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Microbiological analysis and quantification of endotoxins and proinflammatory cytokines from primarily infected root canals with apical periodontitis


The main virulence factor of gram-negative bacteria is represented by the release of endotoxin (LPS) presented in the outer layers of their cell walls. The accumulation of these bacterial components in the root canal and periradicular tissues should stimulate macrophages present in the area to produce cytokines such as IL-1 and TNF-± which are potent stimulators of bone resorption. The aim of this study is to isolate and identify gram-negative anaerobic species through biochemical methods and PCR from root canals of teeth with necrotic pulp and apical periodontitis, evaluating the effect of chemomechanical preparation (CMP) with 2% chlorhexidine gel and 17% EDTA, using or not ultrasound. The reduction of microorganisms and LPS, correlating the microbiota and endotoxin content with the expression of pro-inflammatory cytokines IL-1a, IL-1b, TNF± and PGE-2. Twenty-four root canals were selected with pulp necrosis and presence of periapical lesion. Endotoxins and microbiological samples will collect Samples were collected before and after CMP, before and after using 17% EDTA, with or without application of ultrasound, and finally after seven days of the first session. Samples for cytokines quantification will also collect by passing a paper point 1 mm beyond the apex, after the PQM and after 7 days, The microbiological samples will be diluted, plated and incubated for further isolation and identification. PCR will be performed using specific primers. A limulus amebocyte lysate (LAL) assay will be used to quantify endotoxins Cytokines collected from apex will be quantified using specific kits. The values obtained will be tabulated and statistically analyzed. (AU)

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Scientific publications (13)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
MARINHO, A. C. S.; MARTINHO, F. C.; GONCALVES, L. M.; RABANG, H. R. C.; GOMES, B. P. F. A.. Does the Reciproc file remove root canal bacteria and endotoxins as effectively as multifile rotary systems?. International Endodontic Journal, v. 48, n. 6, p. 542-548, . (13/02402-9, 10/13498-9, 10/17877-4, 13/17928-6, 10/19136-1, 11/09047-4)
MARTINHO, FREDERIC C.; LEITE, FABIO R. M.; CHIESA, WANDERSON M. M.; NASCIMENTO, GUSTAVO G.; FERES, MAGDA; GOMES, BRENDA P. F. A.. Signaling Pathways Activation by Primary Endodontic Infectious Contents and Production of Inflammatory Mediators. JOURNAL OF ENDODONTICS, v. 40, n. 4, p. 484-489, . (10/19136-1, 13/02402-9, 10/17877-4)
FREDERICO C. MARTINHO; FÁBIO R. M. LEITE; LETÍCIA M.M. NÓBREGA; MARCOS S. ENDO; GUSTAVO G. NASCIMENTO; RICHARD P. DARVEAU; BRENDA P. F. A. GOMES. Comparison of Fusobacterium nucleatum and Porphyromonas gingivalis Lipopolysaccharides Clinically Isolated from Root Canal Infection in the Induction of Pro-Inflammatory Cytokines Secretion. Brazilian Dental Journal, v. 27, n. 2, p. 202-207, . (11/09047-4, 10/17877-4, 10/19136-1)
MARINHO, ARIANE C. S.; MARTINHO, FREDERIC C.; LEITE, FABIO R. M.; NASCIMENTO, GUSTAVO G.; GOMES, BRENDA P. F. A.. Proinflammatory Activity of Primarily Infected Endodontic Content against Macrophages after Different Phases of the Root Canal Therapy. JOURNAL OF ENDODONTICS, v. 41, n. 6, p. 817-823, . (10/19136-1, 13/02402-9, 10/17877-4)
MARINHO, ARIANE C. S.; POLAY, ANA R. O.; GOMES, BRENDA P. F. A.. Accuracy of Turbidimetric Limulus Amebocyte Lysate Assay for the Recovery of Endotoxin Interacted with Commonly Used Antimicrobial Agents of Endodontic Therapy. JOURNAL OF ENDODONTICS, v. 41, n. 10, p. 1653-1659, . (10/19136-1, 13/02402-9, 10/17877-4)
HERRERA, D. R.; SILVA, E. J. N. L.; SANTOS, C. C.; ZAIA, A. A.; FERRAZ, C. C. R.; ALMEIDA, J. F. A.; GOMES, B. P. F. A.. Root canal content from primary endodontic infection and upregulation of gelatinases in fibroblast cells. International Endodontic Journal, v. 48, n. 12, p. 1168-1174, . (10/19136-1, 10/17877-4, 10/13498-9)
SALUSTIANO MARINHO, ARIANE CASSIA; MARTINHO, FREDERICO CANATO; ZAIA, ALEXANDRE AUGUSTO; RANDI FERRAZ, CAIO CEZAR; FIGUEIREDO DE ALMEIDA GOMES, BRENDA PAULA. Monitoring the effectiveness of root canal procedures on endotoxin levels found in teeth with chronic apical periodontitis. Journal of Applied Oral Science, v. 22, n. 6, p. 490-495, . (13/02402-9, 10/17877-4, 10/19136-1, 11/09047-4, 10/13743-3)
ARIANE CASSIA SALUSTIANO MARINHO; FREDERICO CANATO MARTINHO; ALEXANDRE AUGUSTO ZAIA; CAIO CEZAR RANDI FERRAZ; BRENDA PAULA FIGUEIREDO DE ALMEIDA GOMES. Influence of the apical enlargement size on the endotoxin level reduction of dental root canals. Journal of Applied Oral Science, v. 20, n. 6, p. 661-666, . (10/19136-1, 10/17877-4)
ENDO, M. S.; MARTINHO, F. C.; ZAIA, A. A.; FERRAZ, C. C. R.; ALMEIDA, J. F. A.; GOMES, B. P. F. A.. Quantification of cultivable bacteria and endotoxin in post-treatment apical periodontitis before and after chemo-mechanical preparation. EUROPEAN JOURNAL OF CLINICAL MICROBIOLOGY & INFECTIOUS DISEASES, v. 31, n. 10, p. 2575-2583, . (10/17877-4, 10/19136-1)
HERRERA, D. R.; MARTINHO, F. C.; DE-JESUS-SOARES, A.; ZAIA, A. A.; FERRAZ, C. C. R.; ALMEIDA, J. F. A.; GOMES, B. P. F. A.. Clinical efficacy of EDTA ultrasonic activation in the reduction of endotoxins and cultivable bacteria. International Endodontic Journal, v. 50, n. 10, p. 933-940, . (10/17877-4, 13/23061-5)
SOUSA, EZILMARA L. R.; MARTINHO, FREDERICO C.; LEITE, FABIO R. M.; NASCIMENTO, GUSTAVO G.; GOMES, BRENDA P. F. A.. Macrophage Cell Activation with Acute Apical Abscess Contents Determined by Interleukin-1 Beta and Tumor Necrosis Factor Alpha Production. JOURNAL OF ENDODONTICS, v. 40, n. 11, p. 1752-1757, . (10/19136-1, 10/17877-4, 11/09047-4)
GOMES, BRENDA P. F. A.; ENDO, MARCOS S.; MARTINHO, FREDERICO C.. Comparison of Endotoxin Levels Found in Primary and Secondary Endodontic Infections. JOURNAL OF ENDODONTICS, v. 38, n. 8, p. 1082-1086, . (10/17877-4, 10/19136-1)
MARTINHO, FREDERICO C.; CHIESA, WANDERSON M. M.; LEITE, FABIO R. M.; CIRELLI, JONI A.; GOMES, BRENDA P. F. A.. Correlation between Clinical/Radiographic Features and Inflammatory Cytokine Networks Produced by Macrophages Stimulated with Endodontic Content. JOURNAL OF ENDODONTICS, v. 38, n. 6, p. 740-745, . (10/17877-4, 10/19136-1)

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