of these periodontitis risk factors is unknown. The aim of the current study is to analyze and compare the microbial composition of the subgingival biofilm in diabetics, smokers, and diabetic smokers with periodontal disease.
Methods: The study population consisted of 100 periodontally diseased subjects divided evenly between four groups classified based on systemic health status: diabetics, smokers, diabetic smokers, and non-diabetic, non-smoking controls. Subgingival biofilm samples were collected from shallow (pockets <3mm) and deep (pockets >3mm) sites using endodontic paper points. DNA was isolated with Qiagen MiniAmp kit according to the manufacturer’s instructions. Amplification of the bacterial 16S rRNA gene was accomplished by PCR using universal primers targeting the V1-V3 and V7-V9 hypervariable regions. 454 pyrosequencing was performed, and the generated sequences were compared to the Human Oral Microbiome Database. Statistical comparisons were made using Tukey HSD.
Results: In all four groups there were no differences in microbial profiles between deep and shallow sites. Principal coordinate analysis of Unifrac distances revealed the microbial composition of smokers and diabetics were similar, however smokers demonstrated a more homogenous clustering than diabetics. Both smokers and diabetics were distinctly different from diabetic smokers. This was apparent in deep and shallow sites. The species contributing to these differences included Porphyomonas gingivalis, Streptococcus mutans, Capnocytophaga gingivalis, Campylobacter gracilis, Fretibacterium fastidiosum, Fusobacterium nucleatum, Streptococcus sanguinis, Veillonella atypical, Gemella haemolysans, and Treponema parvum.
Conclusions: Smoking and diabetes have distinct effects on the microbiome. The heterogenous microbial composition in diabetic smokers suggests a synergistic effect of these periodontitis risk factors on the subgingival microbiome. Shallow sites are at-risk-for-harm, therefore clinical probing depths are not indicative of a more health-associated subgingival microbiome.
Methods: The study population consisted of 100 periodontally diseased subjects divided evenly between four groups classified based on systemic health status: diabetics, smokers, diabetic smokers, and non-diabetic, non-smoking controls. Subgingival biofilm samples were collected from shallow (pockets <3mm) and deep (pockets >3mm) sites using endodontic paper points. DNA was isolated with Qiagen MiniAmp kit according to the manufacturer’s instructions. Amplification of the bacterial 16S rRNA gene was accomplished by PCR using universal primers targeting the V1-V3 and V7-V9 hypervariable regions. 454 pyrosequencing was performed, and the generated sequences were compared to the Human Oral Microbiome Database. Statistical comparisons were made using Tukey HSD.
Results: In all four groups there were no differences in microbial profiles between deep and shallow sites. Principal coordinate analysis of Unifrac distances revealed the microbial composition of smokers and diabetics were similar, however smokers demonstrated a more homogenous clustering than diabetics. Both smokers and diabetics were distinctly different from diabetic smokers. This was apparent in deep and shallow sites. The species contributing to these differences included Porphyomonas gingivalis, Streptococcus mutans, Capnocytophaga gingivalis, Campylobacter gracilis, Fretibacterium fastidiosum, Fusobacterium nucleatum, Streptococcus sanguinis, Veillonella atypical, Gemella haemolysans, and Treponema parvum.
Conclusions: Smoking and diabetes have distinct effects on the microbiome. The heterogenous microbial composition in diabetic smokers suggests a synergistic effect of these periodontitis risk factors on the subgingival microbiome. Shallow sites are at-risk-for-harm, therefore clinical probing depths are not indicative of a more health-associated subgingival microbiome.