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A Microbiological Profile of Unexposed and Exposed Pulp Space of Primary Endodontic Infections by Checkerboard DNA-DNA Hybridization

      Abstract

      Introduction

      The aim of this study was to evaluate, by checkerboard DNA-DNA hybridization, the composition of the microbiota of primary endodontic infections in cases associated with exposed (n = 30) and unexposed (n = 30) pulp space.

      Methods

      Samples were collected by means of a #15 H-type file and 2 sterile paper points from 60 single-rooted teeth with necrotic pulp and periapical lesions. The presence, levels, and proportions of 40 bacterial species were determined by checkerboard DNA-DNA hybridization.

      Results

      The species found in higher counts (×105) in exposed pulp space cases were Eubacterium saburreum, Fusobacterium nucleatum ssp. vincentii, Tannerella forsythia, Enterococcus faecalis, Neisseria mucosa, Campylobacter gracilis, and Prevotella nigrescens, and in unexposed pulp space cases they were F. nucleatum ssp. vincentii, N. mucosa, E. faecalis, E. saburreum, C. gracilis, and Porphyromonas gingivalis. Counts of F. nucleatum ssp. vincentii, Campylobacter sputigena, Capnocytophaga showae, Treponema socrenskii, Porphyromonas endodontalis, Eikenella corrodens, and Capnocytophaga ochracea were significantly higher in unexposed pulp space cases (P < .05).

      Conclusions

      The data of the present investigation suggested specific differences between the composition of the microbiota in cases with exposed and unexposed pulp space and an association between higher levels of some specific species and unexposed pulp space cases.

      Key Words

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      References

        • Miller W.D.
        An introduction to the study of the bacterio-pathology of the dental pulp.
        Dental Cosmos. 1894; 36: 505-528
        • Kakehashi S.
        • Stanley H.R.
        • Fitzgerald R.J.
        The effects of surgical exposures of dental pulps in germ-free and conventional laboratory rats.
        Oral Surg Oral Med Oral Pathol. 1965; 20: 340-349
        • Sundqvist G.
        Bacteriological Studies of Necrotic Dental Pulps (PhD Thesis).
        University of Umea, Umea, Sweden1976
        • Sundqvist G.
        • Johansson E.
        • Sjögren U.
        Prevalence of black-pigmented bacteroides species in root canal infections.
        J Endod. 1989; 15: 13-19
        • Baumgartner J.C.
        • Falkler W.A.
        Bacteria in the apical 5 mm of infected root canals.
        J Endod. 1991; 17: 380-383
        • Gomes B.P.F.A.
        • Lilley J.D.
        • Drucker D.B.
        Associations of endodontic symptoms and signs with particular combinations of specific bacteria.
        Int Endod J. 1996; 29: 69-75
        • Jung I.Y.
        • Choi B.K.
        • Kum K.Y.
        • et al.
        Identification of oral spirochetes at the species level and their association with other bacteria in endodontic infections.
        Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2001; 92: 329-334
        • Siqueira Jr., J.F.
        • Rôças I.N.
        • Souto R.
        • Uzeda M.
        • Colombo A.P.
        Checkerboard DNA-DNA hybridization analysis of endodontic infections.
        Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2000; 89: 744-748
        • Siqueira Jr., J.F.
        • Rôças I.N.
        • Favieri A.
        • Oliveira J.C.M.
        • Santos K.R.N.
        Polymerase chain reaction detection of Treponema denticola in endodontic infections within root canal.
        Int Endod J. 2001; 34: 280-284
        • Gomes B.P.F.A.
        • Jacinto R.C.
        • Pinheiro E.T.
        • et al.
        Molecular analysis of Filifactor alocis, Tannerella forsythia and Treponema denticola associated with primary endodontic infections and failed endodontic treatment.
        J Endod. 2006; 32: 937-940
        • Siqueira Jr., J.F.
        • Rôças I.N.
        Treponema socranskii in primary endodontic infections as detected by Nested PCR.
        J Endod. 2003; 29: 244-247
        • Sassone L.
        • Fidel R.
        • Figueiredo L.
        • Fidel S.
        • Faveri M.
        • Feres M.
        Evaluation of the microbiota of primary endodontic infections using checkerboard DNA-DNA hybridization.
        Oral Microbiol Immunol. 2007; 22: 390-397
        • Sassone L.
        • Fidel R.
        • Faveri M.
        • et al.
        A microbiological profile of symptomatic teeth with primary endodontic infections.
        J Endod. 2008; 34: 541-545
        • Siqueira Jr., J.F.
        • Rôças I.N.
        Bacteroides forsythus in primary endodontic infections as detected by Nested PCR.
        J Endod. 2003; 29: 390-393
        • Souza C.A.S.
        • Teles R.P.
        • Souto R.
        • Chaves M.A.E.
        • Colombo A.P.
        Endodontic therapy associated with calcium hydroxide as na intracanal dressing: microbiological evaluation by the checkerboard DNA-DNA hybridization technique.
        J Endod. 2005; 31: 79-83
        • Rôças I.N.
        • Alves F.R.F.
        • Santos A.L.
        • Rosado A.S.
        • Siqueira Jr., J.F.
        Apical root canal microbiota as determined by reverse-capture checkerboard analysis of cryogenically ground root samples from teeth with apical periodontitis.
        J Endod. 2010; 36: 1617-1621
        • Sedgley C.
        • Nagel A.
        • Dahlén G.
        • Reit C.
        • Molander A.
        Real-time quantitative polymerase chain reaction and culture analyses of Enterococcus faecalis in root canals.
        J Endod. 2006; 32: 173-177
        • Siqueira Jr., J.F.
        • Rôças I.N.
        Detection of Filifactor alocis in endodontic infections associated with different forms of perirradicular diseases.
        Oral Microbiol Immunol. 2003; 18: 263-265
        • Socransky S.S.
        • Smith C.
        • Martin L.
        • Paster B.J.
        • Dewhirst F.E.
        • Levin A.E.
        “Checkerboard” DNA-DNA hybridization.
        Biotechniques. 1994; 17: 788-792
        • Sundqvist G.
        Taxonomy, ecology, and pathogenicity of root canal flora.
        Oral Surg Oral Med Oral Pathol. 1994; 78: 522-530
        • Sundqvist G.
        Ecology of the root canal flora.
        J Endod. 1992; 18: 427-430
        • Chu F.C.S.
        • Tsang C.S.P.
        • Chow T.W.
        • Samaranayake L.P.
        Identification of cultivable microorganisms from primary endodontic infections with exposed and unexposed pulp space.
        J Endod. 2005; 31: 424-429
        • Haffajee A.D.
        • Cugini M.A.
        • Dibart S.
        • Smith C.
        • Kent Jr., R.L.
        • Socransky S.S.
        Clinical and microbiological features of subjects with adult periodontitis who responded poorly to scaling and root planning.
        J Clin Periodontol. 1997; 24: 767-776
        • Feinberg A.P.
        • Vogelstein B.
        A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.
        Anal Biochem. 1983; 132: 6-13
        • Sakamoto M.
        • Rôças I.N.
        • Siqueira Jr., J.F.
        • Benno Y.
        Molecular analysis of bacteria in asymptomatic and symptomatic endodontic infections.
        Oral Microbiol Immunol. 2006; 21: 112-122
        • Siqueira Jr., J.F.
        • Rôças I.N.
        Diversity of endodontic microbiota revisited.
        J Dent Res. 2009; 88: 969-981
        • Ribeiro A.C.
        • Matarazzo F.
        • Faveri M.
        • Zezell D.M.
        • Mayer M.P.A.
        Exploring bacterial diversity of endodontic microbiota by cloning and sequencing 16S rRNA.
        J Endod. 2011; 37: 922-926
        • Montagner F.
        • Gomes B.P.F.A.
        • Kumar P.S.
        Molecular fingerprinting reveals the presence of unique communities associated with paired samples of root canals and acute apical abscesses.
        J Endod. 2010; 36: 1475-1479
        • Rôças I.N.
        • Siqueira Jr., J.F.
        • Debelian G.J.
        Analysis of symptomatic and asymptomatic primary root canal infections in adult Norwegian patients.
        J Endod. 2011; 37: 1206-1212
        • Brown Jr., L.R.
        • Rodolph Jr., C.E.
        Isolation and identification of microorganism from exposed canals of pulp-involved teeth.
        Oral Surg Oral Med Oral Pathol. 1957; 10: 1094-1099
        • Siqueira Jr., J.F.
        • Rôças I.N.
        • Rosado A.S.
        Investigation of bacterial communities associated with asymptomatic and symptomatic endodontic infections by denaturing gradient gel electrophoresis fingerprinting approach.
        Oral Microbiol Immunol. 2004; 19: 363-370
        • Siqueira Jr., J.F.
        • Rôças I.N.
        Exploiting molecular methods to explore endodontic infections: part 1—current molecular technologies for microbial diagnosis.
        J Endod. 2005; 31: 411-423