Comparison of Efficacy of Sterile Paper Point, Dentin Harvesting and Pulverization Techniques for Sampling Root Canals using Real Time-PCR: An in Vitro Study
Keywords:
sampling, pulverization, dentin harvesting, PCR, paper pointAbstract
Endodontic infections, primarily caused by biofilm-forming bacteria such as Enterococcus faecalis and Streptococcus mutans, pose significant challenges due to their resistance to treatment and host defenses. This study evaluated three sampling techniques—Pulverization, Dentin Harvesting, and Sterile Paper Points—for bacterial recovery and molecular quantification in endodontic research. Sixty sterilized single-rooted human teeth were inoculated with E. faecalis and S. mutans to simulate polymicrobial infections. Bacterial recovery was assessed using colony-forming unit (CFU) counts, while DNA quantification was performed via quantitative PCR (qPCR). Pulverization demonstrated the highest bacterial recovery and DNA yield, attributed to its effective disruption of dentinal biofilms. Dentin Harvesting showed moderate recovery and was valuable for studying microbial distribution within dentin layers. Sterile Paper Points yielded the lowest recovery due to their inability to disrupt biofilms or access bacteria embedded in dentinal tubules. qPCR analysis corroborated these results, with Pulverization being the most efficient technique for both total bacterial and species-specific quantification. These findings emphasize Pulverization as the most effective method for comprehensive microbial analysis, while Dentin Harvesting and Sterile Paper Points offer targeted or surface-level applications. This study highlights the importance of selecting appropriate sampling methods to enhance the accuracy of microbial and molecular analyses in endodontic infections.
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Baumgartner JC, Siqueira JF Jr, Sedgley CM, Kishen A. Microbiology of endodontic disease. Endodontic Topics. 2003;7(1):42-76.
Mohammadi Z, Shalavi S, Yazdizadeh M, Jafarzadeh H. Antimicrobial effects of calcium hydroxide in endodontics: a review. Chonnam Med J. 2012;48(3):133-140.
Love RM. Enterococcus faecalis—a mechanism for its role in endodontic failure. Int Endod J. 2001;34(5):399-405.
Stuart CH, Schwartz SA, Beeson TJ, Owatz CB. Enterococcus faecalis: its role in root canal treatment failure and current concepts in retreatment. J Endod. 2006;32(2):93-98.
Chávez de Paz LE. Redefining the persistent infection in root canals: possible role of biofilm communities. J Endod. 2007;33(6):652-662.
de Souza EL, Meira QG, Barbosa I, et al. The resistance of Enterococcus faecalis biofilm to different intracanal medications. J Appl Oral Sci. 2014;22(3):138-143.
Siqueira JF Jr, Rôças IN. Exploiting molecular methods to explore endodontic infections: Part 1—current molecular technologies for microbiological diagnosis. J Endod. 2005;31(6):411-423.
Lin LM, Pascon EA, Skribner J, et al. Clinical, radiographic, and histologic study of endodontic treatment failures. Oral Surg Oral Med Oral Pathol. 1991;71(5):603-611.
Ricucci D, Siqueira JF Jr. Biofilms and apical periodontitis: study of prevalence and association with clinical and histopathologic findings. J Endod. 2010;36(8):1277-1288.
Nagata JY, Soares AJ, Souza-Filho FJ, et al. Microbial analysis of pre- and postinstrumentation root canal samples using pyrosequencing in primary endodontic infections with periapical disease: a clinical study. J Endod. 2014;40(6):899-904.
Marsh PD. Dental plaque as a biofilm and a microbial community—implications for health and disease. BMC Oral Health. 2006;6(Suppl 1):S14.
Khalifa W, Nokkaew N, Yang J, et al. Comparison of bacterial viability after exposure to endodontic irrigants. J Endod. 2019;45(7):858-862.
Kumar PS. Molecular and microbiologic insights into the oral microbiome and periodontal diseases. Periodontol 2000. 2013;64(1):226-237.
Sedgley CM, Lennan SL, Clewell DB. Prevalence, phenotype, and genotype of oral enterococci. Oral Microbiol Immunol. 2004;19(2):95-101.
Fouad AF, Barry J, Russo J, et al. PCR-based identification of bacteria associated with endodontic infections. J Clin Microbiol. 2002;40(9):3223-3231.
Neelakantan P, Romero M, Vera J, et al. Biofilms in endodontics: current status and future directions. Int J Mol Sci. 2017;18(8):1748.
Wang QQ, Zhang CF, Chu CH, et al. Prevalence of enterococcus faecalis in saliva and filled root canals of teeth associated with apical periodontitis. Int J Oral Sci. 2012;4(1):19-23.
Zehnder M. Root canal irrigants. J Endod. 2006;32(5):389-398.
Ercan E, Dalli M, Dülgergil CT. In vitro assessment of the antibacterial efficiency of various endodontic irrigation solutions. J Appl Oral Sci. 2006;14(5):341-345.
Siqueira JF Jr, Rôças IN. Clinical implications and microbiology of bacterial persistence after treatment procedures. J Endod. 2008;34(11):1291-1301.
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