title: Identification of Actinobacillus actinomycetemcomitans, Treponema denticola and Porphyromonas gingivalis within human dental calculus: a pilot investigation.
abstract: OBJECTIVE
  Dental calculus is considered to be simply a "plaque-retentive factor", and therefore only a secondary aetiological factor in the pathogenesis of periodontitis. Recent studies have suggested a more active role for calculus. Our objective was to demonstrate the presence of periodontal pathogens in the non-mineralised areas of supra- and subgingival dental calculus.
  
  
  METHODS
  Subjects for the study were derived from patients with substantial amounts of supragingival calculus in the lower anterior region who had moderate periodontal disease, having been referred to the periodontal department at the Eastman Dental Hospital for periodontal care. Calculus was removed in as large pieces as possible by the use of a sickle or a push scaler placed underneath the apical or facial border of the calculus and fracturing it from the tooth surface in a single stroke. The orientation and absence of dental plaque was confirmed using light microscopy for each sample prior to inclusion in this study. Samples were prepared for transmission electron microscopic (TEM) observation after immunogold staining with polyclonal antibodies for the presence of Actinobacillus actinomycetemcomitans (A. a.), Porphyromonas gingivalis (P. g.) and Treponema denticola (T. d.).
  
  
  RESULTS
  Most of the samples contained at least one of the bacterial species examined, either in the lacunae or in the covering dental plaque. T. d. was the most frequently identified species and was found in nearly all of the subgingival samples, whilstA. a. was rarely observed.
  
  
  CONCLUSIONS
  In this limited study, supra- and subgingival dental calculus appears to be capable of maintaining periodontal pathogens within the deep recesses of its structural lacunae and channels. Therefore, calculus could possibly play a relevant role in the aetiology and pathogenesis of periodontitis. The presence of T. d. in the majority of specimens requires further investigation as its pathogenic potential may be underestimated in current published microbiological research, and further work is required to determine its role in the pathogenesis of periodontal diseases. However, further work by way of a large-scale definitive study is necessary to confirm the results of this investigation.
authors.affiliation: Eastman Dental Institute for Oral Health Care Sciences, UCL, 256 Grays Inn Road, London WC1X 8LD, United Kingdom.
authors.forename: Nicolino
authors.initials: N
authors.lastname: Calabrese
authors.forename: Peter
authors.initials: P
authors.lastname: Galgut
authors.forename: Nicola
authors.initials: N
authors.lastname: Mordan
mesh_terms: D000328:Adult; D016976:Aggregatibacter actinomycetemcomitans / Q000302:isolation & purification*; D000907:Antibodies, Bacterial; D015331:Cohort Studies; D015169:Colony Count, Microbial; D003728:Dental Calculus / Q000382:microbiology*; D003773:Dental Plaque / Q000382:microbiology; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D046529:Microscopy, Electron, Transmission; D008875:Middle Aged; D010518:Periodontitis / Q000382:microbiology; D010865:Pilot Projects; D016966:Porphyromonas gingivalis / Q000302:isolation & purification*; D048373:Treponema denticola / Q000302:isolation & purification*
chemical_list: D000907:Antibodies, Bacterial