Abstract：This study is aimed to conduct fabrication and safety evaluation for the rabbit BMSCs-Cuttlebone composite scaffold. BMSCs were separated by density gradient centrifugation and adherence screening method. The BMSCs-Cuttlebone scaffold was cultivated by precipitation method. The composite scaffold was examined by SEM observation and cell viability assay (MTT test). The biological safety of the composite scaffold was evaluated through muscle implantation test. A total of 40 New Zealand rabbit were randomly divided into two groups, and then BMSCs-cuttlebone scaffold (experimental group) or cuttlebone (control group) were implanted. BMSCs of CD29 positive were obtained by the method of density gradient centrifugation effectively. Scanning electron microscope observation showed that the porosity of cuttlefish bone scaffold was 81.730%±6.770%. The cells adhered on the cuttlebone bracket creeping growth and secreted extracellular matrix. Results obtained from MTT assay showed that the number of viable cells on scaffold increased over time. On the day 1, 3, 5, 7 of cultivation, the cells kept proliferating; while on the day 7 and 8, the proliferation decreased and almost stopped on the day 8. Muscle implantation test showed no significant difference existed between the composite scaffold (experimental group) and cuttlebone group (control group) in the blood routine and blood biochemical tests. In the pathological observation, on the day 7, tissue inflammation grade III-IV for both experimental group and control group; on theday 14, the tissue inflammation grade was II-III for the two groups; on the day28, the tissue inflammation was I-II for the two groups; on the day 56, the tissue inflammation was 0-I grade for the two groups; all of which were within the limit of standard. The rabbit BMSCs could be co-cultured with cuttlebone to build complex biological scaffold. The cells embedded in the scaffold kept proliferation for 7 days. The composite scaffold exhibited good biocompatibility in the muscle implantation test.
