The deposition of agglomerated hydroxyapatite (HAp) powders by low-pressure cold spray has been a topic of interest in recent years. Key parameters influencing the deposition of HAp powders include particle morphology and impact kinetic energy. This work examines the deposition of HAp powders on various metal surfaces to assess the impact of substrate properties on the formation of HAp deposits via cold spray. The substrates studied here encompass metals with varying hardness and thermal conductivities, including Al6061, Inconel alloy 625, AISI 316 stainless steel, H13 tool steel, Ti6Al4V, and AZ31 alloy. Single-track experiments offer insights into the initial interactions between HAp particles and different substrate surfaces. In this study, the results indicate that the ductility of the substrate may enhance HAp particle deposition only at the first deposition stages where substrate/particle interaction is the most critical factor for deposition. Features on the substrate associated with the first deposition sprayed layer include localized substrate deformation and the formation of clusters of HAp agglomerates, which aid in HAp deposition. Furthermore, after multiple spraying passes on the various metallic surfaces, deposition efficiency was significantly reduced when the build-up process of HAp coatings shifted from ceramic/metal to ceramic/ceramic interactions. Overall, this study achieved agglomerated HAp deposits with high deposition efficiencies (30–60%) through single-track experiments and resulted in the preparation of HAp coatings on various substrates with thickness values ranging from 24 to 53 µm. These coatings exhibited bioactive behavior in simulated body fluid.