We quantitatively evaluated the adhesion of human osteoblasts on orthopedic metallic substrates (Ti6Al4V alloy) with various surface roughnesses at several times after inoculation and studied its correlation with qualitative changes in the expression of adhesion proteins and with parameters extensively describing the surface topographies. Cells were orientated in a parallel order on polished surfaces. This orientation was not affected by residual grooves after polishing. On sandblasted surfaces the cells never attained confluence and had a stellate shape, and the cell layer had no particular organization. Extracellular matrix (fibronectin, type I collagen, osteopontin) and cytoskeletal protein (actin, vinculin) orientation reflected the cell layer organization. In our experiment human osteoblasts expressed alpha3beta1 integrin but not alpha2beta1 integrin. In addition to currently analyzed roughness magnitude parameters, we calculated roughness organization parameters (fractal dimension parameters) of the substrates. We observed lower adhesion and proliferation on less organized surfaces (i.e., sandblasted ones). The significant statistical correlation observed between fractal dimension parameters (describing surface roughness organization) and cell parameters adds a new concept to the studies of substratum roughness influence on cell behavior. An attempt at modelization of the cell-surface interaction was made that includes the influence of fractal dimensions parameters.
We quantitatively evaluated the adhesion of human osteoblasts on orthopedic metallic substrates (Ti6Al4V alloy) with various surface roughnesses at several times after inoculation and studied its correlation with qualitative changes in the expression of adhesion proteins and with parameters extensively describing the surface topographies. Cells were orientated in a parallel order on polished surfaces. This orientation was not affected by residual grooves after polishing. On sandblasted surfaces the cells never attained confluence and had a stellate shape, and the cell layer had no particular organization. Extracellular matrix (fibronectin, type I collagen, osteopontin) and cytoskeletal protein (actin, vinculin) orientation reflected the cell layer organization. In our experiment human osteoblasts expressed ␣31 integrin but not ␣21 integrin. In addition to currently ana-lyzed roughness magnitude parameters, we calculated roughness organization parameters (fractal dimension parameters) of the substrates. We observed lower adhesion and proliferation on less organized surfaces (i.e., sandblasted ones). The significant statistical correlation observed between fractal dimension parameters (describing surface roughness organization) and cell parameters adds a new concept to the studies of substratum roughness influence on cell behavior. An attempt at modelization of the cell-surface interaction was made that includes the influence of fractal dimensions parameters.
Various surface treatments have been developed to increase the clinical performance of titanium-based implants. Many in vitro tests have been carried out on substrates with varied surface topography for a complete understanding of osteoblasts. In previous research, we made the observation that surface roughness must be taken into account, not only in terms of amplitude but also in terms of organization. In this study, we tested the adhesion and proliferation of human primary osteoblasts on grooved titanium surfaces with various amplitudes and organizations of topography. The roughness was described at a scale above (macro-roughness) or below (micro-roughness) the cell size. We observed better orientation and proliferation of human osteoblasts on surfaces with a micro-roughness characterized by a lower Order (parameter describing the organization of topography) and by a higher Ra and Rz (parameters describing the amplitude of topography). It appears that cultured human osteoblasts prefer surfaces with relatively high micro-roughness amplitude and with a low level of repeatability.
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