To develop an endodontic cement using bovine bone-derived hydroxyapatite (BHA), Portland cement (PC), and a radiopacifier. Methods: BHA was manufactured from waste bovine bone and milled to form a powder. The cements were developed by the addition of BHA (10%/20%/30%/40% wt), 35% wt, zirconium oxide (radiopacifier) to Portland cement (PC). A 10% nanohydroxyapatite (NHA) cement containing PC and a radiopacifier, and a cement containing PC (PC65) and a radiopacifier were also manufactured as controls. The cements were characterised to evaluate their compressive strength, setting time, radiopacity, solubility, and pH. The biocompatibility was assessed using Saos-2 cells where ProRoot MTA acted as the control. Compressive strength, solubility and pH were evaluated over a 4-week curing period. Results: The compressive strength (CS) of all cements increased with the extended curing times, with a significant CS increase in all groups from day 1 to day 28. The BHA 10% exhibited significantly higher CS compared with the other cements at all time points investigated. The BHA 10% and 20% groups exhibited significantly longer setting times than BHA 30%, 40% and PC65. The addition of ZrO2 in concentrations above 20% wt and Ta2O5 at 30% wt resulted in a radiopacity equal to, or exceeding that of, ProRoot MTA. The experimental cements exhibited relatively low cytotoxicity, solubility and an alkaline pH. Conclusions: The addition of 10% and 20% BHA to an experimental PC-based cement containing 35% ZrO2 improved the material’s mechanical strength while enabling similar radiopacity and biocompatibility to ProRoot MTA. Although BHA is a cost-effective, biomimetic additive that can improve the properties of calcium silicate endodontic cements, further studies are now warranted to determine its clinical potential.