This research evaluates the cellular response from Ti6Al4V scaffolds fabricated by hybrid additive manufacturing (AM) with the goal of improving bone tissue growth and biocompatibility of titanium implants. Since cellular adhesion and viability are subject to microstructure, hybrid AM was sought to dictate biological response by creating a dispersed and gradient microstructure within implants. The objective was to evaluate the effect of gradient microstructures within Ti6Al4V scaffolds on cellular metabolic activity. Gradient microstructure was incorporated within scaffolds by hybrid AM coupling directed energy deposition (DED) with interlayer milling. The cellular response from the scaffolds was evaluated using cell viability assays in two stages: the Ti6Al4V powder used for DED was first evaluated, followed by the fabricated scaffolds. Cell viability on hybrid AM scaffolds was compared with as–printed, i.e., DED-fabricated and annealed Ti6Al4V scaffolds. The cell viability assays demonstrated that Ti6Al4V powder was non-toxic as cells cultured with powder exhibited metabolic activity similar to cells in growth media without powder. The cell viability study on scaffolds indicated, on average, cell adhesion on hybrid AM scaffolds outperformed as–printed and annealed scaffolds. This study provides preliminary data demonstrating the use of hybrid AM to dictate cell activity. A larger sample set would be required in future work to understand the effects of grain refinement through coldworking on biological response.