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Master's Dissertation
DOI
10.11606/D.25.2013.tde-14082013-095031
Document
Author
Full name
Carolina Fávaro Francisconi
Institute/School/College
Knowledge Area
Date of Defense
Published
Bauru, 2013
Supervisor
Committee
Garlet, Gustavo Pompermaier (President)
Okamoto, Roberta
Oliveira, Rodrigo Cardoso de
Title in Portuguese
Caracterização morfométrica e molecar do papel da iNOS no processo de reparo ósseo alveolar em condições homeostáticas e infecciosas
Keywords in Portuguese
Alvéolo seco
Óxido nítrico
Tecido ósseo
Abstract in Portuguese
A óxido nítrico sintase induzível (iNOS) é uma enzima responsável pela síntese do óxido nítrico, envolvido na regulação de vários processos fisiológicos, com destaque para relevantes efeitos sobre o tecido ósseo. Entretanto, o papel da iNOS no reparo ósseo alveolar permanece pouco conhecido. Assim, o objetivo deste estudo foi caracterizar o papel da iNOS no processo de reparo ósseo alveolar pósexodontia em condições homeostáticas (controle) e infecciosas em camundongos. Para isso, foram utilizados animais das linhagens WT (C57Bl/6) e iNOSKO, divididos em 2 grupos, condição homeostática (exodontia do incisivo superior direito) e alveolite experimental (induzida pela inoculação de uma suspensão de cultura bacteriana), e analisados de acordo com os diferentes períodos experimentais (0, 7, 14 e 21 dias pós-exodontia), através da análise histomorfométrica e molecular. Na análise histomorfométrica, avaliou-se a densidade de volume das estruturas referentes ao coágulo sanguíneo, tecido conjuntivo e tecido ósseo. Na análise molecular, quantificou-se a expressão de mRNA codificando genes de marcadores do metabolismo ósseo; marcadores de osteoclastos; citocinas e quimiocinas, através de reações de RealTimePCR. A expressão de iNOS esteve presente durante todo o processo de reparo ósseo alveolar nos camundongos C57Bl/6, porém de maneira mais expressiva no período de 7 dias pós-exodontia, e se mostrou aumentada pela indução de alveolite. A análise histomorfométrica demonstrou discretas alterações no processo de reparo ósseo alveolar, na ausência de iNOS, tanto em condições homeostáticas quanto infecciosas. Em condições homeostáticas, a ausência de iNOS não impactou de forma significativa o processo de reparo ósseo alveolar pósexodontia, mas se mostrou associada à modulação de vasos sanguíneos. Já em condições infecciosas, a ausência de iNOS se mostrou associada à modulação de células inflamatórias, osteoblastos e osteoclastos. De forma geral, conclui-se que embora a enzima iNOS module certos aspectos do processo de reparo ósseo alveolar de forma pontual, sua ausência não interferiu de forma significativa nesse processo.
Title in English
Morphometric and molecular characterization of the role of iNOS in alveolar bone repair process in homeostatic and infectious conditions
Keywords in English
Bone and bones
Dry socket
Nitric oxide
Abstract in English
The inducible nitric oxide synthase (iNOS) is an enzyme responsible for the synthesis of nitric oxide, a reactive molecule involved in the regulation of several physiological processes, highlighting relevant effects on bone tissue. However, the role of iNOS in alveolar bone repair remains unknown. The aim of this study was to characterize the role of iNOS in alveolar bone healing process after dental extraction in homeostatic and infectious conditions in mice. With this aim, WT (C57Bl/6) and iNOSKO mice strains were divided into 2 groups, homeostatic condition (extraction of the upper right incisor) and experimental alveolitis (induced by inoculation of a suspension of bacterial culture), and analyzed according to the different experimental periods (0, 7, 14 and 21 days post-extraction), through molecular and histomorphometric analysis. In histomorphometric analysis, the volume density of structures related to blood clot, tissue and bone were evaluated. The molecular analysis quantified the expression of mRNA encoding genes of defined as bone metabolism markers; osteoclast markers, as well cytokines and chemokines through RealTimePCR reactions. The expression of iNOS was detected during the all process of the alveolar bone repair in C57Bl/6 mice, with an expression peak at 7 days post-extraction time point, and was significantly enhanced by alveolitis induction. Histomorphometric analysis showed small changes in alveolar bone repair process in the absence of iNOS, in both homeostatic as infectious conditions. Under homeostatic conditions, the absence of iNOS did not impact significantly the process of alveolar bone healing after dental extraction, but it was associated with modulation of blood vessels formation. In the infectious conditions scenario, the absence of iNOS was associated with modulation of inflammatory cells, osteoblasts and osteoclasts counts. In general, it is concluded that even the enzyme iNOS module some aspects of alveolar bone repair in a particular and punctual manner, their absence does not interfere significantly in the overall outcome of alveolar bone repair process.
 
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Publishing Date
2013-08-15
 
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