Shift of microbial composition of peri-implantitis-associated oral biofilm as revealed by 16S rRNA gene cloning

J Med Microbiol. 2018 Mar;67(3):332-340. doi: 10.1099/jmm.0.000682. Epub 2018 Jan 24.

Abstract

Purpose: Micro-organisms are important triggers of peri-implant inflammation and analysing their diversity is necessary for peri-implantitis treatment. This study aimed to analyse and compare the microbiota associated with individuals with peri-implantitis, as well as clinically healthy implant sites.

Methodology: Subgingival biofilm samples were taken from 10 individuals with peri-implantitis and from at least 1 clinically healthy implant. DNA was extracted and bacterial 16S rRNA genes were amplified using universal primers. After cloning the PCR-products, amplified inserts of positive clones were digested using restriction endonucleases, and the chosen clones were sequenced. The 16S rDNA-sequences were compared to those from the public sequence databases GenBank, EMBL and DDBJ to determine the corresponding taxa.

Results: Differing distributions of taxa belonging to the phyla Firmicutes, Bacteroidetes, Fusobacteria, Actinobacteria, Proteobacteria, Synergistetes, Spirochaetae and TM 7 were detected in both the healthy implant (HI) and the peri-implantitis (PI) groups. A significantly higher relative abundance of phylum Bacteroidetes, as well as of the species Fusobacterium nucleatum, were found in the PI group (P<0.05). The putative periodontal red complex (Porphyromonas gingivalis, Tannerella forsythia) was also detected at significantly higher levels in the PI group (P<0.05), whereas the yellow group, as well as the species Veillonella dispar, tended to be associated with the HI group.

Conclusion: A shift in the healthy subgingival microbiota was shown in peri-implantitis-associated biofilm. Anaerobic Gram-negative periopathogens, including P. gingivalis and T. forsythia, seem to play an important role in peri-implantitis development and should be considered in treatment and prevention strategies.

Keywords: culture-independent method; microbiota; peri-implant disease; subgingival biofilm; subgingival microbial complexes.

MeSH terms

  • Aged
  • Aged, 80 and over
  • Bacteria / classification
  • Bacteria / genetics
  • Bacteria / isolation & purification*
  • Bacterial Load
  • Bacterial Physiological Phenomena
  • Bacteroides / genetics
  • Bacteroides / isolation & purification
  • Biofilms*
  • Female
  • Fusobacterium nucleatum / genetics
  • Fusobacterium nucleatum / isolation & purification
  • Genes, rRNA
  • Gingiva / microbiology
  • Humans
  • Male
  • Microbiota / genetics*
  • Middle Aged
  • Peri-Implantitis / microbiology*
  • Porphyromonas gingivalis / genetics
  • Porphyromonas gingivalis / isolation & purification
  • Prevotella intermedia / genetics
  • Prevotella intermedia / isolation & purification
  • RNA, Ribosomal, 16S / genetics*
  • Sequence Analysis, DNA

Substances

  • RNA, Ribosomal, 16S