Mechanical properties and biological evaluation of buffalo horn material were examined in this study. The effects of sampling position of buffalo horn on mechanical properties were investigated with uniaxial tension and micron indentation tests. Meanwhile, the variation of element contents in different parts of buffalo horn was determined with elemental analysis, and the microstructure of the horn was measured with scanning electron microscopy. In addition, biological evaluation of buffalo horn was studied with hemolytic test, erythrocyte morphology, platelet and erythrocyte count, and implantation into mouse. Results showed that the buffalo horn had good mechanical properties and mechanical characteristic values of it gradually increased along with the growth direction of the horn, which may be closely related to its microstructure and element content of C, N, and S in different parts of the buffalo horn. On the other hand, because the buffalo horn does not have toxicity, it therefore does not cause hemolysis of erythrocyte and has a good affinity with it. Buffalo horn has good histocompatibility but meanwhile it may induce the platelet adhesion and aggregation. Even so, it does not continue to rise to induce a large number of platelet to aggregate with resulting blood clotting. Therefore, the buffalo horn material has been proved to possess good blood compatibility according to the preliminary evaluation.