The management of large amounts of bio-wastes, such as bovine femurs from kitchens and slaughterhouses, has long been a challenging issue. However, through the utilization of a hydrothermal process, it is possible to transform these bio-wastes into valuable products. In this study, we focused on extracting hydroxyapatite (HAp), the primary inorganic component of bovine femurs, for potential use in bone tissue engineering scaffolds. By subjecting the femurs to hydrothermal treatment at varying times and solvents, we successfully decomposed and removed the organic matter present, resulting in the extraction of HAp. To comprehensively evaluate the properties of the extracted HAp, we employed several characterization techniques that provided valuable insights into the structure, morphology, and elemental composition of the extracted HAp. Furthermore, we conducted a Cell Counting Kit-8 assay, which confirmed the favorable biocompatibility of the extracted HAp. Overall, this study highlights the potential of hydrothermal treatment as an environmentally friendly and cost-effective method for handling bio-waste, specifically bovine femurs. The extracted HAp exhibits promising characteristics, making it suitable for a wide range of biomedical applications. This research contributes to the sustainable utilization of bio-waste and underscores the importance of resourceful exploitation for environmental protection. Waste bovine femur bone represents a valuable resource with rich hydroxyapatite (HAp) content, making it an attractive candidate for recycling purposes. HAp holds significant importance in the biomedical field due to its remarkable biological activity and other favorable properties. In this article, HAp was extracted from bovine femur waste by a hydrothermal method. The study extensively investigated the properties of HAp under varying reaction conditions, including particle size, morphology, and the presence of trace ions. The results from cellular experiments demonstrated the immense potential of the extracted HAp as a valuable biomaterial for various applications. This work opens up new avenues for utilizing waste bovine femur bone and contributes to the advancement of biomedical materials research.