Clinical, microbiological, and immunological effects of systemic probiotics in periodontal treatment: study protocol for a randomized controlled trial

Belen Retamal-Valdes, Wim Teughels, Laryssa Macedo Oliveira, Rebeca Nascimento da Silva, Aretuza Fritoli, Patricia Gomes, Geisla Mary Silva Soares, Natalie Temporão, Camila Pinheiro Furquim, Poliana Mendes Duarte, Helio Doyle, Marcelo Faveri, Luciene Cristina Figueiredo, Magda Feres, Belen Retamal-Valdes, Wim Teughels, Laryssa Macedo Oliveira, Rebeca Nascimento da Silva, Aretuza Fritoli, Patricia Gomes, Geisla Mary Silva Soares, Natalie Temporão, Camila Pinheiro Furquim, Poliana Mendes Duarte, Helio Doyle, Marcelo Faveri, Luciene Cristina Figueiredo, Magda Feres

Abstract

Background: The association of scaling and root planing (SRP) with systemic metronidazole (MTZ) plus amoxicillin (AMX) has shown to be an effective treatment protocol, particularly for periodontitis stages III and IV, generalized. More recently, probiotics have also been suggested as a promising adjunctive treatment for periodontal diseases due to their antimicrobial and anti-inflammatory properties. Therefore, the aim of this randomized clinical trial (RCT) is to evaluate the clinical, microbiological, and immunological effects of probiotics as adjuncts to SRP alone or with MTZ+AMX in the treatment of periodontitis.

Methods: Subjects with periodontitis are being randomly assigned to receive (i) SRP alone, or with (ii) two probiotic lozenges/day for 90 days (Prob), (iii) MTZ (400 mg) and AMX (500 mg) thrice a day (TID) for 14 days (MTZ+AMX), or (iv) Prob and MTZ+AMX. Subjects are being monitored for up to 12 months post-treatment. Nine subgingival plaque samples per patient are being collected at baseline and at 3, 6, and 12 months post-therapy and analyzed by checkerboard DNA-DNA hybridization for 40 bacterial species. Peripheral blood and gingival crevicular fluid (GCF) of four randomly selected periodontal sites will be analyzed by means of a multiplex fluorescent bead-based immunoassay for 17 cyto/chemokines.

Statistical analyses: The significance of differences in each group (over the course of the study) will be sought using repeated measures ANOVA or Friedman tests and among groups (at each time point) using either ANOVA/ANCOVA or Kruskal-Wallis tests, depending on normality of the data. The chi-square test will be used to compare differences in the frequency of subjects achieving the clinical endpoint for treatment (≤ 4 sites with PD ≥ 5 mm) at 1 year and of self-perceived adverse effects. A stepwise forward logistic regression analysis will be performed in order to investigate the impact of different predictor variables on the percentage of patients achieving the clinical endpoint for treatment. The Number Needed to Treat (NNT) with different treatment protocols will be also calculated. Statistical significance will be set at 5%.

Trial registration: ClinicalTrials.gov NCT03733379. Registered on November 7, 2018.

Keywords: Amoxicillin; Metronidazole; Periodontitis; Probiotic; Scaling and root planing; Treatment.

Conflict of interest statement

This study is being supported by BioGaia AB Company, Stockholm, Sweden. The authors certify that have no commercial or associative interest that represents a conflict of interest in connection with the manuscript.

Figures

Fig. 1
Fig. 1
Experimental design of the study. AMX, amoxicillin (500 mg, 3 times × day); CLIN, clinical assessment; OHI, oral hygiene instruction; IMUNO, immunological assessment; MICRO, microbiological assessment; MTZ, metronidazole (400 mg, 3 times × day); Prob, probiotic; PMT, periodontal maintenance therapy; SRP, scaling and root planing
Fig. 2
Fig. 2
Schedule of volunteers’ enrolment, interventions, and assessments according to the SPIRIT statement. AMX, amoxicillin; Mo, months; MTZ, metronidazole

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