| ObjectiveTo reconstruct Mandibular defect using tissue engineering bone with BMSCs cell sheets and investigate the effect of cell sheets on osteogenesis.MethodsBone marrow stem cells were isolated with the method of density gradient centrifugation from canine and cultured. BMSCs were induced to differentiate to osteoblasts. The BMSCs induced were transferred to the temperature-responsive culture dishes and cultured. BMSCs cell sheets were formed by temperature changed. VEGF, rhBMP-2 and cell suspension were added in the PLGA scaffolds. The PLGA scaffolds wrapped with cell sheets were implanted into the Mandibular defect in the right side (experimental side). The PLGA scaffolds without cell sheets were implanted into the left side of mandibular as control side.16 dogs were evenly divided into 4 groups, and each group were executed after every 4,8,12,16 weeks for the investigation of gross, histological and radiological observation and hardness test.ResultsThe Osteogenesis of experimental side was better than that of control side.16 weeks after implantation, most area of the Mandibular defect was replaced by fresh bone tissues. Compact bone similar with normal bone tissue was formed in the lingual defect of mandible and had bony union with the bone stump. The optical density of the fresh bone in the experimental side was higher than that of the control side, there was a significant difference between the two sides (p<0.05). Plenty of lamellar bones were formed in experimental side and Haversian system and red marrow were observed. After 8 weeks, the hardness of the specimens rise as time increase and the experimental side was higher than that of the control side, there was a significant difference between the two sides (p<0.05).ConclusionTissue engineering bone with the structure of lamellar bones can be formed by the technology of BMSCs cell sheets. |
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