Objective: To evaluate the biological safety of manufactured heterologous deproteinized bone and to provide an experimental basis for clinical applications. Methods: Deproteinized bone ( 10 mm) and leaching liquor were made from pig ribs with a series of physical and chemical methods, then were evaluated through acute and subacute toxicity test, hemolysis test, pyrogen test,intracutaneous test, intramuscular implantation test and cytotoxity test.Results: No obvious toxicity, hemolysis, pyrogenic characteristics, skin irritation, inflammatory reaction after intramuscular implantation and cytotoxity were observed.Conclusions: The heterologous deproteinized bone has good biological safety and meets all the demands of scaffold material for tissue engineering.