Restoration of Bone Defects Using Modified Heterogeneous Deproteinized Bone Seeded with Bone Marrow Mesenchymal Stem Cells

The aim of the present study was to investigate the effect of modified heterogeneous deproteinized bone combined with bone marrow mesenchymal stem cells (BMSCs) in the restoration of a validated bone defect model. BMSCs were identified by flow cytometry and multilineage differentiation assay. The structural features of the modified heterogeneous deproteinized bone scaffold and biocompatibility between BMSCs and the scaffold were confirmed by scanning electron microscope (SEM) detection. The cytotoxicity of the modified heterogeneous deproteinized bone scaffolds were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenytetrazolium bromide (MTT) assay. SEM detection proved that modified heterogeneous deproteinized bone scaffold had no negative impact on the proliferation of BMSCs. MTT assay results demonstrated that the scaffold had no apparent cytotoxicity. Biomechanical detection showed that the stiffness and ultimate loading of tibias in the scaffold + BMSCs group were significantly higher than those of the scaffold alone group (P < 0.05) and the control group (P < 0.01). Histological analyses confirmed that the greatest quantity of new bone was generated in the scaffold + BMSCs group, when compared with all other groups, at 8 weeks' post-operation. The bone mineral density (BMD) in the scaffold + BMSC group was significantly higher than that of the scaffold alone group (P < 0.05) and the control group (P < 0.01). Fluorometric analyses confirmed the presence of BMSCs at high concentration within the bone defect areas in the scaffold + BMSCs group at 4 weeks after transplantation. These findings suggest that the modified heterogeneous deproteinized bone scaffold seeded with BMSCs can effectively enhance the restoration of bone defects.