Supra and subgingival microbiota diversity analysis and protein profile of saliva and enamel film of individuals with chronic periodontitis and microbial succession

• Grant number: 16/13159-6
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: August 01, 2016
• End date: July 31, 2021
• Field of knowledge: Health Sciences - Dentistry - Periodontology
• Principal Investigator: Marcia Pinto Alves Mayer
• Grantee: Dione Kawamoto
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 15/18273-9 - New strategies for the control of periodontitis, AP.TEM

Abstract

Chronic periodontitis (CP) is associated with oral microbiota dysbiosis, where the microbial succession determined mainly by P. gingivalis leads to the formation of pathogenic biofilms, able to induce a destructive and inflammatory response in the host. Saliva plays an important role in maintaining homeostasis and oral saliva biomolecules can provide information about various organs and systems, whereas its composition influences the microbial composition and reflects the immune response. Hypothesis: patients with CP have differences in salivary composition in relation to individuals with healthy periodontium (HP) resulting in differences in the acquired enamel pellicle (AEP) that reflects in the oral microbiota. These differences in the saliva composition may interfere with other processes such as immunomodulatory capacity and antimicrobial activity. Patients with CP and controls will be evaluated regarding diversity of the oral biofilm microbiota and faeces, saliva and acquired pellicle protein profiles, and salivary cytokines. In addition, the influence of AEP in interfering with adhesion and biofilm formation will be determined. The immunomodulatory effect and antimicrobial activity of saliva, will be determined in both groups. Methods: individuals with CP and control subjects (HP) will be selected. Supra and subgingival biofilm, stimulated /unstimulated and parotid saliva, EAP, and feces will be sampled. The diversity of microbial biofilms and faeces will be performed after amplification of 16S rRNA followed by high-performance sequencing. The proteins profiles will be determined by mass spectrometry. The saliva cytokines will be determined using Magpix system. Saliva immunemodulatory activity of PC and HP patients will be determined in macrophages activated with LPS, and the antimicrobial activity determined by viable qPCR . The role of the EAP composition influencing adhesion / biofilm and of saliva to aggregate microorganisms will be determined microcosm biofilm assay with EAP formed with saliva, followed by assays with isolated bacteria. The results of the different techniques will be jointly evaluated in order to select potential biomarkers represented by salivary proteins, genetic variants and microbial groups in the oral cavity and faeces presented differentially between PC and HP. We hope to show that differences in the saliva composition are related to differences in oral and fecal microbiome, with reflect in the periodontal tissues conditions. (AU)

Scientific publications

(6)

(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)

AMADO, PAMELA PONTES PENAS; KAWAMOTO, DIONE; ALBUQUERQUE-SOUZA, EMMANUEL; FRANCO, DIEGO CASTILLO; SARAIVA, LUCIANA; CASARIN, RENATO CORREA VIANA; HORLIANA, ANNA CAROLINA RATTO TEMPESTINI; MAYER, MARCIA PINTO ALVES. Oral and Fecal Microbiome in Molar-Incisor Pattern Periodontitis. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, v. 10, OCT 8 2020. (15/00259-0, 16/14687-6, 16/13159-6, 15/18273-9)

SHIMABUKURO, NATALI; CATARUCI, AMALIA C. DE S.; ISHIKAWA, KARIN H.; OLIVEIRA, BRUNA E. DE; KAWAMOTO, DIONE; ANDO-SUGUIMOTO, ELLEN S.; ALBUQUERQUE-SOUZA, EMMANUEL; NICOLI, JACQUES R.; FERREIRA, CAROLINE M.; DE LIMA, JEAN; et al. Bifidobacterium Strains Present Distinct Effects on the Control of Alveolar Bone Loss in a Periodontitis Experimental Model. FRONTIERS IN PHARMACOLOGY, v. 12, SEP 24 2021. (17/22345-0, 15/18273-9, 16/13156-7, 18/25171-6, 16/14687-6, 17/16377-7, 16/13159-6)

KAWAMOTO, DIONE; LUCAS AMADO, PAMELA PONTES; ALBUQUERQUE-SOUZA, EMMANUEL; BUENO, MANUELA ROCHA; VALE, GLAUBER CAMPOS; SARAIVA, LUCIANA; ALVES MAYER, MARCIA PINTO. Chemokines and cytokines profile in whole saliva of patients with periodontitis. CYTOKINE, v. 135, p. 9-pg., 2020-11-01. (16/13159-6, 17/16377-7, 15/00259-0, 17/10829-3, 15/18273-9)

BUENO, MANUELA R.; ISHIKAWA, KARIN H.; ALMEIDA-SANTOS, GISLANE; ANDO-SUGUIMOTO, ELLEN S.; SHIMABUKURO, NATALI; KAWAMOTO, DIONE; MAYER, MARCIA P. A.. Lactobacilli Attenuate the Effect of Aggregatibacter actinomycetemcomitans Infection in Gingival Epithelial Cells. FRONTIERS IN MICROBIOLOGY, v. 13, p. 11-pg., 2022-05-04. (16/13159-6, 17/16377-7, 16/13156-7, 16/14687-6, 15/18273-9, 17/22345-0)

KAWAMOTO, DIONE; BORGES, RODRIGO; RIBEIRO, RODOLFO ALVARENGA; DE SOUZA, ROBSON FRANCISO; AMADO, PAMELA PONTES PENAS; SARAIVA, LUCIANA; HORLIANA, ANA CAROLINA RATTO TEMPESTINI; FAVERI, MARCELO; MAYER, MARCIA PINTO ALVES. Oral Dysbiosis in Severe Forms of Periodontitis Is Associated With Gut Dysbiosis and Correlated With Salivary Inflammatory Mediators: A Preliminary Study. FRONTIERS IN ORAL HEALTH, v. 2, p. 15-pg., 2021-10-11. (16/13159-6, 15/18273-9)

ISHIKAWA, KARIN HITOMI; MITA, DANIELA; KAWAMOTO, DIONE; NICOLI, JACQUES ROBERT; ALBUQUERQUE-SOUZA, EMMANUEL; LORENZETTI SIMIONATO, MARIA REGINA; ALVES MAYER, MARCIA PINTO. Probiotics alter biofilm formation and the transcription ofPorphyromonas gingivalisvirulence-associated genes. JOURNAL OF ORAL MICROBIOLOGY, v. 12, n. 1, JAN 1 2020. (16/13156-7, 15/18273-9, 16/14687-6, 16/13159-6)