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Proteomic analysis of periodontal tissues with mechanical orthodontic force on health or obesity - in vivo and in vitro study

Grant number: 18/25811-5
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): April 01, 2019
Status:Discontinued
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Joni Augusto Cirelli
Grantee:Camila Chierici Marcantonio
Home Institution: Faculdade de Odontologia (FOAr). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

The aim of this research project is to evaluate in vivo and in vitro the effect of obesity on periodontal tissue remodeling induced by mechanical orthodontic force. For the in vivo study, the samples collected in the student Master's research (2016/00732- 0) will be used. Proteomics analysis of the animals periodontal ligament induced by orthodontic movement and submitted or not to obesity induced will be microdissected through laser capture microdissection (LCM). These samples will be used and to liquid chromatrography-high resolution mass spectrometry (LC-MS/MS) analysis. Immunohistochemical analysis will be used to confirm the detection and localization of the six more relevant proteins previous found by proteomics analysis. Furthermore, the gene expression of these proteins will be evaluated on the animals gingival tissue by RT-qPCR. For the in vitro study, a human immortalized periodontal ligament-derived cell line will be use. These cells will be exposed to a static tensile stain (STS) on a high magnitude (STSH), with the association or not of Adiponectin and NAMPT stimulation. The gene expression of six more relevant proteins previous found on in vivo study will be evaluated by RT-qPCR. These proteins will be confirmed by ELISA and intracellular signaling and nuclear translocations induced will be analyzed by immunofluorescence. All data will be evaluated, and then subjected to normality test. Then, appropriate statistical tests for comparisons between groups will be used. For all tests, a 5% significance level will be adopted.