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Role of osteoprotegerin on in vitro modulation of monocytes by apical papilla cells

Grant number: 22/14686-0
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): November 01, 2023
Effective date (End): October 31, 2027
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Fernando Neves Nogueira
Grantee:Letícia Martins Santos
Host Institution: Faculdade de Odontologia (FO). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

The resorption of mineralized tissues is a biological event present in challenging oral pathologies such as periodontal disease, apical periodontitis, and external root resorption. The role of mesenchymal stem cells in modulating bone metabolism is already known, but not the role of apical papilla cells in this context, especially through the effect of osteoprotegerin (OPG). The aim of this study is to investigate the role of OPG produced by SCAP in modulating apoptosis and monocyte differentiation through its binding to TRAIL and/or RANKL, under inflammatory conditions. SCAP will be pre-activated with lipopolysaccharide from Escherichia coli, and then plated. Cells will be analyzed for gene expression of OPG, TRAIL, and RANKL by RT-qPCR, and the supernatant will be collected for detection of OPG, TRAIL, IL-33, and RANKL by ELISA. Based on these findings, media conditioned with neutralizing antibodies to OPG will be prepared. Peripheral blood monocytes will be isolated and placed in culture. They will be treated with SCAP-anti-OPG or SCAP conditioned media, both collected from pre-activated or control cells and will be investigated for cell viability and apoptosis in the presence or absence of TRAIL and IL-33 neutralizing antibodies. Next, monocytes will be induced to differentiate into osteoclasts with the addition of M-CSF and RANKL in the presence of conditioned media for 21 days. The evaluation of the differentiation in osteoclasts will be done through the analysis of actin rings, detection of tartrate-resistant acid phosphatase (TRAP), and cathepsin K. The co-culture of SCAP and monocytes will be established to evaluate the ability of SCAP to modulate osteoclastogenesis through RANKL present in the cell membrane. The data will be analyzed statistically. (AU)

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