Machine Learning Study in Caries Markers in Oral Microbiota from Monozygotic Twin Children.

In recent years, the etiology of caries has evolved from a simplistic infectious perspective based on Streptococcus mutans and/or Lactobacillus activity, to a multifactorial disease involving a complex oral microbiota, the human genetic background and the environment. The aim of this work was to identify bacterial markers associated with early caries using massive 16S rDNA. To minimize the other factors, the composition of the oral microbiota of twins in which only one of them had caries was compared with their healthy sibling. Twenty-one monozygotic twin pairs without a previous diagnosis of caries were recruited in the context of their orthodontic treatment and divided into two categories: (1) caries group in which only one of the twins had caries; and (2) control group in which neither of the twins had caries. Each participant contributed a single oral lavage sample in which the bacterial composition was determined by 16S rDNA amplification and further high-throughput sequencing. Data analysis included statistical comparison of alpha and beta diversity, as well as differential taxa abundance between groups. Our results show that twins of the control group have a closer bacterial composition than those from the caries group. However, statistical differences were not detected and we were unable to find any particular bacterial marker by 16S rDNA high-throughput sequencing that could be useful for prevention strategies. Although these results should be validated in a larger population, including children from other places or ethnicities, we conclude that the occurrence of caries is not related to the increase of any particular bacterial population.

Photodynamic therapy in endodontic root canal treatment significantly increases bacterial clearance, preventing apical periodontitis.

OBJECTIVE: To analyze the antimicrobial activity of photodynamic therapy as an adjunct to conventional endodontic treatment, particularly against Enterococcus faecalis. METHOD AND MATERIALS: A total of 42 single-rooted teeth obtained from 33 patients with apical periodontitis were included. Sampling was developed in three stages: (1) immediately after accessing the root canal, (2) after chemical and mechanical instrumentation, and finally, (3) after photodynamic therapy application. The bacterial load of each sample was quantified by seeding on blood agar plates and selective M-Enterococcus agar. All growing colonies were identified using MALDI-TOF (Bruker; matrix-assisted laser desorption/ionization time-of-flight), and the entire bacterial microbiota composition was determined in the first sample by PCR-DGGE (polymerase chain reaction denaturing gradient gel electrophoresis), using 16 rDNA primers and selective nucleotide sequencing. RESULTS: The endodontic therapy obtained a mean reduction in the cultivable bacterial load of 1.12 log, whereas the photodynamic therapy combination significantly increased the bacterial clearance (P < .0001). Viable cells of E faecalis were detected in 16.6% of root canals, with a mean value of 93 CFU per tooth, which was reduced to 67 and 9 CFU/tooth after conventional endodontic and photodynamic therapy treatments, respectively. Molecular E faecalis detection demonstrated that this species was present in 23.2% of baseline samples. DGGE analysis demonstrated the existence of a more complex microbiota than those observed using classical cultures. CONCLUSION: Photodynamic therapy as an adjunct to root canal treatment produces a significant reduction in E faecalis bacterial load, and it should be considered in the prevention of apical periodontitis.

Forensic identification in teeth with caries.

Human teeth are biological structures that resist extreme conditions thus becoming a useful source of DNA for human forensic identification purposes. When it is possible, forensic prefer only non-damaged teeth whereas those with cavities are usually rejected to avoid both external and internal bacterial contamination. Cavities are one of the most prevalent dental pathology and its incidence increases with ageing. The aim of this study was to validate the use of teeth with cavities for forensic identification. A total of 120 individual teeth from unrelated patients (60 healthy and 60 with cavities, respectively) extracted by a dentist as part of the normal process of treatment, were submitted for further analysis. Dental pulp was obtained after tooth fragmentation, complete DNA was extracted and the corresponding human identification profile was obtained by the AmpFlSTR® NGM SElect™ kit. Cariogenic microbiota was determined by PCR-DGGE with bacterial universal primers and bands were excised, re-amplified and sequenced. From the 120 dental pieces analyzed, a defined genetic profile was obtained in 81 (67.5%) of them, with no statistical differences between the healthy and the cavities-affected teeth. Statistical association between teeth status, DNA content and genetic profiles was not observed. Complex bacterial communities were only detected in the cavities group, being the Streptococcus/Enterococcus, and Lactobacillus genera the most represented. We conclude that teeth with cavities are as valid as healthy dental pieces for forensic human identification. Moreover, the severity of the cariogenic lesion as well as associated bacterial communities seems not to influence the establishment of human dental profiles.