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The role of endocannabinoids in the modulation of apical papilla cells: an in vitro study

Grant number: 18/21869-9
Support type:Scholarships abroad - Research Internship - Doctorate
Effective date (Start): March 04, 2019
Effective date (End): September 03, 2019
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Carla Renata Sipert
Grantee:Claudia Caroline Bosio Meneses
Supervisor abroad: Anibal Roberto Diogenes
Home Institution: Faculdade de Odontologia (FO). Universidade de São Paulo (USP). São Paulo , SP, Brazil
Local de pesquisa : University of Texas Health Science Center at San Antonio (UTHSCSA), United States  
Associated to the scholarship:17/01737-8 - Role of endocanabinoids on the modulation of human apical papilla cells in vitro, BP.DR

Abstract

The apical papilla has an important role in root formation during dental development, acting as a source of stem cells that differentiate into the putative dentin-synthesizing cells, odontoblasts. Pulpal inflammation on immature teeth, resulting from caries or trauma, might lead to necrosis and the interruption of root formation. Among the inflammatory mediators potentially involved in this process, the endocannabinoids (ECbs) anandamide (AEA) and 2-arachydonoylglycerol (2-AG) might affect the behavior of apical papilla cells. Therefore, the aim of this study is to investigate the role of ECbs in the transient receptor potential vanilloid type 1 (TRPV1)-mediated molecular mechanisms involved in the phenotypic and functional alterations of apical papilla cells in vitro. Cultured stem cells of the apical papilla cells (SCAP) will be used between the 3rd and 5th passages. The cells will be pre-treated with capsazepine (a TRPV1 antagonist), followed by stimulation with AEA, 2-AG, or LPS for 1, 3, 7, and 14 days. Soon after, the cells will be submitted to a viability assay, the determination of cytokines and chemokines by ELISA, an Alizarin red S assay to analyze calcium deposition, and RT-qPCR to determine the cell differentiation profile. Statistical analysis will be performed using a one-way analysis of variance (ANOVA), followed by Tukey's post-test or Student's t-test. Significant values will be set at p<0.05.