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Doctoral Thesis
DOI
Document
Author
Full name
Emanuel da Silva Rovai
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2018
Supervisor
Committee
Caldeira, Marinella Holzhausen (President)
Marques, Marcia Martins
Marco, Andrea Carvalho de
Villar, Cristina Cunha
Title in Portuguese
Efeito da ativação do receptor ativado por protease do tipo 1 (PAR1) sobre as atividades osteogênica e cementogênica de células mesenquimais do ligamento periodontal
Keywords in Portuguese
células do ligamento periodontal
potencial osteogênico
receptor do tipo 1 ativado por protease
Regeneração periodontal
trombina
Abstract in Portuguese
O receptor ativado por protease do tipo 1 (PAR1) foi o primeiro membro clonado da família de receptores acoplados à proteína G. Sua ativação tem sido associada ao reparo tecidual e cicatrização óssea. O objetivo do presente estudo foi avaliar o efeito da ativação do PAR1 nas atividades osteogênica e cementogênica de células mesenquimais do ligamento periodontal (CMLP). CMLP obtidas de 3 indivíduos foram cultivadas e tratadas com meio clonogênico (MC) ou meio osteogênico (MO) por 2, 7 e 14 dias. Depósitos de cálcio, concentração de cálcio (sobrenadante), atividade de fosfatase alcalina (ALP), proliferação celular, expressão gênica (qPCR) e níveis proteicos (ELISA) de fatores osteogênicos e cementogênicos foram avaliados na presença de trombina, agonista do PAR1 ou antagonista do PAR1. A ativação do PAR1 levou ao aumento da formação de depósitos de cálcio (p<0,05), o que foi associado ao aumento da concentração de cálcio (p<0,05), atividade da ALP (p<0,05) e proliferação celular (p<0,05). Além disso, os ensaios qPCR e ELISA mostraram que a ativação do PAR1 pode aumentar a expressão gênica de Runx2, OPG e CEMP1 (p<0,05) e níveis proteicos de Runx2 e OPG (p<0,05). Em conclusão, nossos resultados demonstram que a ativação de PAR1 aumenta as atividades osteogênica e cementogênica de CMLP.
Title in English
Effect of protease activated receptor type 1 (PAR1) activation on the osteogenic activity of periodontal ligament cells
Keywords in English
Osteogenic potential
periodontal ligament cells
Periodontal regeneration
Protease activated receptor type 1
Thrombin
Abstract in English
Protease activated receptor 1 (PAR1) was the first cloned member of the G protein-coupled receptor family. Its activation has been associated to tissue repair and bone healing. The aim of the present study was to evaluate the effect of PAR1 activation on the osteogenic and cementogenic activities of human periodontal ligament stem cells (HPLSC). HPLSC obtained from 3 subjects and treated with a control medium or with an osteogenic medium for 2, 7 and 14 days. Calcium deposits, calcium concentration (supernatant), alkaline phosphatase activity (ALP), cell proliferation and gene (qPCR) and protein expression (ELISA assay) of osteogenic and cementogenic factors were assessed in the presence of thrombin, PAR1 specific agonist peptide or PAR1 antagonist peptide. The activation of PAR1 led to increased formation of calcium deposits (p<0.05), which was associated to increased calcium concentration (p<0.05), ALP activity (p<0.05) and cell proliferation (p<0.05). Further, qPCR and ELISA assay showed that activation of PAR1 may increase gene expression of Runx2, OPG and CEMP1 (p<0.05) and protein levels of Runx2 and OPG (p<0.05). In conclusion, our results demonstrate that PAR1 activation increases osteogenic and cementogenic activities of HPLSC.
 
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Publishing Date
2019-05-16
 
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