Diferenciação de osteoblastos cultivados sobre superfícies de titânio modificados por irradiação com laser Yb: YAG pulsado de alta potencia
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Data
2013-04-06
Autores
Mariscal Muñoz, Eduardo [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
A osseointegração, requisito indispensável para o sucesso dos implantes dentários, é um processo lento, caracterizado, sequencialmente, pelas etapas de adesão, diferenciação e proliferação celular, bem como pela aposição e mineralização da matriz óssea depositada por osteoblastos. Acelerar o processo de osseointegração significa reduzir o tempo de espera para a aplicação segura de uma carga funcional sobre os implantes de titânio (Ti). Sabe-se que vários fatores, tal como a topografia da superfície do Ti, influenciam diretamente o processo de osseointegração. Assim, desde que foi demonstrado que alguns padrões específicos de superfície do Ti são capazes de bio-estimular osteoblastos, favorecendo e acelerando a osseointegração, diversas técnicas de baixo custo, rápida execução e altamente reproduzíveis, tem sido propostas para tratar a superfície dos implantes. Desta maneira, o objetivo da presente pesquisa foi avaliar a capacidade de superfícies de Ti, modificadas com laser pulsado de alta potência (L) ou usinadas (U), de estimular a diferenciação e maturação de células obtidas de calvária de camundongos cultivadas sobre elas. Para isto, foram realizados ensaios laboratoriais para determinar a atividade mineralizadora das células (coloração com vermelho de alizarina e fosfatase alcalina), o metabolismo (MTT assay) e morfologia celular (MEV). A fim de melhor caracterizar a diferenciação de osteoblastos, foi realizada a reação de polimerização em cadeia (PCR) quantitativa em tempo real para analisar a expressão, pelas células cultivadas sobre as superfícies de Ti, de genes que codificam os fatores de transcrição Runx2 e Sp7 e proteínas específicas da matriz extracelular mineralizada...
The osseointegration, which plays a fundamental role in the dental implantation success, is characterized by cell adhesion, differentiation, proliferation as well as deposition and mineralization of of bone matriz by osteoblasts. To make the osseointegration process faster means reducing the period to apply a safe functional stress on the titanium (Ti) implants. A number of factors, such as the topographical surface of Ti enhances the osseointegration process. Different Ti surface treatments capable of bio-estimulating osteoblasts to accelerate the osseointegration have been evaluated. Current studies have shown that osseointegration of Ti devices is enhanced by surface roughness. In this way, the aim of the present investigation was to assess the capacity of Ti surfaces irradiated with high potency laser (L) or polished (U) to stimulate the differentiation and maturation of cells obtained from calvarian bone of mouse. Then, laboratorial protocols to evaluate the mineralizing cell activity (alizarin red assay), cell metabolism (MTT assay) as well as the morphology (SEM) of cells cultured on the Ti surfaces were carried out. To better characterize the osteoblast differentiation, the real time qPCR for expression of genes that code to the transcription factors Runx2 and Sp7 were performed. Additionaly, this protocol was also used to assess specific proteins of extracellular matrix (Spp1, Alpl, e Col1a1). The numeric data were subjected to statistical analysis. Our data demonstrated that the Ti surface L improved the osteoblast maturation capacity of calvarial osteoblasts grown over this surface. Scanning electron microscopy (SEM) revealed spheres and protrusions created by laser treatment. Laser profilometry showed a disordered surface with micrometric and/to nanometric scales features (Ra = 10.57μm). When compared to polished... (Complete abstract click electronic access below)
The osseointegration, which plays a fundamental role in the dental implantation success, is characterized by cell adhesion, differentiation, proliferation as well as deposition and mineralization of of bone matriz by osteoblasts. To make the osseointegration process faster means reducing the period to apply a safe functional stress on the titanium (Ti) implants. A number of factors, such as the topographical surface of Ti enhances the osseointegration process. Different Ti surface treatments capable of bio-estimulating osteoblasts to accelerate the osseointegration have been evaluated. Current studies have shown that osseointegration of Ti devices is enhanced by surface roughness. In this way, the aim of the present investigation was to assess the capacity of Ti surfaces irradiated with high potency laser (L) or polished (U) to stimulate the differentiation and maturation of cells obtained from calvarian bone of mouse. Then, laboratorial protocols to evaluate the mineralizing cell activity (alizarin red assay), cell metabolism (MTT assay) as well as the morphology (SEM) of cells cultured on the Ti surfaces were carried out. To better characterize the osteoblast differentiation, the real time qPCR for expression of genes that code to the transcription factors Runx2 and Sp7 were performed. Additionaly, this protocol was also used to assess specific proteins of extracellular matrix (Spp1, Alpl, e Col1a1). The numeric data were subjected to statistical analysis. Our data demonstrated that the Ti surface L improved the osteoblast maturation capacity of calvarial osteoblasts grown over this surface. Scanning electron microscopy (SEM) revealed spheres and protrusions created by laser treatment. Laser profilometry showed a disordered surface with micrometric and/to nanometric scales features (Ra = 10.57μm). When compared to polished... (Complete abstract click electronic access below)
Descrição
Palavras-chave
Lasers, Titanio, Nanotecnologia, Titanium
Como citar
MARISCAL MUÑOZ, Eduardo. Diferenciação de osteoblastos cultivados sobre superfícies de titânio modificados por irradiação com laser Yb: YAG pulsado de alta potencia. 2013. 76 f. Dissertação (mestrado) Universidade Estadual Paulista Julio de Mesquita Filho, Faculdade de Odontologia de Araraquara, 2013.