Recent data indicate that the demineralization of vital bone promotes the consolidation of bone grafts and the proliferation and differentiation of pre - osteoblasts in culture, but the biological mechanisms involved in this process remain unclear. In rat calvaria, bone demineralization effects will be investigated for 1) the repair of onlay bone grafts whose contact surfaces shall be demineralized likewise; 2) the behavior of human osteoblasts cultured on demineralized bone surfaces in the same way as a) the cell morphology and surface area coverage by cells under scanning electron microscopy (SEM) ; b) the cellular synthetic activity by polymerase chain reaction in real-time (RT - PCR ) for expression of metalloproteinases (MMP - 2 , MMP -9), osteopontin (OPN), bone sialoprotein (BSP) the progressive ankylosis gene (ANKH), alcaline fosfatase, collagen tipe I and Runx-2; c) the cell ability to deposit calcium by colorimetric method. The demineralized surfaces will be characterized: on the composition through energy dispersive spectroscopy (EDS); for expression of bone morphogenetic proteins (BMPs) by confocal microscopy and for surface roughness by profilometry. Quantitative data collected from conventional optical microscopy, ¼CT, SEM, RT-PCR and colorimetry will be compared by ANOVA and Tukey's post-test and data analysis related to the bone surface will be compared by Student's t test, both with significance level of 95%. (AU)
Articles published in Agência FAPESP Newsletter about the research grant:
DO PRADO MANFREDI, GUSTAVO GONCALVES;
TOKUHARA, CINTIA KAZUKO;
SPADA DE CARVALHO, ERIKA BEATRIZ;
LIBOA-FILHO, PAULO NORONHA;
DAMANTE, CARLA ANDREOTTI;
PASSANEZI SANT'ANA, ADRIANA CAMPOS;
RAGGHIANTI ZANGRANDO, MARIANA SCHUTZER;
AGUIAR GREGHI, SEBASTIAO LUIS;
RUBO DE REZENDE, MARIA LUCIA.
Bone demineralization promotes superior spread of preosteoblast in culture.
MICROSCOPY RESEARCH AND TECHNIQUE,
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