Avaliação histomorfométrica, imuno-histoquímica e microtomográfica da neoformação óssea ao redor de implantes de titânio com diferentes superfícies instalados em áreas de defeitos ósseos preenchidos ou não com biomaterial
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2019-11-14
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Universidade Estadual Paulista (Unesp)
Resumo
O objetivo desse estudo foi avaliar a resposta óssea peri-implantar de implantes osseointegrados de titânio com dois tipos de tratamento de superfície, na presença ou não de substituto ósseo. Para tanto, foram utilizados 92 implantes dentários que foram divididos em quatro grupos de acordo com a superfície do implante e a presença de hidroxiapatita/beta-tricálcio fostato (HA/TCP) ou coágulo sanguíneo (BC) no defeito peri-implantar: (BC-A = implantes de superfície porosa e hidrofílica sem enxerto de substituto ósseo; HA/TCP-A = implantes de superfície porosa e hidrofílica com enxerto de substituto ósseo; BC-N = implantes de superfície porosa sem enxerto de substituto ósseo; HA/TCP-N = implantes de superfície porosa com enxerto de substituto ósseo). Os implantes foram instalados nas tíbias de 46 coelhos, sendo que cada animal recebeu dois implantes. Os animais foram submetidos a eutanásia em três diferentes períodos: 15 (n=10), 30 (n=10), e 60 (n=26). Após, as amostras foram submetidas a avaliação da qualidade óssea por meio das análises histológica por coloração Hematoxilina-Eosina, imuno-histoquímica para detecção dos anticorpos RUNX2, OPN, OCN, OPG, RANK-L e proteínas presentes na matriz óssea, utilizando imunoperoxidase para detecção e 3,3-diaminobenzidiana e de micro-tomografia computadorizada (micro-CT). Ainda, foi também realizada análise e avaliação histomorfométrica para quantificação do contato osso-implante (BIC) e a expressão gênica foi verificada por meio dos marcadores Runx2 e BSP, por meio da reação da transcriptase reversa, seguida de reação em cadeia da polimerase (RT-PCR). Os dados foram tabulados e submetidos a análise estatística por meio dos testes de Shapiro-Wilk, Kolgomorov-Smirnov e ANOVA. Em todos os períodos, o grupo HA/TCP-A apresentou uma porcentagem maior de neoformação óssea. No entanto, não houve diferenças estatisticamente significante para a neoformação óssea entre os grupos (p <0,05). Para os dois tipos de superfície, a presença do substituto ósseo apresentou valores mais altos em todos os parâmetros avaliados da micro-CT, exceto no parâmetro superfície/volume ósseo. A análise descritiva da imuno-histoquímica apresentou resultados semelhantes entre os grupos. Além disso, as superfícies porosas e hidrofílicas apresentaram uma expressão mais elevada de RUNX2. O grupo BC-A apresentou o maior valor médio do BIC (46,43%). As superfícies porosas e hidrofílicas também apresentaram melhores resultados de BIC quando comparadas à superfície porosa nas duas condições experimentais. Não foram encontradas diferenças estatisticamente significantes entre todos os grupos (p <0,05). Esses resultados sugerem que a superfície porosa e hidrofílica testada pode melhorar e acelerar a formação óssea, bem como a expressão de proteínas que desempenham papel importante na atividade osteogênica. E na presença de substituto ósseo, esta pode acelerar a formação de tecido ósseo peri-implantar. No entanto, após a osseointegração, ambas superfícies testadas tem comportamento similar tanto na presença quanto na ausência dos substitutos ósseos.
The aim of this study was to evaluate the peri-implant bone response of osseointegrated titanium implants with two types of surface treatment, with or without bone substitute. Ninety-two titanium implants were divided into four groups according to implant surface and presence of hydroxyapatite/β-tricalcium phosphate (HA/TCP) or blood clot (BC) in the peri-implant defect: (BC-A = porous-hydrophilic surface implants without bone substitute material; HA/TCP-A = porous-hydrophilic surface implants with bone substitute material; BC-N = porous surface implants without bone substitute material; HA/TCP-N = porous surface implants with bone substitute material). The implants were installed in the tibias of 46 rabbits, and each animal received two implants. The animals were euthanized in three different periods: 15 (n=10), 30 (n=10), and 60 (n=26). After, the samples were submitted to bone quality evaluation by means of histological analysis by Hematoxylin-Eosin staining, immunohistochemical analysis for detection of antibodies: RUNX2, OPN, OCN, OPG, RANK-L and bone matrix proteins using immunoperoxidase for detection and 3,3- diaminobenzidian and computerized tomography (micro-CT). In addition, histomorphometric analysis was also performed to quantify bone-implant contact (BIC), and gene expression was verified by RUNX2 and BSP markers by means of reverse transcriptase – polymerase chain reaction (RT-PCR). The data were tabulated and submitted to statistical analysis using Shapiro-Wilk, KolgomorovSmirnov and ANOVA tests. In all periods, the HA/TCP-A group presented higher percentage of bone formation. However, there were no statistically significant differences for bone formation among all groups (p <0.05). For both types of surface, the presence of bone substitute material presented higher values in all evaluated micro-CT parameters, except in the bone surface/volume parameter. Immunohistochemistry descriptive analysis showed similar results between groups. In addition, porous and hydrophilic surfaces showed a higher expression of Runx2. The BC-A group presented the highest mean value of BIC (46.43%). The poroushydrophilic surfaces also showed better BIC results when compared to the porous surface in both experimental conditions. No statistically significant differences were found among all groups (p <0.05). These results suggest that porous-hydrophilic surface may improve and accelerate bone formation, as well as the expression of proteins that play an important role in osteogenic activity. And in the presence of bone substitute, it can accelerate the formation of periimplant bone tissue. However, after osseointegration, both surfaces have similar behavior both in the presence and absence of bone substitute materials. Keywords: Dental Implantantion, Endosseous. Heterografts. Osteogenesis.
The aim of this study was to evaluate the peri-implant bone response of osseointegrated titanium implants with two types of surface treatment, with or without bone substitute. Ninety-two titanium implants were divided into four groups according to implant surface and presence of hydroxyapatite/β-tricalcium phosphate (HA/TCP) or blood clot (BC) in the peri-implant defect: (BC-A = porous-hydrophilic surface implants without bone substitute material; HA/TCP-A = porous-hydrophilic surface implants with bone substitute material; BC-N = porous surface implants without bone substitute material; HA/TCP-N = porous surface implants with bone substitute material). The implants were installed in the tibias of 46 rabbits, and each animal received two implants. The animals were euthanized in three different periods: 15 (n=10), 30 (n=10), and 60 (n=26). After, the samples were submitted to bone quality evaluation by means of histological analysis by Hematoxylin-Eosin staining, immunohistochemical analysis for detection of antibodies: RUNX2, OPN, OCN, OPG, RANK-L and bone matrix proteins using immunoperoxidase for detection and 3,3- diaminobenzidian and computerized tomography (micro-CT). In addition, histomorphometric analysis was also performed to quantify bone-implant contact (BIC), and gene expression was verified by RUNX2 and BSP markers by means of reverse transcriptase – polymerase chain reaction (RT-PCR). The data were tabulated and submitted to statistical analysis using Shapiro-Wilk, KolgomorovSmirnov and ANOVA tests. In all periods, the HA/TCP-A group presented higher percentage of bone formation. However, there were no statistically significant differences for bone formation among all groups (p <0.05). For both types of surface, the presence of bone substitute material presented higher values in all evaluated micro-CT parameters, except in the bone surface/volume parameter. Immunohistochemistry descriptive analysis showed similar results between groups. In addition, porous and hydrophilic surfaces showed a higher expression of Runx2. The BC-A group presented the highest mean value of BIC (46.43%). The poroushydrophilic surfaces also showed better BIC results when compared to the porous surface in both experimental conditions. No statistically significant differences were found among all groups (p <0.05). These results suggest that porous-hydrophilic surface may improve and accelerate bone formation, as well as the expression of proteins that play an important role in osteogenic activity. And in the presence of bone substitute, it can accelerate the formation of periimplant bone tissue. However, after osseointegration, both surfaces have similar behavior both in the presence and absence of bone substitute materials. Keywords: Dental Implantantion, Endosseous. Heterografts. Osteogenesis.
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Palavras-chave
Implantação Dentária Endo-Óssea, Xenoenxertos, Osteogênese, Interface Osso-Implante