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Master's Dissertation
DOI
https://doi.org/10.11606/D.17.2016.tde-29052015-110800
Document
Author
Full name
Miliane Gonçalves Gonzaga
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2014
Supervisor
Committee
Issa, João Paulo Mardegan (President)
Ana, Lauro Wichert
Title in Portuguese
Estudo qualitativo e quantitativo da interação entre a proteína morfogenética rhBMP-2 e diferentes tipos de enxertos ósseos
Keywords in Portuguese
Enxerto
Histomorfometria
Reparo ósseo
rhBMP-2
Abstract in Portuguese
As proteínas com potencial osteoindutor, com destaque a rhBMP-2, possuem amplos efeitos sobre o crescimento e a diferenciação celular. Este estudo, teve o objetivo avaliar a relação entre diferentes enxertos ósseos e a rhBMP-2 no reparo de defeitos ósseos críticos em calvárias de ratos. Foram utilizados 56 ratos Wistar albinos, machos, aproximadamente 250g, com um período de espera de 6 semanas desde a criação do defeito até o sacrifício, divididos em 8 grupos (n=7): enxerto autólogo (AUT); enxerto homólogo (HOM); enxerto heterólogo (HET); enxerto autólogo e 5 g rhBMP-2 (AUT BMP); enxerto homólogo e 5 g rhBMP-2 (HOM BMP); enxerto heterólogo e 5 g rhBMP-2 (HET BMP); 5 g rhBMP-2 (BMP) e defeito ósseo crítico apenas (DOC). Os resultados foram avaliados estatisticamente pelo teste ANOVA e pós-teste de TUKEY com múltiplas comparações (nível de significância de p<0,05), após o emprego de metodologias de análise histológica qualitativa, histomorfometria e micro CT. Na quantificação do volume ósseo neoformado, todos os grupos obtiveram melhores resultados que DOC com diferença significante. Os grupos HOM e HET apresentaram diferença significante de AUT BMP, HOM BMP e HET BMP que tiveram maior formação óssea enquanto que AUT apresentou diferença significante de HOM BMP e HET BMP. O grupo BMP não teve diferença significante em relação a AUT, HOM, HET, AUT BMP, HOM BMP e HET BMP. Os grupos AUT, HOM e HET não tiveram diferença significante entre si da mesma forma que os grupos AUT BMP, HOM BMP e HET BMP também não o tiveram. Quanto à quantificação de osteoclastos, não houve diferença significante entre os grupos; porém, HET BMP teve mais células osteoblásticas que DOC com diferença significante. Considerando os osteócitos, DOC e AUT apresentaram poucas células. Assim, DOC teve diferença significante de BMP, HET, HOM BMP e HET BMP e AUT teve diferença significante de BMP, HET, HOM BMP e HET BMP. Da mesma forma, AUT BMP teve diferença significante em relação a HET que apresentou mais células. Na quantificação de fibras colágenas, não houve diferença significante entre os grupos. A neoformação óssea progrediu das bordas para o centro do defeito e no grupo BMP o osso neoformado ocupou mais de 2/3 do defeito. No grupo DOC, foi constatada uma faixa estreita de tecido ósseo neoformado nas bordas e o restante do defeito foi preenchido por tecido conjuntivo. Nas imagens de micro CT, todos os grupos apresentaram maior formação óssea que o grupo DOC; sendo que AUT superou HOM e HET. Além disso, os diferentes enxertos associados à rhBMP-2 apresentaram maior formação óssea que os respectivos enxertos usados sem a proteína, com destaque para AUT BMP. Diante disso, foi possível concluir que a rhBMP-2 auxiliou o reparo ósseo quando administrada isoladamente ou associada aos diferentes enxertos ósseos, conferindo neste caso melhores resultados
Title in English
Qualitative and quantitative study of the interaction between bone morphogenetic protein rhBMP-2 and different types of bone grafts
Keywords in English
Bone repair
Graft
Histomorphometry
rhBMP-2
Abstract in English
Proteins with osteoinductive potential, particularly the rhBMP-2, have great effects on growth and cell differentiation. This study aimed to evaluate the relationship between different bone grafts and rhBMP-2 on the repair of critical bone defects in rat skulls. Fifty six Wistar rats, male, approximately 250g of body weight were used, sacrificed six weeks after surgical experiment, and divided into 8 groups(n=7): autograft (AUT); homograft (HOM); xenograft (HET); autograft and 5 g rhBMP-2 (AUT BMP); homograft and 5 g rhBMP-2 (HOM BMP); xenograft and 5 g rhBMP-2 (HET BMP); 5 g rhBMP-2 (BMP) and critical bone defect only (DOC). The results were analyzed statistically by ANOVA and Tukey post-test with multiple comparisons (p<0.05 of significance level), after the use of qualitative methodologies, micro CT morphology and histological analysis. According to the quantification of newly formed bone volume, all groups had better results than DOC group with a significant difference. The HOM and HET groups showed significant difference than AUT BMP, HOM BMP and HET BMP, that had higher bone formation, while AUT group was significantly different from HOM BMP and HET BMP. The BMP group had no significant difference in relation to AUT, HOM, HET, AUT BMP, HOM BMP and HET BMP. The AUT, HOM and HET groups had no significant difference between them, and also the groups AUT BMP, HOM BMP and HET BMP between them. Regarding the quantification of osteoclasts, there was no significant difference between groups; however, HET BMP had more osteoblastic cells than DOC group, with significant difference. For osteocytes quantification, AUT and DOC showed few cells. Thus, DOC had significant difference in relation to BMP, HET, HOM BMP and HET BMP; and AUT had significant difference in relation to BMP, HET, HOM BMP and HET BMP. Likewise, AUT BMP had significant difference compared to HET group, which showed more cells. Regarding to collagen fibers quantification, there was no significant difference between groups. New bone formation progressed from the edges to the center of the defect and for BMP group the newly formed bone occupied more than two thirds of the defect. DOC group showed a narrow band of newly formed bone at the edges and the remainder of the defect was filled by connective tissue. For micro CT analysis, all groups showed greater bone formation than the DOC group, being greater in AUT group than HOM and HET. Furthermore, the various grafts associated with rhBMP-2 showed greater bone formation than the isolated grafts, highlighting the AUT BMP. Therefore, it was concluded that rhBMP-2 helped bone repair when administered alone or associated with different bone grafts, giving better results when combined to the grafts
 
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Publishing Date
2016-01-11
 
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