Characterization of Lactobacilli isolated from carious dentin after selective caries removal and cavity sealing.

Nailê Damé-Teixeira, Laís Daniela Ev, Luciana Bitello-Firmino, Vanessa Kern Soares, Raquel Soares Dalalba, Ariel Goulart Rup, Marisa Maltz, Clarissa Cavalcanti Fatturi Parolo

Journal: Archives of oral biology 2021;121():104988

PMID: 33242691

Full Text on Elsevier Science View on PubMed

Abstract

OBJECTIVE

To characterize lactobacilli isolated from residual carious dentin after selective caries removal (SCR), by observing the changes detected in their prevalence, diversity, and cariogenic potential after starvation stress caused by cavity sealing (CS).

DESIGN

Lactobacilli were cultured from carious dentin lesions (n = 16 patients) treated in a clinical trial, three months before and after CS. Presumptive lactobacilli were selected, isolated, and analyzed by Gram staining. Housekeeping gene sequences were used to identify the species (groEL, rpoA, pheS, and 16S rRNA).

RESULTS

N = 86 Lactobacillus spp. (n = 41 before and n = 45 after sealing) were genotyped by AP-PCR and analyzed for their cariogenic potential (acid production and acid tolerance). The proportion of lactobacilli to the total anaerobic counts was high, and a significant decrease was observed after sealing (median before sealing = 78.9; 25th-75th = 60.25-97.35; median after sealing = 0.00; 25th-75th = 0.00-77.08; p = 0.001). L. paracasei was the most prevalent species of lactobacilli in carious dentin (p = 0.02). L. rhamnosus prevalence increased to a proportion similar to L. paracasei after CS (p = 0.001). A total of 28 and 14 different genotypes were found before and after CS, respectively. There was no difference between the L. paracasei and the L. rhamnosus isolated from carious dentin, neither regarding acid production nor acid tolerance.

CONCLUSIONS

Although there was a significant reduction in lactobacilli in the residual carious dentin after SCR, some strains were capable of surviving after three months of CS. However, the sealed available nutrients are low and not sufficient for caries progression. Also, we believe that a longer follow up period may eliminate all the residual lactobacilli. L. paracasei prevailed in carious dentin in a proportion similar to L. rhamnosus in the sealed dentin. Characterization of lactobacilli after SCR and sealing may help the understanding the importance of genotyping of lactobacilli in carious microbiota.

Address: Department of Dentistry, School of Health Sciences, University of Brasilia, Federal District, Brazil. Electronic address: [email protected].; Department of Social and Preventive Dentistry, School of Dentistry, Federal University of Rio Grande do Sul, Porto Alegre, Brazil.

DOI: 10.1016/j.archoralbio.2020.104988