Characterization of Root Canal Microbiota in Teeth Diagnosed with Irreversible Pulpitis

Abstract

Introduction: Previous studies have shown that in teeth presenting with symptoms of irreversible pulpitis (IP), bacteria and their by-products driving inflammation are confined mainly within the coronal pulpal tissue. The present study aimed to determine the presence and identity of bacteria within pulps presenting with clinical symptoms of IP using molecular methods.

Methods: Samples were obtained from 30 adult patients presenting to the dental emergency department with signs and symptoms of IP. After meticulous surface decontamination, the pulp space was accessed, and clinical samples were collected from inflamed pulp tissue using sterile paper points. Genomic DNA was extracted from the clinical samples, and quantification of bacteria was performed using quantitative polymerase chain reaction targeting the conserved 16S ribosomal RNA (rRNA) gene. To characterize the microbial composition, the V3-V5 hypervariable regions of the 16S rRNA gene were amplified and subjected to next-generation sequencing on the MiSeq platform (Illumina, San Diego, CA).

Results: Of the 30 teeth that presented with IP, half of the intracanal samples had a substantial bacterial load (16S rRNA copies) within the IP vital pulp as determined by quantitative polymerase chain reaction. Next-generation sequencing microbial identification was successful in 7 intracanal samples and yielded 187 bacterial operational taxonomic units within the IP samples. The most abundant genera observed among the vital cases were Veillonella (16%), Streptococcus (13%), Corynebacterium (10%), Cutibacterium (9.3%), and Porphyromonas (5.7%).

Conclusions: The current study highlighted the evidence of vital teeth diagnosed as IP harboring considerable bacterial loads and composed of genera reflective of established endodontic pathology and thus may offer insights into the initial events preceding pulpal necrosis.

Keywords: Irreversible pulpitis; next-generation sequencing; quantitative polymerase chain reaction.