To evaluate the biocompatibility of a new nano TCP/gelatin/velvet antler polypeptide material.The nano TCP/gelatin/velvet antler polypeptide material was prepared, and the amorphous was observed by scanning electron microscope. L929 and NIH/3T3 cell lines were cultured conventionally. Acute toxicity test, hemolysis test, cell proliferation and cytotoxicity test were used to evaluate the biocompatibility of the material.The composite microsphere material was about 10 microm in diameter and had good spherical geometry, high monodispersity with nanometer size holes on the surface. Toxic symptoms such as hyperspasmia, palsy and death did not appear during the observing stage in acute toxicity test. Maximum hemolysis rate of the material was less than 5% which met the requirement of hemolysis test standard as a medical material. Different concentrations of the materials leaching liquor could enhance the proliferation of NIH/3T3 cells, which showed the good biologic activity. Toxicity grade was 0, and the material was no cytotoxic.Nano TCP/gelatin/velvet antler polypeptide material has good biocompatibility.