Implants are widely used in the human body for the replacement of affected bones. Fatigue failure is one of the serious concerns for implants. Therefore, understanding of the underlying mechanism leading to fatigue failure is important for the longevity of biomaterial implants. In this paper, the fracture toughness and fatigue crack growth of titanium alloy biomaterial Ti-27Nb has been experimentally investigated. The Ti-27Nb material is tested for fatigue crack growth in different environmental conditions representing the ambient and in vitro environments for 504 hours and 816 hours, respectively. Fractography of the tested specimen is conducted using Scanning Electron Microscope (SEM). The results of the fatigue crack growth propagation of the ambient and in vitro samples are similar in the Paris crack growth region. However, in the threshold region, the crack growth rate is higher for the Simulated Body Fluid (SBF) treated specimen. The fracture surface morphology of in vitro samples shows brittle fracture as compared to ambient specimens with significant plasticity and striations marks. It is proposed that a similar investigation may be conducted with specimens treated in SBF for prolonged periods to further ascertain the findings of this study.