Scholarship 16/24808-5 - Biologia celular, Osteoclastogênese - BV FAPESP
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Study of the modulation of apical papilla cells by lipopolysaccharide and necrotic cells supernatant on in vitro osteoclastogenesis induction

Grant number: 16/24808-5
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Start date until: February 01, 2017
End date until: May 31, 2017
Field of knowledge:Health Sciences - Dentistry - Endodontics
Principal Investigator:Carla Renata Sipert
Grantee:Pamela Rocha Lopes de Almeida
Host Institution: Faculdade de Odontologia (FO). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Teeth with open apices accomplished by pulp necrosis present important structure commitment with short roots and thin dentin walls. At the same time, the development of periapical bone resorption areas is observed. The scientific literature has demonstrated mechanisms involved with the production of mineralized matrix by apical papilla cells, however evidences regarding the capacity of these cells in the modulation of bone resorption-related events are still lacking. Therefore, this project aims to evaluate the potential of apical papilla cells in the modulation of osteoclastogenesis in vitro. Primary cultures of apical papilla cells (n = 3) will be established based on an explant technique. Part of the cells will be primed with 1 µg/mL of Escherichia coli lipopolysaccharide (LPS) or necrotic cells supernatant at 1/10 fold serial dilution (SN) for 24 hours. The cells will be trypsinized and distributed at 24-wells plates. The activated-apical papilla cells or not will be kept with culture medium for 5 days and will be used for preparation of conditioned media. The quantification of IL-6, MCP-1, M-CSF, RANKL and OPG at cells supernatants will be performed by ELISA. The supernatant will be collected, centrifuged and stored. Peripheral blood mononuclear cells (PBMC) and CD14+ cells will be isolated and cultivated. Monocyte cultures will be kept in contact with the apical papilla cells-conditioned media for 21 days. The monocytes will be evaluated for immunostaining and activity of tartrate-resistant acid phosphatase (TRAP). Data will be analyzed by using one-way analysis of variance setting p < 0.05. (AU)

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