Evaluation of Bacteriological Profile in the Apical Root Segment of the Patients with Primary Apical Periodontitis

Abstract

Introduction: Apical periodontitis usually results from bacterial accumulation and contamination occurring in the root-canal system, and extending beyond the apical foramen to involve the periapical tissues. Literature has a paucity of the studies that stress on the division and analysis of the pulp canal segments. The reason for this disparity might be the technique used for collecting the samples from the pulp canals. Hence, we carried out the present study to evaluate the microbial flora in the apical part of the roots with necrotic pulp canals. Materials and methods:The present study included the assessment of 40 freshly extracted teeth that had necrotized pulpal tissue along with the presence of periapical periodontal lesions. Removal of the soft tissue lesions attached to the root portion of the teeth along with apical periodontal lesions was done with the help of scalpel blade, after rinsing them with a sterile solution of saline. Thorough cleaning of the root surfaces was done with hydrogen peroxide followed by rapid disinfection with the help of sodium hypochlorite at varying concentrations. Sectioning of the root portion of all the specimens with the help of a disk was done perpendicular to the long axis of the teeth at a distance of roughly 5 to 6 mm from the teeth's apicalmost point. Cryotubes were used for transferring the specimens of apical portions containing 1 mL of buffer and were subjected to immediate frozen processing at a temperature of -20°C. A 10 K-type file was used for the initial collection of the samples followed by subsequent incubation of the files and paper pints in the incubation cabinet. Subsequent deoxyribonucleic acid (DNA) extraction from the samples was done following the procedure described by Siqueira et al. Paster et al's modification of the reverse-capture checkerboard assay was used in the present study. Semiquantitative data were used for overcoming the difficulties arising due to obtaining the counts of the polymerase chain reaction (PCR)-based analysis of specimens.Results: A positive result for the 16S ribosomal ribonucleic acid (rRNA) gene primer was observed only in two examined specimens of all the samples of the apical portion of the root canals in the present study. Negative result was shown by all the control group specimens, which were sterile samples. Presence of bacteria was confirmed by PCR in 38 out of 40 examined specimens. Amount of bacterial taxa, out of these 24 samples, ranged up to 6. Pseudoramibacter alactolyticus, Porphyromonas endodontalis, Dialister oral species, Bacteroidetes species, Streptococcus species, Olsenella uli, Synergistes species, Fusobacterium nucleatum, Parvimonas micra, Treponema denticola, and Filifactor alocis were the specific species detected. Bacteroidetes species was the only species that were detected at levels at or above 10 5 . Heavy bacterial infections were noticed in more than 45% of the cases at the periradicular part of the root canals. Conclusion:Microbial flora of the apical segment of the root with necroti...

Microbiota in the apical root canal system of tooth with apical periodontitis

“…Based on the 16S rRNA technique, complex bacterial populations were reported present in AP individuals, mostly classified within phyla of Proteobacteria , Firmicutes , Bacteriodites , Fusobacteia , and Actinobacteria [12, 16–23], where Proteobacteria , Firmicutes and Bacteriodites were always found prevalent. With 10 patients (10 extracted teeth) involved, Siqueira’s early study identified 187 phylotyes belonging to 84 genera and 10 phyla in apical root canal infections, where Proteobacteria and Firmicutes together occupied about two thirds of all sequences obtained [16].…”

Apical root canal microbiome associated with primary and posttreatment apical periodontitis: A systematic review

“…The final sample consisted of 21 studies (Figure 1). Ten studies investigated only primary infections (Baumgartner & Falkler, 1991; Dougherty et al., 1998; Ozok et al., 2012; Persoon et al., 2017; Rôças et al., 2010; Siqueira et al., 2009; Siqueira et al., 2011; Siqueira, Rôças, Alves, et al., 2004; Takahama et al., 2018; Tatikonda et al., 2017), 8 only posttreatment infections (Antunes et al., 2015; Pereira et al., 2017; Perez‐Carrasco et al., 2023; Ping et al., 2015; Siqueira et al., 2016; Siqueira et al., 2020; Subramanian & Mickel, 2009; Wang et al., 2012), and 3 included both types (Bouillaguet et al., 2018; Chugal et al., 2011; Qian et al., 2019). The search for grey literature using the Proquest database did not return any document related to the objective of the present study.…”

“…In one study, the root apex specimens were triturated using orthodontic pliers (Pereira et al., 2017). As for the other studies, 5 used endodontic files and paper points (Baumgartner & Falkler, 1991; Chugal et al., 2011; Siqueira et al., 2009; Siqueira, Rôças, Alves, et al., 2004; Tatikonda et al., 2017) and 2 studies used only endodontic files (Bouillaguet et al., 2018; Dougherty et al., 1998) to take samples from the root canal in the resected apical root segment; in the other 2 studies, samples were obtained by agitation and centrifugation of the root specimens (Subramanian & Mickel, 2009; Wang et al., 2012).…”

High-performing cross-dataset machine learning reveals robust microbiota alteration in secondary apical periodontitis

“…Previous investigations utilizing broad-range culture and 16S rRNA sequencing have identified a variety of relevant species into the microbial communities associated with necrotic root canals, primarily strict anaerobic bacteria from Peptostreptococcus, Prevotella, Porphyromonas, Fusobacterium, Eubacterium, and Actinomyces, along with facultative anaerobic Streptococci (Sakamoto et al, 2006;Tatikonda et al, 2017). Notably, the presence of these pathogens has been linked to primary apical periodontitis; whereas, secondary apical periodontitis exhibits distinct microbial populations, predominantly Gram-positive facultative anaerobes like Streptococcus, Lactobacillus, and Enterococcus (Siqueira and Roĉas, 2005;Siqueira et al, 2016;Qian et al, 2019).…”