Tissue engineering aims to regenerate damaged tissue from the differentiation of osteogenic precursor cells upon scaffolds or bone graft materials. Development of bone graft materials has been thoroughly carried out to provide predictable and cost-effective bone regeneration. This paper aims to study the preparation of hydroxyapatite (HA) from natural bovine teeth as an alternative to existing bone graft materials in the field of dentistry. Hydroxyapatite (HA) powder was prepared from calcining and sintering bovine teeth at 735°C and 1200°C consecutively and were milled using High Energy Milling for 15 hours. X-Ray diffraction analysis (XRD) was run to confirm HA phase of bovine tooth powder after sintering and milling process. Particle Size Analyzer was used to measure the mean distribution of particle size of HA powder, and found that HA powder particle has an average size of 3.515 μm.