Commercially pure titanium (CP-Ti) is a human-implant metal material commonly used for cardiovascular scaffolds and dental implants in the medical field. This is because CP-Ti has better biocompatibility and corrosion resistance compared to other alloys such as titanium-aluminum-vanadium alloy (Ti-6Al-4V). However, the low strength properties of CP-Ti have limited its wider application (e.g., load-bearing components). On the contrary, Novel β titanium alloys possess higher strength and lower elastic modulus, which has led to the consideration of Ti-Nb based alloys for biomedical applications, while also taking into consideration their biocompatibility and other mechanical properties. Recently, laminated metal composites (LMCs) have attracted a lot of attention due to the excellent properties of the constituent alloys. Direct laser deposition (DLD) is an additive manufacturing technology that can be potentially used to manufacture LMCs. In this work, the DLD process was used to manufacture Ti/TNTZO LMC, and CP-Ti and TNTZO alloy powders were the raw materials. Subsequently, the microstructure, phase composition, mechanical properties, and in vitro bioactivity of the Ti/TNTZO LMCs were analyzed. The results demonstrated that high-density, crack-free Ti/TNTZO can be fabricated using the DLD process. Ti/TNTZO is mainly composed of α/α' and β phases. Transmitted Kikuchi diffraction maps showed the presence of α" martensite, but due to its low content, there were no relevant peaks in the X-ray powder diffraction spectra. The hardness of the Ti region in the Ti/TNTZO increased due to the diffusion of alloy elements and refinement of the structure formed as a result of a faster cooling rate. However, for the TNTZO region, the hardness also increased due to the martensite transformation caused by the dilution of β-stabilizing elements compared with the TNTZO manufactured using the DLD process. In comparison with the CP-Ti and TNTZO made using the DLD process, the microstructure of the Ti/TNTZO multilayered materials was significantly different. The microstructure of Ti layers had coarse columnar grains and fine α/α' plates, and there was acicular martensite at the subgrain boundary of the TNTZO layers. As a result of the alloy elements diffusion, transition layer with a size of approximately 50 μm was found between the Ti layer and TNTZO layer. The tensile test results also showed that the multilayered materials have high yield strength and ultimate tensile strength. However, the presence of acicular martensite at the interface reduces the plasticity of the materials. Additionally, the Ti/TNTZO multilayered materials showed good ability to induce apatite formation after soaking in simulated body fluid for 14 d. Therefore, the results of this study showed that the Ti/TNTZO multilayered composites fabricated using the DLD process have potential application in the biomedical field.