One of the major limitations during titanium (Ti) implant osseointegration is the poor cellular interactions at the biointerface. In the present study, the combined effect of recombinant human Bone Morphogenetic Protein-2 (rhBMP-2) and nanopatterned Ti6Al4V fabricated with Directed irradiation synthesis (DIS) is investigated in vitro. This environmentally-friendly plasma uses ions to create self-organized nanostructures on the surfaces. Nanocones (≈36.7 nm in DIS 80°) and thinner nanowalls (≈16.5 nm in DIS 60°) were fabricated depending on DIS incidence angle and observed via scanning electron microscopy. All samples have a similar crystalline structure and wettability, except for sandblasted/acid-etched (SLA) and acid-etched/anodized (Anodized) samples which are more hydrophilic. Biological results revealed that the viability and adhesion properties (vinculin expression and cell spreading) of DIS 80° with BMP-2 were similar to those polished with BMP-2, yet we observed more filopodia on DIS 80° (≈39 filopodia/cell) compared to the other samples (<30 filopodia/cell). BMP-2 increased alkaline phosphatase activity in all samples, tending to be higher in DIS 80°. Moreover, in the mineralization studies, DIS 80° with BMP-2 and Anodized with BMP-2 increased the formation of calcium deposits (>3.3 fold) compared to polished with BMP-2. Hence, this study shows there is a synergistic effect of BMP-2 and DIS surface modification in improving Ti biological properties which could be applied to Ti bone implants to treat bone disease.