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EFFECTS OF VIABLE, NON-VIABLE Lactobacillus reuteri AND SUPERNATANT ON PERIODONTAL DISEASE: AN in vitro AND in vivo STUDY

Grant number: 16/06946-1
Support type:Regular Research Grants
Duration: February 01, 2017 - January 31, 2019
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Ana Lia Anbinder
Grantee:Ana Lia Anbinder
Home Institution: Instituto de Ciência e Tecnologia (ICT). Universidade Estadual Paulista (UNESP). Campus de São José dos Campos. São José dos Campos , SP, Brazil
Assoc. researchers:Juliana Campos Junqueira ; Luciane Dias de Oliveira

Abstract

Periodontitis is multifactorial disease caused by biofilm action plus imune and adaptative host response, that results in periodontium destruction. Non surgical scaling and root planing, with or without antibiotics use, are the most used treatments. However, new therapies have been tested, some of them in experimental models, focusing in host response modulation. Probiotics are live microorganisms that confer a health benefit on the host, due to their antibiotic and anti-inflammatory effects. Our aim is to evaluate the antimicrobial effect of viable, non-viable and the cell culture supernatant of Lactobacillus reuteri Prodentis on Porphyromoas gingivalis, Fusobacterium nucleatum and Prevotella intemedia, bacteria that are strongly related to periodontal disease; and also to study the immunomudulatory mechanisms of all L. reuteri preparations in an invertebrate model (Galleria mellonella) and in periodontal ligament, osteoblastic cells and keratinocytes (in vitro). For antimicrobial analysis, the broth dilution test will be performed, with the combination of pathogenic bacteria with L. reuteri alive, dead or its supernatant. In G. mellonella, after infection with P. gingivalis, F. nucleatum, or P. intermedia and L. reuteri preparations, survival curve and hemocyte density will be analyzed. Periodontal ligament cells (hPdLF), keratinocytes (HaCaT) and osteoblastic cells (MG63) will be treated or not with preparations of L. reuteri and challenged with lipopolysaccharide (LPS). Then, levels of interleukin (IL)-1beta, IL-6, IL-17, IL-10, IL-12, tumor necrosis factor, matrix metalloproteinase (MMP)-9 and tissue inhibitor of MMP (TIMP)-1 will be evaluated by enzyme-linked immunosorbent assay. Nitric oxide production will be determined indirectly by the concentration of nitrite detected by Griess reagent. Levels of alkaline phosphatase and mineralization nodules will be evaluated in osteoblastic cells. The data will be analyzed with the appropriate statistical test, at 5% significance level. (AU)

Scientific publications
(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)
SANTOS, THAIS AGUIAR; SCORZONI, LILIANA; CORREIA, RAQUEL; JUNQUEIRA, JULIANA CAMPOS; ANBINDER, ANA LIA. Interaction between Lactobacillus reuteri and periodontopathogenic bacteria using in vitro and in vivo (G. mellonella) approaches. PATHOGENS AND DISEASE, v. 78, n. 8 DEC 2020. Web of Science Citations: 1.
GERALDO, BARBARA M. C.; BATALHA, MARIANNA N.; MILHAN, NOALA V. M.; ROSSONI, RODNEI D.; SCORZONI, LILIANA; ANBINDER, ANA LIA. Heat-killed Lactobacillus reuteri and cell-free culture supernatant have similar effects to viable probiotics during interaction with Porphyromonas gingivalis. JOURNAL OF PERIODONTAL RESEARCH, NOV 2019. Web of Science Citations: 0.

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