title: Molecular evaluation of residual endodontic microorganisms after instrumentation, irrigation and medication with either calcium hydroxide or Septomixine.
abstract: OBJECTIVE
  The correct choice of antimicrobial agents as inter-appointment medicaments is as important as the instrumentation and irrigation to remove pathogens from infected root canals. Calcium hydroxide [Ca(OH)2] and framycetin sulfate (Septomixine) are common endodontic medicaments. Therefore, we evaluated the efficacy of either calcium hydroxide or Septomixine in eliminating residual intra-canal bacteria, particularly Actinomyces spp., during inter-appointment interval in endodontic therapy using molecular methods.
  
  
  METHODS
  A total of 31 single-rooted teeth with primary root canal infections were studied immediately after opening the canals and subsequently after instrumentation, irrigation with sterile saline and 1-week medication with either Ca(OH)2 (n = 25) or Septomixine (n = 6). Whole bacterial genomic DNA was isolated directly from samples and PCR with universal primers performed to detect total intra-canal bacteria. The variable regions of 16S rDNA of bacteria were amplified and labeled with digoxigenin for further hybridization to detect Actinomyces spp. A total of seven oligonucleotide probes specific for A. bovis, A. gerencseriae, A. israelii, A. meyeri, catalase-negative A. naeslundii (genospecies 1 and 2), catalase-positive A. naeslundii genospecies 2 and A. odontolyticus were used to detect Actinomyces spp. in 22 of 31 medicated root canals [Ca(OH)2: n = 17; Septomixine: n = 5].
  
  
  RESULTS
  The PCR results showed that 25 of 31 examined canals were positively detected with residual microorganisms after instrumentation, irrigation with sterile saline and 1-week medication with either Ca(OH)2 (n = 20) or Septomixine (n = 5). Thus, only six canals [Ca(OH)2: n = 5, Septomixine: n = 1] were aseptic after treatment. Hybridization results showed higher detection frequency of both A. odontolyticus and A. gerencseriae after treatment. Significant correlation was found between exposed pulp before treatment and positive detection of Actinomyces spp., particularly A. odontolyticus on the second visit (P < 0.05).
  
  
  CONCLUSIONS
  The conventional, 1-week medication of either Ca(OH)2 or Septomixine in endodontic therapy may not effectively inhibit residual bacterial growth in all root canals during inter-appointment intervals. Further investigations using, for instance quantitative real-time PCR analyses, are required to substantiate the present findings.
authors.affiliation: Oral Bio-Sciences, Faculty of Dentistry, The University of Hong Kong, Hong Kong.
authors.forename: G
authors.initials: G
authors.lastname: Tang
authors.forename: L P
authors.initials: LP
authors.lastname: Samaranayake
authors.forename: H-K
authors.initials: HK
authors.lastname: Yip
mesh_terms: D000196:Actinomycosis / Q000188:drug therapy* / Q000382:microbiology; D000293:Adolescent; D000328:Adult; D000368:Aged; D000900:Anti-Bacterial Agents / Q000627:therapeutic use*; D002126:Calcium Hydroxide / Q000627:therapeutic use*; D003786:Dental Pulp Cavity / Q000187:drug effects / Q000382:microbiology*; D005260:Female; D005601:Framycetin / Q000627:therapeutic use*; D006801:Humans; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods; D016017:Odds Ratio; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction
chemical_list: D000900:Anti-Bacterial Agents; D015345:Oligonucleotide Probes; D005601:Framycetin; D002126:Calcium Hydroxide