Title

Culture-Independent Characterization of the Microbiome of Healthy Pulp

Authors

Christopher Widmer, Nova Southeastern University
Jorie L. Skutas, Nova Southeastern UniversityFollow
Cole Easson, Nova Southeastern UniversityFollow
Jose Lopez, Nova Southeastern UniversityFollow
Calvin Torneck, Nova Southeastern University
Michael Flax, Nova Southeastern University
Taner Cem Sayin, Nova Southeastern University

Document Type

Article

Publication Date

5-2018

Publication Title

Journal of Endodontics

Keywords

Endodontic commensalism, Endodontic mutualism, Illumina, Pulp microbiome, Pulpal symbiont

ISSN

0099-2399

First Page

1

Last Page

8

Abstract

Introduction: Advances in culture-independent molecular biotechnologies have driven a greater appreciation for the function of mutualistic microorganisms in the maintenance of states of health in humans. The purpose of this study was to test the long-held hypothesis that healthy pulp lack bacteria.

Methods: Strict inclusion criteria were used to identify 10 pristine teeth from 10 healthy patients that were scheduled to be electively extracted in compliance with an orthodontic treatment plan. Using a rigorous disinfection protocol to isolate the operating field, the pulp space was accessed, and pulp tissue was collected in vivo from each tooth using a barbed broach. Genomic DNA was extracted from each pulp sample and analyzed for the presence of bacterial DNA using universal 16S ribosomal RNA polymerase chain reaction primers and MiSeq sequencing (Illumina, San Diego, CA) of community amplicons.

Results: One hundred percent (10/10) of the tested pulp tissues demonstrated the presence of bacterial DNA, with a mean of 343 operational taxonomic units per sample (range, 191–479). These were derived from 12 genera in which Ralstonia, Actinetobacter, and Staphylococcus were predominant (43%–78% of total community). None of the negative-field controls and none of the instruments used in the study tested positive for the presence of contaminating DNA.

Conclusions: This study presents evidence to support the conclusion that the pulp spaces of pristine healthy teeth contain detectable bacterial DNA.

Comments

©2018 American Association of Endodontists

NSUWorks Citation

Widmer, Christopher; Jorie L. Skutas; Cole Easson; Jose Lopez; Calvin Torneck; Michael Flax; and Taner Cem Sayin. 2018. "Culture-Independent Characterization of the Microbiome of Healthy Pulp." Journal of Endodontics , (): 1-8. doi:10.1016/j.joen.2018.03.009.

ORCID ID

0000-0002-1637-4125

ResearcherID

F-8809-2011

DOI

10.1016/j.joen.2018.03.009