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Doctoral Thesis
DOI
10.11606/T.42.2010.tde-25032010-150028
Document
Author
Full name
Daniela Scarabucci Janones
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2010
Supervisor
Committee
Chavez, Victor Elias Arana (President)
Acevedo, Ana Carolina
Oliveira, Newton Maciel
Santos, Marinilce Fagundes dos
Sousa, Suzana Cantanhede Orsini Machado de
Title in Portuguese
Estudo da presença de osteoaderina durante a ossificação intramembranosa e endocondral através de imunocitoquímica e Western Blotting
Keywords in Portuguese
Western Blotting
Imunocitoqímica
Mineralização Óssea
Ossificação Endocondral
Ossificação Intramebranosa
Osteoaderina
Abstract in Portuguese
A osteoaderina (OSAD) tem sido identificada nos tecidos mineralizados, porém, seu papel na mineralização óssea não está claro. Foi feita uma comparação do momento em que a OSAD aparece na ossificação intramembranosa e endocondral, em relação aos estágios iniciais de mineralização. O osso parietal de fetos de ratos Wistar com 17, 18 e 21 dias e o côndilo mandibular de ratos com 30 dias foram removidos. A expressão de OSAD foi analisada por imunocitoquímica e Western blotting. Nos dois tipos de ossificação, a imunomarcação foi detectada nos osteoblastos; porém, na matriz extracelular a OSAD apareceu somente na fase fibrilar de mineralização, mantendo-se constante posteriormente. A análise por Western blotting revelou que os fetos com 17 dias continham pouco menos OSAD que os de 18 dias, enquanto a imunorreatividade diminuía nos fetos com 21 dias. Os resultados sugerem que a OSAD tem um papel na mineralização da matriz, atuando, provavelmente como organizadora de seu arcabouço ou retendo o mineral, além de exercer atividades de adesão entre os componentes da matriz.
Title in English
Study of the osteoadherin presence during the intramembranous and endochondral ossification by immunocytochemistry and western blotting analysis
Keywords in English
Bone mineralization
Endochondral ossification
Immunocytochemistry
Intramebranous Ossification
Osteoadherin
Western Blotting
Abstract in English
Osteoadherin (OSAD) had been identified in mineralized tissues, but its specific role in mineralization remains unclear. The present study compared the appearance of OSAD at early stages of mineralization during both intramembranous and endochondral ossification. Parietal bone of 17, 18 and 21 days-old fetus and mandibular condyle of 30 days-old Wistar rats were removed. The expression of OSAD was analyzed by immunocytochemistry and Western blotting. In both types of ossification the labeling was uniformly distributed in the cytoplasm of osteoblasts but it only appeared in the mineralizing matrix when the fibrilar stage was taking place, remaining as a component of the mineralized bone matrix. Western blots revealed that 17-days-old embryos contained slightly less OSAD than 18- days-old fetus, while immunoreactivity was weak in 21 days-old fetus. The results suggest that OSAD plays a role in collagen fibril mineralization maybe by organizing the matrix assembly or by retaining the mineral into the matrix, besides exerting binding activities among its components.
 
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Publishing Date
2010-03-29
 
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