The role of Endocannabinoids in the modulation of ... - BV FAPESP
Advanced search
Start date
Betweenand


The role of Endocannabinoids in the modulation of stem cells of the Apical Papilla in vitro

Full text
Author(s):
Claudia Caroline Bosio Meneses
Total Authors: 1
Document type: Doctoral Thesis
Press: São Paulo.
Institution: Universidade de São Paulo (USP). Faculdade de Odontologia (FO/SDO)
Defense date:
Examining board members:
Carla Renata Sipert; Marinella Holzhausen Caldeira; Anibal Roberto Diogenes; Carlos Ferreira dos Santos
Advisor: Carla Renata Sipert
Abstract

The apical papilla is a source of primary odontoblasts, which play an important role during the root formation process. The initiation of an inflammatory process in the pulp can lead to pulp necrosis and the interruption of root formation. Among the mediators involved in inflammatory processes are the endocannabinoids (ECbs), anandamide (AEA), and 2-arachidonoylglycerol (2-AG), which can modulate different cell functions by activating cannabinoid receptors (CB) 1 and 2, and the transient receptor potential vanniloid 1 (TRPV-1). The aim of this study was to investigate the gene expression of the components of the endocannabinoid system (ECS) and TRPV-1 in the stem cells of the apical papilla (SCAPs) as well as the role of ECbs in the modulation of cell functions. SCAPs were isolated and stimulated using LPS to assess cell viability, differentiation, cytokine production, and the gene expression of the ECS components; these experiments represent phases 1 and 2 of this study. In phase 3, the SCAPs were treated with the TRPV-1 antagonist capsazepine (CPZ) and LPS. In phase 4, SCAPs were activated using exogenous ECbs, AEA, and 2-AG to assess viability, cell differentiation, cytokine production, and the gene expression of mineralization markers. After seven days of treatment with LPS, there was no change in SCAP viability; however, there was an increase in cell differentiation in the group treated with 0.1µg/mL LPS, when compared with the control. A reduction in the production of osteoprotegerin (OPG) was observed in the groups stimulated using 0.1 and 10 µg/mL LPS along with an increase in MCP-1 (monocyte chemoattractant protein) / CCL-2 in the groups treated with 1 and 10 µg/mL LPS after 24 h. The gene expression of the components of ECS and TRPV-1 was observed in the SCAPs; however, expression of CB1 and CB2 was not observed. Cellular activation with LPS (10 µg/mL) and CPZ did not affect cell viability (72 h), differentiation (14 days), or the production of OPG and CCL-2 (24 h), compared with the control, regardless of treatment with CPZ. Exogenous ECbs promoted a reduction in cell viability in a dosedependent manner; however, AEA did not inhibit cell differentiation, regardless of the presence of CPZ. At high concentrations, 2-AG inhibited cell differentiation. No statistical difference was observed in the production of RANKL (receptor activator of nuclear kappa-B ligand) and tumor necrosis factor alpha (TNF-?) between any of the experimental groups. There was an increase in OPG production in the groups stimulated by AEA (with or without LPS) and in the group stimulated by 2-AG + LPS + CPZ after 24h. There was an increase in the production of IL-6 and CCL-2 in the groups treated with 2-AG; however, there was no change in the groups treated with AEA. A reduction in the expression of DMP-1 (dentin matrix acidic phosphoprotein-1) was observed in the group treated with AEA and an increased expression of DSPP was observed in the same group in 1 day, in comparison with the control. There was an increase in the expression of TGF-?1 (transforming growth factor ?-1) in the groups treated with AEA (with and without CPZ) within 1 day. Thus, we concluded that the main components of the ECS and TRPV-1 are expressed in the SCAPs. However, CB1 and CB2 are not expressed in the SCAPs. Additionally, at high concentrations, ECbs may affect SCAP viability and mineralization. (AU)

FAPESP's process: 17/01737-8 - Role of endocanabinoids on the modulation of human apical papilla cells in vitro
Grantee:Claudia Caroline Bosio Meneses
Support Opportunities: Scholarships in Brazil - Doctorate