| Objective:To investigate the effect of bone marrow stromal cell sheet on the construction of tissue-engineered bone in dogs.Methods:Demineralized bone matrix(DBM) were prepared from homologuous bone. DBM were composited with recombination human bone morphogenetic protein-2(rhBMP-2). Bone marrow stromal cells(BMSCs) were isolated from iliac bone of dogs with the method of density gradient centrifugation in vitro. BMSCs were induced to differentiate to osteoblasts by osteogenic DMEM. BMSCs induced at passage 3 were incubated in the temperature-responsive culture dish, when BMSCs fused with each other, the temperature of the incubator was changed from 37℃to 20℃to form the BMSCs cell sheet. BMSCs cell sheet combined with DBM/rhBMP-2/BMSCs wrap the thoracodorsal artery and vein of the dog was implanted at the left side in experimental side, and DBM/rhBMP-2/BMSCs without BMSCs cell sheet wrap the thoracodorsal artery and vein of the dog was implanted at the right side in control side.8,12,16 weeks after operation, the bone formation was investigated by gross observation, imaging obserbation and histological observation. The Data were analyzed by two independent samples t-test with SPASS 18.0 software package.Results:1. The prepared BMSCs cell sheet was complete and compact, and grew well on the surface of the scaffold complex.2. Osteogenesis result was experimental side>control side. Newly formed bone tissue of two sides was significantly different(P<0.05).16 weeks after operation, a large number of lamellar bone and haversian system formed with red bone marrow in bone marrow cavity in experimental side. Lamellar bone formed without haversian system and red bone marrow in control side.Conclusion:BMSCs cell sheet could promote the formation of tissue-engineered bone with dense lamellar bone and haversian system. |
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