Abstract：[Objectives] To repair irregular bone defects, this study designed and prepared a composite artificial bone material using Acellular Bone Matrix/Calcium Phosphate / calcium sulfate hemihydrate (ACBM/CPC/CSH), and investigated its biocompatibility and in vivo osteogenic performance.[Methods] A composite artificial bone material, Acellular Bone Matrix/Calcium Phosphate/Sulfate (ACBM/CPC/CSH), was prepared using a physical mixing method, followed by comprehensive testing of the material''s biological safety, immunological characteristics, and osteogenic performance. ACBM/CPC/CSH material extracts were prepared for acute toxicity, subacute toxicity, pyrogenicity, and skin irritation experiments to observe the material''s biological safety. The experimental material was embedded in muscle tissues, and samples were collected on days 7, 14, and 21 for histopathological examination to observe the inflammatory response in the vicinity of the material. Flow cytometry analysis was conducted on day 14 to observe changes in CD4+ and CD8+ T lymphocyte content in blood and tissues, assessing the material''s immunological properties. Cell toxicity of the experimental material was evaluated using the CCK-8 assay. Utilizing a rat femoral medial condyle defect model, the study involved categorizing implanted materials into the ACBM/CPC/CSH experimental group, CPC/CSH control group, and a blank sham surgery group. X-ray examinations were conducted at 1, 2, and 4 weeks post-surgery, followed by Micro-CT examination at week 8 to comparatively study the osteogenic performance of different materials. [Results] (1) The extract of ACBM/CPC/CSH experimental material did not induce acute or subacute toxic reactions in mice, and there were no statistically significant differences in body weight changes among the groups (P > 0.05). Rabbit body temperatures remained within normal levels, with negative results in the pyrogenicity test. Continuous observation of skin reactions following subcutaneous injection showed no significant differences over multiple days. (2) Histopathological examination of the muscle tissue surrounding the implanted material revealed mild inflammation on day 7, which subsided by day 21. Flow cytometry analysis on day 14 indicated that the CD4+ and CD8+ T lymphocyte content showed no statistically significant differences compared to the blank group (P > 0.05). CCK-8 results demonstrated that the ACBM/CPC/CSH experimental material exhibited no cytotoxicity and displayed favorable immunological properties. (3) X-ray and Micro-CT examination results showed that with the passage of time, all groups exhibited varying degrees of bone defect repair. In the 8th week, Micro-CT observation revealed superior bone defect repair in the ACBM/CPC/CSH experimental group compared to the CPC/CSH group, and the bone material completely degraded. [Conclusions] The addition of ACBM enhances the osteogenic performance of the material, while the plasticity of CPC/CSH contributes to the repair of irregular bone defects. The rapidly fabricated ACBM/CPC/CSH composite artificial bone repair material prepared in this study exhibits excellent biocompatibility and in vivo osteogenic capability. With promising clinical application prospects.