Bone graft materials are widely used at present because inadequate bone volume is usually found in implant patients. To determine the biocompatibility of a new grafting material, in vitro research is routinely performed before animal experiments and clinical testing. However, during in vitro experiments, bone material particles might move during testing, which could affect the accuracy of the results. MATERIALS AND METHODS: To evaluate the biocompatibility of new bone substitutes, Mega-oss and Mega-TCP were compared with Bio-Oss using osteoblast cells and osteoclast cells fixed with alginate gel. Cell morphology, viability, bone resorption, alkaline phosphatase (ALP) activity, and staining were tested to compare the biocompatibility differences in the performance of Mega-oss, Mega-TCP, and Bio-Oss. RESULTS: Cells spread better on Mega-oss and Mega-TCP than the round shape on Bio-Oss. The 3-(4,5-dimethyl-2-thiazolyl)-2,5diphenyl-2-H-tetrazolium bromide (MTT) results showed that Mega-oss, Mega-TCP, and sodium alginate had good viability. Meanwhile, Mega-oss and Mega-TCP had the same osteoblast differentiation ability as Bio-Oss. The resorption rates of Mega-TCP and Mega-oss were higher than those of Bio-Oss (24.4%, 15.3%, and 3.3%, respectively). CONCLUSION: Mega-oss and Mega-TCP might be useful alternative bone graft materials compared with Bio-Oss. In addition, fixing the materials with sodium alginate gel could be a new method for in vitro bone material experiments.