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| Author(s): |
Lisa Danielly Curcino Araujo
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | Ribeirão Preto. |
| Institution: | Universidade de São Paulo (USP). Faculdade de Odontologia de Ribeirão Preto (PCARP/BC) |
| Defense date: | 2020-12-11 |
| Examining board members: |
Léa Assed Bezerra da Silva;
Paulo Sérgio Cerri;
Flavia Aparecida Chaves Furlaneto Messora;
Paulo Nelson Filho
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| Advisor: | Léa Assed Bezerra da Silva |
| Abstract | |
Probiotics are living microorganisms that, when administered in adequate amounts, confer health benefits to the host. Research regarding the use of probiotics to treat diseases in the oral cavity has shown relevant results. However, few studies report the effect of using probiotics on pulp infections and neither in vivo study has used probiotics as an irrigation solution for root canals. Thus, the objective of the present study was to evaluate, in vitro, the antimicrobial activity of the probiotic Bifidobacterium animalis subsp. lactis HN019, using the well technique, against 10 microorganisms that can be found involved in endodontic infections. In addition, it was evaluated in vivo, through microscopic analysis, the effect of this probiotic when used as an irrigation solution for root canals of rat\'s teeth with induced periapical lesions. For in vitro analysis, probiotic antimicrobial activity was performed on Streptococcus mutans, Streptococcus sobrinus, Lacticaseibacillus caseii, Enterococcus faecalis, Staphylococcus aureus, Candida albicans, Porphyromonas gingivalis, Porphyromonas endodontalis, Prevotella intermedia e Fusobaterium nucleatum. After 48 to 72 hours of incubation of the petri dish containing culture medium, the strains of the microorganisms and the probiotic, the dishes were examined to assess the uniformity of microbial growth, the presence of contaminants and the halo of inhibition. After visual inspection, the inhibition halo was read with the aid of a digital caliper using a reflected light source to illuminate the inverted plate on a black and opaque background after removing the cover. Thus, 3 values of each bacterial inoculum were obtained, which were added and divided by three to obtain the average of the values. For the in vivo evaluation, the animals were divided into six groups according to the irrigating solution used in the root canals after coronary opening and exposure of the pulp tissue, as follows: Group I (4 animals / 8 teeth) healthy teeth; Group II (4 animals / 4 teeth): 2.5% sodium hypochlorite as an irrigating solution (3 days); Group III (4 animals / 4 teeth): Probiotic as an irrigating solution (3 days); Group IV (12 animals / 24 teeth): Periapical lesion without any treatment (7, 21 and 42 days); Group V (12 animals / 24 teeth): 2.5% sodium hypochlorite as an irrigating solution (7, 21, 42 days); Group VI (12 animals / 24 teeth): Probiotic as an irrigating solution (7, 21, 42 days). The probiotic Bifidobacterium animalis subsp. lactis (B. lacti) HN019 was used as an irrigating solution, formulated by adding the probiotic at 2.7x 109 CFU in aqueous medium, plus 2.0% of carboxymethylcellulose, with a final volume of 100mµL. For local administration, the root canals were irrigated on day 0 and days 3, 7, 14, 21, 28 and 35 days after induction of the periapical lesion. After euthanizing the animals, the mandibles were submitted to histotechnical processing, the slides stained in hematoxylin and eosin (HE) and analyzed in optical microscopy to describe the characteristics of the root canal content and the apical and periapical regions, in parallel to the morphometric analysis of the area periapical lesions, under fluorescent microscopy. Osteoclast counting was also performed, using histoenzymology for Tartrate Resistant Acid Phosphatase Activity (TRAP). The results obtained were submitted to statistical analysis through different tests according to the nature of the data. For all analyzes, the significance level of 5% was adopted. The results of the in vitro study demonstrated that the probiotic B. animalis subsp. lactis HN019 promoted the inhibition of all strains of the pathogens evaluated, with the exception of Candida albicans, demonstrating antimicrobial activity on these microorganisms. The results of the in vivo study of the analysis of the histopathological parameters showed that groups III and VI, with probiotic as an irrigating solution, presented better results when compared to the group without treatment and with the irrigation of sodium hypochlorite, showing less inflammatory cells and smaller lesion area. Regarding the number of osteoclasts, a decrease was observed only in group III, at 3 days (p=0.0425), and in group VI at 21 days (p=0.0043). After the methodologies and the results obtained, it can be concluded that the probiotic Bifidobacterium animalis subsp. lactis HN019 showed in vitro antimicrobial activity against microorganisms present in endodontic infections, besides presenting, microscopically, a therapeutic potential for local administration as an irrigating solution in the progression of periapical lesion, when compared to sodium hypochlorite. (AU) | |
| FAPESP's process: | 18/22038-3 - Microscopic analysis of the genesis and progression of periapical lesions after systemic and local use of probiotic in mice |
| Grantee: | Lisa Danielly Curcino Araujo |
| Support Opportunities: | Scholarships in Brazil - Master |