Subgingival microbiome in smokers and non-smokers in periodontitis: an exploratory study using traditional targeted techniques and a next-generation sequencing

Authors

  • Sergio Bizzarro,

    Corresponding author
    • Department of Periodontology, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Amsterdam, The Netherlands
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  • Bruno G. Loos,

    1. Department of Periodontology, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Amsterdam, The Netherlands
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  • Marja L. Laine,

    1. Department of Periodontology, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Amsterdam, The Netherlands
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  • Wim Crielaard,

    1. Department of Preventive Dentistry, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Amsterdam, The Netherlands
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  • Egija Zaura

    1. Department of Preventive Dentistry, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Amsterdam, The Netherlands
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  • Conflict of interest and source of funding statement

    The authors declare that they have no conflict of interest.

    This study was funded in part by the authors' institution and in part by a grant from the University of Amsterdam for the focal point “Oral infection and inflammation”.

Address:

Sergio Bizzarro

Department of Periodontology

Academic Centre for Dentistry Amsterdam (ACTA)

Gustav Mahlerlaan 3004

1081 LA, Amsterdam, The Netherlands

E-mail: s.bizzarro@acta.nl

Abstract

Aim

To compare the results of two targeted techniques to an open-ended technique in periodontitis patients, differentiated on the basis of smoking habit.

Materials & Methods

Thirty periodontitis patients (15 smokers and 15 non-smokers) provided subgingival plaque samples for 16S rRNA gene amplicon sequencing, culturing and quantitative polymerase chain reaction (qPCR).

Results

No differences were found in the composition of the subgingival microbiome between smokers and non-smokers with culture and qPCR. With pyrosequencing, operational taxonomic units (OTUs) classified to genera Fusobacterium, Prevotella and Selenomonas were more abundant in smokers, while OTUs belonging to the genera Peptococcus and Capnocytophaga were more abundant in non-smokers. Principal coordinate analysis identified two clusters; one was composed mainly of smokers (80%) and revealed significantly lower taxonomic diversity, higher attachment loss and higher proportion of the genera Fusobacterium, Paludibacter and Desulfobubus.

Conclusion

In periodontitis, there is a difference in the composition of the subgingival microbiome between smokers and non-smokers, as revealed by pyrosequencing. This difference was not identified by the targeted techniques. Low taxonomic diversity was associated with higher disease severity, especially in smokers. This supports the hypothesis of the ecological microbial–host interaction in the severity of periodontal disease.

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