| Grant number: | 16/11450-5 |
| Support Opportunities: | Scholarships in Brazil - Post-Doctoral |
| Start date: | September 01, 2016 |
| End date: | December 31, 2019 |
| Field of knowledge: | Health Sciences - Dentistry |
| Principal Investigator: | Carlos Ferreira dos Santos |
| Grantee: | Bella Luna Colombini Ishikiriama |
| Host Institution: | Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil |
| Associated research grant: | 15/03965-2 - Role of the renin-angiotensin system in different oral inflammatory models: an experimental interdisciplinary and clinical approach, AP.TEM |
Abstract The dental pulp of deciduous teeth is a tissue that suffers important cellular and molecular alterations resulting from dental caries. Once known as a distinct tissue compared to permanent teeth dental pulps, studies clarifying the molecular mechanisms involved with these alterations deserve special attention. The aim of this study is to investigate components of the Renin Angiotensin System in molecular mechanisms involved with the modulation of the inflammatory process in deciduous dental pulp cells. Fibroblasts cultures will be established using an explant technique from dental pulp from deciduous teeth (n=3). Cells will be stimulated with increasing concentrations of lipoteichoic acid (LTA), lipopolysaccharide (LPS) and Interleukin(IL)-1² for detection of gene expression of Angiotensin receptor (AT1), Mas receptor, Angiotensin Converting Enzyme (ACE) and ACE2 besides Tumor Necrosis Factor(TNF)-± and Receptor Activator of NF-ºB (RANKL). Protein levels of AT1, Mas, ACE and ACE2 will be investigated by means of flow cytometry and RANKL and TNF-±, by enzyme linked immunosorbent assay (ELISA). The role of AT1, Mas, ACE and ACE2 on the production of pro- and anti-inflammatory mediators besides collagen I and ±-Smooth Muscle Actin (±-SMA) will be investigated by means of pharmacologic blockade and gene silencing. Lately, the cells will be stimulated with Angiotensin II (AngII) and Ang(1-7) for evaluation of the same parameters. The role of Nuclear Factor ºB (NF-ºB) activation and Transforming Growth Factor (TGF)-²1 receptor on the effects of AngII and Ang(1-7) will be investigated by means of pharmacological blockade. Finally, the modulation of the events observed by microRNAs will be investigated by means of the detection of these molecules by Reverse Transcription and quantitative Polimerase Chain Reaction (RT-qPCR) followed by gene silencing. Statistical analysis will be performed by means of Analysis of Variance (ANOVA) followed by Bonferroni´s correction test considering significant levels of p<0.05. | |
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