Purpose: Deproteinized bovine bone substitutes are commonly used in dental regenerative surgery for treatment of alveolar defects. In this study, three different bovine bone minerals - OCS-B (NIBEC, Seoul, Korea), Bio-Oss (Geistlich - Pharma, Switzerland), Osteograft/N - 300 (OGN, Dentsply Friadent Ceramed. TN, USA) - were investigated to analyze the basic characteristics of commercially available bone substitutes. Methods: Their physicochemical properties were evaluated by scanning electron microscopy, energy dispersive X-ray spectrometer (EDS), surface area analysis, and Kjeldahl protein analysis. Cell proliferation and alkaline phosphatase (ALP) activity of human osteosarcoma cells on different bovine bone minerals were evaluated. Results: Three kinds of bone substitutes displayed different surface properties. Ca/P ratio of OCS - B shown to be lower than other two bovine bone minerals in EDS analysis. Bio-Oss had wider surface area and lower amount of residual protein than OCS - B and OGN. In addition Bio - Oss was proved to have lower cell proliferation and ALP activity due to lots of residual micro particles, compared with OCS - B and OGN. Conclusions: Based on the results of this study, three bovine bone minerals that produced by similar methods appear to have different property and characteristics. It is suggested that detailed studies and quality management is needed in operations for dental use and its biological effects on new bone formation.