Study of the role of stem cells of the apical papilla on human peripheral blood monocytes migration in vitro

• Grant number: 23/01852-2
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: June 01, 2023
• End date: August 19, 2024
• Field of knowledge: Health Sciences - Dentistry - Endodontics
• Principal Investigator: Carla Renata Sipert
• Grantee: Alexandre Guimarães dos Santos
• Host Institution: Faculdade de Odontologia (FO). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Chemokines are cytokines that potentially attract distinct cell types trough chemotaxis. The CCL2 is the main chemokine involved at monocyte recruitment from peripheral blood to tissues; where, considering endodontic infections, these cells will differentiate into macrophages and osteoclasts. Based on the knowledge that stem cells of the apical papilla (SCAP) are an important source of CCL2, this study aims to investigate if SCAP culture supernatant is able to display chemotaxis on monocytes in vitro. For this purpose, SCAP conditioned medium will be prepared from primary SCAP culture, after determined the osteo/odontogenic differentiation potential of these cells. Then, the supernatant will be subjected to CCL2 quantitation by means of enzymatic immunoassay. The monocytes will be isolated from human peripheral blood and seeded at 8 um pore size transwell inserts that in turn will be placed over wells containing SCAP conditioned medium solely or seeded SCAP themselves. The cells that did not migrate will be gently removed from the top of the inserts and the remaining ones will be counted at light microscopy after fixed and stained with violet crystal. At predefined wells, CCL2 neutralizing antibody will be added or recombinant human CCL2 as positive control. Statistical analysis will be performed based on data normality standard. The significance level of 5% will be set as significant for group differences analysis.