The DNA/protamine complex was prepared by a reaction between DNA and protamine sulfate solutions with stirring, and its cell viability, antibacterial effect and histopathological responses were examined. A water-insoluble white powder, DNA/protamine complex, with a porous structure was obtained. The molar binding ratio of the complex prepared from a solution containing equal amounts of DNA and protamine sulfate by weight was 0.038 and the efficiency of complex formation was 61%. In a cell culture test using MC-3T3-E1 mouse osteoblast cells, the complex showed less cytotoxicity than protamine sulfate alone and cell viabilities were more than 98%. A porous disk could be prepared easily and showed an antibacterial effect against Staphyrococcus aureus, Porphyromonas gingivalis and Prevotella intermedia in an antibacterial sensitivity test and a mild tissue response in vivo test. These results suggested that the DNA/protamine complex could be a useful biodegradable biomaterial with antibacterial effects.