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| Author(s): |
Marlos Barbosa Ribeiro
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | Piracicaba, SP. |
| Institution: | Universidade Estadual de Campinas (UNICAMP). Faculdade de Odontologia de Piracicaba |
| Defense date: | 2015-06-02 |
| Examining board members: |
Brenda Paula Figueiredo de Almeida Gomes;
Fabíola Bastos de Carvalho;
Alexandre Augusto Záia
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| Advisor: | Brenda Paula Figueiredo de Almeida Gomes; Caio Cezar Randi Ferraz |
| Abstract | |
The knowledge of the microbial community and its antigenic potential is important for understanding the pathogenesis of apical periodontitis in endodontically treated teeth. The objectives of this study were: 1) Characterize the microbiota associated to endodontic failure through the biochemical test, Nested PCR and partial sequencing of gene 16S rRNA.; 2) Quantify endotoxin (LPS) by Limulus Amebocyte Lysate test and pro-inflammatory cytokines IL-1?, IL-1?, TNF-? and PGE2 by enzyme-linked immunosorbent assay, correlating them with the patients' clinical and radiographic features; 3) To evaluate the effect of chemo-mechanical preparation (CMP) and intracanal medications (ICM) in reducing microorganisms, LPS and pro-inflammatory cytokines. Samples of endodontic content of 20 single-rooted teeth were collected before (C1), after CMP (C2), after the use of 17% EDTA (C3) agitated with ultrasound, and after ICM (C4). Sterile/apyrogenic materials and a aseptic techniques were used during the samples' collection. After processing the samples, the data were tabulated and analyzed by statistical testes Chi-square, Spearman correlation, and analysis of variance (Tukey-Kramer). The results indicated the presence of a mixed microbiota presenting Gram positive and Gram negative bacteria. Four different phyla were identified: Actinobacteria, Firmicutes, Proteobacteria and Fusobacterium. Enterococcus faecalis was the most prevalent species, identified phenotypically (19/158) and genotypically (42/158). Sequencing was able to identify 10.12% of the samples at the genus level and 89.88% at the species level, whereas the biochemical test identified 13.29% and 51.26%, respectively. The most prevalent bacteria by Nested PCR were: Enterococcus faecalis (97.5%), followed by Porphyromonas gingivalis (88.75%), Fusobacterium nucleatum (56.25%) and Aggregatibacter actinomycetemcomitans (37.5%). CMP reduced bacteria in 96.69% (p<0.05), with a percentage reduction in 99.3% for 2% CLX gel and in 92.1% for 6% NaOCl (p>0.05). EDTA 17% showed only a further effect on bacterial reduction in the 6% NaOCl group (p<0.05). Only 2% CLX gel was effective in reducing endotoxin compared to 6% NaOCl and 17% EDTA (p<0.05). CMP regardless of chemical substance used, was effective in reducing the levels of pro-inflammatory cytokines IL-1? (88.14%), IL-1? (89.33%), TNF-? (89.85%) and PGE2 (48.60%) (p<0.05).The intracanal medication not show additive effect in reducing bacteria and proinflammatory cytokines in both groups (p<0.05); and only presented action against LPS in the Ca(OH)2 + 2% CLX gel group (p<0.05). There was a positive correlation between bacteria, LPS, pro-inflammatory cytokines and clinical and radiographic features (p<0.05). In conclusion, CMP presented a significant effect in reducing the infectious content of root canal of teeth with endodontic failure, while ICM showed no significant additive effect in reducing these levels (AU) | |
| FAPESP's process: | 12/23697-4 - Microbial diversity and quantification of endoxinas and proinflammatory cytokines in secondary/persistent infections with periapical lesions |
| Grantee: | Marlos Barbosa Ribeiro |
| Support Opportunities: | Scholarships in Brazil - Master |