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Doctoral Thesis
DOI
https://doi.org/10.11606/T.23.2021.tde-27102021-123034
Document
Author
Full name
Lorraine Perciliano de Faria
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2021
Supervisor
Committee
Chavez, Victor Elias Arana (President)
Cespedes, Maria Cristina Manzanares
Ervolino, Edilson
Ferraz, Emanuela Prado
Title in Portuguese
Efeito do alendronato, da dexametasona e da associação de ambos sobre a osteoclastogênese: um estudo in vitro
Keywords in Portuguese
Bisfosfonato
Cultivo celular
Glicocorticoide
Osteoclasto
Abstract in Portuguese
Os osteoclastos são células multinucleadas com a função de degradar e reabsorver o tecido ósseo. Medicamentos como o alendronato (um tipo de bisfosfonato nitrogenado) e a dexametasona (glicocorticoide), podem interferir na fisiologia das células clásticas. Enquanto o alendronato (ALN) inativa o osteoclasto, agindo primordialmente sobre o citoesqueleto desta célula, a dexametasona (DEX) pode promover aumento na atividade dessa célula. O objetivo deste estudo foi avaliar os efeitos do ALN, da DEX e da combinação de ambos sobre a osteoclastogênese e ativação desta célula. Primeiramente foi estabelecido protocolo de remoção de smear layer para os discos de osso bovino e na sequência, foi realizada análise em espectrofotômetro para avaliar a concentração de ALN absorvida pelo osso. O substrato ósseo foi capaz de absorver completamente o ALN da solução nas concentrações de 10 e 100 M. O cultivo de osteoclastos foi feito a partir das células da medula óssea de camundongos e estimulados com 1,25 dihidroxivitamina D3 e através de osteoclastos obtidos a partir de células Raw 264.7 estimulados com RANKL. As células foram cultivadas sobre substrato ósseo previamente tratado com ALN e tratadas com DEX a 1 M. Conclui-se que o tratamento com ALN a 10 M não foi capaz de inibir completamente a reabsorção óssea, seja administrado sozinho ou com a DEX. A DEX promoveu aumento na expressão gênica RANKL e redução de OPG, mesmo quando administrada conjuntamente com ALN. Quando utilizado na concentração de 100 M, o ALN reduziu a quantidade de anéis de actina dos osteoclastos e promoveu significativa diminuição na liberação de EVs nestas células.
Title in English
Effect of alendronate, dexamethasone and the association of both on osteoclastogenesis: an in vitro study
Keywords in English
Bisphosphonate
Cell culture
Glucocorticoid
Osteoclast
Abstract in English
Osteoclasts are multinucleated cells which degrade and reabsorb bone tissue. Drugs such as alendronate (nitrogen-containing type of bisphosphonate) and dexamethasone (glucocorticoid), have several effects on osteoclasts. While alendronate (ALN) inactivates the osteoclast, acting primarily on the cytoskeleton of this cell, dexamethasone (DEX) may promote an increase in this cell activity. The aim of this study was to evaluate the effects of ALN, DEX and a combination of both on osteoclastogenesis and activation of this cell. First, a smear layer removal protocol for bovine bone disks was established, and then a spectrophotometric analysis to assess the concentration of ALN absorbed by the bone. The bone substrate was able to completely absorb the ALN with solutions of 10 and 100 M. Osteoclast culture was obtained from mouse bone marrow cells and stimulated with 1.25 dihydroxyvitamin D3 and through osteoclasts from Raw 264.7 cells stimulated with RANKL. The cells were cultivated on bone slices treated with ALN and DEX at 1 M. It was concluded that treatment with ALN at 10 M did not completely inhibit bone resorption, whether administered alone or with DEX. DEX promoted increased expression of the RANKL and reduced OPG, even when administered with ALN. When osteoclasts were treated with 100 M ALN, a significant decrease in the formation of actin rings was found and reduced release of EVs.
 
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Publishing Date
2022-01-18
 
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