Proliferation, Behavior, and Differentiation of Osteoblasts on Surfaces of Different Microroughness

Objectives: Titanium surface roughness is recognized as an important parameter influencing osseointegration. However, studies concerning the effect of well-defined surface topographies of titanium surfaces on osteoblasts have been limited in scope. In the present study we have investigated how Ti surfaces of different micrometer-scale roughness influence proliferation, migration, and differentiation of osteoblasts in-vitro.

Methods: Titanium replicas with surface roughnesses (Ra) of approximately 0, 1, 2, and 4μm were produced and MG-63 osteoblasts were cultured on these surfaces for up to 5 days. The effect of surface micrometer-scale roughness on proliferation, migration in time-lapse microscopy experiments, as well as the expression of alkaline phosphatase, osteocalcin, vascular-endothelial growth factor (VEGF), osteoprotegerin (OPG), and receptor activator of nuclear factor kappa-B ligand (RANKL) were investigated.

Results: Proliferation of MG-63 cells was found to decrease gradually with increasing surface roughness. However, the highest expression of alkaline phosphatase, osteocalcin and VEGF was observed on surfaces with Ra values of approximately 1 and 2μm. Further increase in surface roughness resulted in decreased expression of all investigated parameters. The cell migration speed measured in time-lapse microscopy experiments was significantly lower on surfaces with a Ra value of about 4μm, compared to those with lower roughness. No significant effect of surface roughness on the expression of OPG and RANKL was observed.

Significance: Thus, surfaces with intermediate Ra roughness values of 1-2μm seem to be optimal for osteoblast differentiation. Neither proliferation nor differentiation of osteoblasts appears to be supported by surfaces with higher or lower Ra values.