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Abstract
Bone formation around implants is influenced by surface geometry. Since cell/matrix/substrate
interactions associated with cell signaling occur in the nanoscale dimension, we have
evaluated the influence of nanotexturing of titanium-based surfaces on the expression
of matrix proteins by cultured osteogenic cells at initial time points. Cells were
obtained by enzymatic digestion of newborn rat calvaria and grown on titanium and
titanium alloy discs with nanotextured or machined surfaces, and on glass coverslips
for periods of 6 h, 1 day, and 3 days, under standard culture conditions. Cultures
were processed for single or dual immunolabeling with monoclonal and/or polyclonal
antibodies against bone sialoprotein (BSP), fibronectin (FN), osteopontin (OPN), type-I
pro-collagen, or tubulin, followed by corresponding fluorophore-conjugated secondary
antibodies. Some samples were processed for scanning electron microscope analysis
of morphology and immunogold labeling. After 6 h, nanotextured surfaces exhibited
up to a nine-fold increase in the proportion of cells with peripheral OPN labeling.
At day 3, the proportion of OPN and BSP labeled cells was higher, and the intensity
of immunoreactivity dramatically increased. No significant differences were observed
in the expression pattern and the proportion of cells immunoreactive for FN or type-I
pro-collagen. Our results demonstrate that nanotexturing of titanium-based surfaces
upregulates the early expression of BSP and OPN in osteogenic cell cultures.