Nanoscale surface modifications influence peri-implant cell fate decisions and implant loading generates local tissue deformation, both of which will invariably impact bone healing. The objective of this study is to determine how loading affects healing around implants with nanotopography. Implants with a nanoporous surface were placed in over-sized osteotomies in rat tibiae and held stable by a system that permits controlled loading. Three regimens were applied: (a) no loading, (b) one daily loading session with a force of 1.5N, and (c) two such daily sessions. At 7 days post implantation, animals were sacrificed for histomorphometric and DNA microarray analyses. Implants subjected to no loading or only one daily loading session achieved high bone-implant contact (BIC), bone-implant distance (BID) and bone formation area near the implant (BFAt) values, while those subjected to two daily loading sessions showed less BFAt and BIC and more BID. Gene expression profiles differed between all groups mainly in unidentified genes, and no modulation of genes associated with inflammatory pathways was detected. These results indicate that implants with nanotopography can achieve a high level of bone formation even under micromotion and limit the inflammatory response to the implant surface.

This study evaluated the impact of a single variation in the etching time of H₂SO₄/H₂O₂-treated titanium (Ti) surfaces on the adsorption of growth and differentiation factor-5 (GDF-5) and their effects on the acquisition of the osteogenic phenotype in vitro. Rat primary calvarial osteogenic cells were grown for up to 14 days on the following Ti surfaces: (1) 30 min: nanotopography obtained with a 1:1 mixture of H₂SO₄/H₂O₂ for 30 min (control); (2) 30 min + GDF-5: a 30 min-etched Ti sample adsorbed with recombinant human (rh) GDF-5; (3) 4 h: nanotopography obtained with a 1:1 mixture of H₂SO₄/H₂O₂ for 4 h (control); (4) 4 h + GDF-5: a 4 h-etched Ti sample adsorbed with rhGDF-5. The GDF-5 adsorption procedure was carried out on the day before cell plating using 200 ng/mL rhGDF-5 overnight at 4 °C. The 30 min- and 4 h-etched Ti samples exhibited a high hydrophilic network of nanopits with a tendency towards larger nanopits for the 4 h group, which corresponded to an enhanced GDF-5 adsorption. For both etching times, coating with GDF-5 resulted in less hydrophilic surfaces that supported (1) a reduction in the proportion of spread cells and an enhanced extracellular osteopontin labeling at early time points of culture, and (2) increased alkaline phosphatase activity preceding an enhanced mineralized matrix formation compared with controls, with a tendency towards higher osteogenic activity for the 4 h + GDF-5 group. In conclusion, the osteogenic potential induced by the GDF-5 coating can be tailored by subtle changes in the nanotopographic characteristics of Ti surfaces.