Evaluation of the influence of influence of endodontic treatment in patients with periodontal chronic disease through microbiological analysis by PCR quantification of endotoxin and proinflammatory cytokines

Author(s)

Thais Mageste Duque

• Document type: Master's Dissertation
• Press: Piracicaba, SP.
• Institution: Universidade Estadual de Campinas (UNICAMP). Faculdade de Odontologia de Piracicaba
• Defense date: 2013-02-22

Examining board members

Brenda Paula Figueiredo de Almeida Gomes; Celso Neiva Campos; Alexandre Augusto Záia

• Advisor: Brenda Paula Figueiredo de Almeida Gomes

Abstract

The chronic periodontal disease affects the supporting structures of the teeth and is a major cause of tooth loss in adults. This long-term illness that does not regress with therapy, may have already caused an irreversible change in the pulp, even when clinically responds to pulp sensitivity tests. The lipopolysaccharide (LPS) present in the walls of gram negative bacteria and inflammatory cytokines act as factors for stimulating the immune response, causing bone destruction and increasing inflammation. The objective of this study was to evaluate the influence of endodontic treatment in patients with chronic periodontitis using microbiological analysis by pcr, endotoxin quantification and monitoring of inflammatory cytokines in periodontal pockets and root canals. Samples were taken from 15 teeth with chronic periodontal involvement and positive pulp sensitivity was selected. After chemical mechanical preparation (MCP), the teeth were divided into three groups: G1- one visit (n = 5), GII - Ca(OH)2 associated with chlorhexidine gel 2% (CHX-gel) (n = 5) ; GIII - Ca(OH)2 + saline solution (SS). Samples were collected at three different times in endodontic therapy: initial, after MCP and after intracanal medication (ICM) for 30 days, through paper cones apirogênics / sterile. The microorganisms were identified by PCR simple molecular technique (16S rDNA) by using specific primers for Fusobacterium nucleatum, Prevotella intermédia, red complex (Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia), and Parvimonas micras. For quantification of endotoxin was performed the Limulus Amebocyte Lysate (LAL) test and for the quantification of cytokines IL-1 ?, IL-1 ?, TNF ? and PGE2 was used ELISA immunoenzymatic assay. The results showed that after 1 year, 15 of the treated teeth, 5 were extracted. The other 10 teeth showed improvement related to probing depth and mobility. In the initial periodontal pockets, at least one of the microorganisms of the red complex were present in 12/15 samples and 3 together were present in 6/15. Canals samples showed the presence of Parvimonas micra and Fusobaterium nucleatum detected in 80% of cases. There was no association between the bacteria found in the pockets and root canals. After the CMP and use of ICM for 30 days, there was reduction of microbial content in both places. The initial endotoxin concentration in periodontal pockets were high (192.81 EU / ml), but after the CMP the reduction (19.65 EU / ml) was statistically significantly observed. Root canals showed low concentrations of endotoxin (0.1 EU / mL), being compatible with state of the pulp and maintained for other samples. Accordin the tested medications, reduction of LPS in the pockets showed better results with the use of Ca(OH)2 associated to saline solution (p <0.05). There was a reduction of all inflammatory cytokines when compared to the initial samples and after use of the dressing in pocktes and canals, being statistically significant in relation to IL1? and TNFa. There was no positive correlation between endotoxin and cytokines in periodontal pockets. In conclusion the red complex is present in the periodontal disease and that the presence of dressing decreases the concentration of inflammatory cytokines and LPS in root canals and periodontal pockets (AU)

• FAPESP's process: 10/13984-0 - Microbiological analysis for PCR, quantification of endotoxins and the progress of inflamatory process in patients with Cronical Periodontal Disease
• Grantee: Thais Mageste Duque
• Support Opportunities: Scholarships in Brazil - Master