Objective To explore a novel nanometer biomaterial which could induce the regeneration of tooth tissues intell igently, and to evaluate the feasibil ity of using this kind of biomaterial as the scaffold for tooth tissue engineering by investigating the role it plays in tooth tissue engineering. Methods The scaffold for tooth tissue engineering containing recombinant human bone morphogenetic protein 2 (rhBMP-2) was prepared by mixing nanoscale β tricalcium phosphate (β-TCP)/collagen particles. Forty-six 8-10 weeks old specific pathogen free Sprague Dawley (SD)rats, including 34 females and 12 males, weighing 250-300 g, were involved in this study. Tooth germs were removed under a stereomicroscope from the mandible of newborn SD rat, then digested and suspended. Scanning electronic microscope (SEM), adhesion rate of cells, and MTT assay were used to evaluate the effects of the scaffold on the tooth germ cells cultured in vitro. The tissue engineered tooth germ which was constructed by tooth germ cells and scaffold was transplanted under SD rat’s kidney capsule as the experimental group (n=12); the tooth germ cells (cell-control group, n=12) or scaffold without cells (material-control group, n=4) were transplanted separately as control groups Specimens were harvested to perform general and histological observations at 4 and 8 weeks after transplantation. Results β-TCP/collagen showed a loose and porous appearance with soft texture and excellent hydrophil icity. Tooth germ cells grew well and could attach to the scaffold tightly 3 days after coculture. The adhesion rates of tooth germ cells were 27.20% ± 2.37%, 44.52% ± 1.87%, and 73.81% ± 4.15% when cocultured with scaffold for 4, 8, and 12 hours, respectively. MTT assay showed that the cell prol iferation status of experimental group was similar to that of the control group, showing no significant difference (P gt; 0.05). Some white calcified specimens could be harvested at 4-8 weeks after transplantation. At 4 weeks after transplantation some typical structures of dental cusp and enamel-dentin l ike tissues could be seen in the experimental group. Enamel-dentin l ike tissues also formed in some specimens of cell-control group, but they arranged irregularly. At 8 weeks after transplantation the enamel-dentin l ike tissue of experimental group exhibited a mature appearance and organized structure in comparison with that at 4 weeks. And mature enamel or dentin l ike tissue also could be seen in cell-control group. In contrast, there was no enamel or dentin l ike tissue in material-control group at 4 or 8 weeks after transplantation. Conclusion rhBMP-2 decorated β-TCP/collagen scaffold has good biocompatibil ity and can be used as a novel nanometer biomaterial, so it is a good choice in scaffolds for tooth tissue engineering.
【Abstract】 Objective To evaluate the effect on repairing composite defect of mandible and skin by pre fabricatedmusculocutaneous flap including ectopic bone induced by BMP-2 and collagen in rabbits’ latissimus dorsimuscle. Meth ods Twenty-four rabbits (4-6 weeks old) were randomly divided into 3 groups: experimental, control and blank control group (n=8 in each group). Composite carriers composed of BMP-2 and collagen I sponge were implanted into latissimus dorsi muscle pouches of rabbits. The bone formation was evaluated with roentgenography, ALP staining, Von Kossa staining, HE staining, toluidine blue staining and CD31 immunohistochemical labell ing of microvessels. After 6 weeks, the mandibular defect of 8 mm in diameter with local skin defect of 2 cm × 3 cm was made in experimental group, and a musculocutaneous flap including ectopic-induced bone was prefabricated to transfer and repair the composite defect. The mandibular defect of 8 mm in diameter without local skin defect was made in control and blank control group. Free ectopic-induced bone was used for the repair of mandibular defect in control group, but repairing was not performed in blank control group. All the samples were detected 6 weeks after operation for tetracycl ine fluorescent staining, X-ray, histological examination and bone quantity analysis to evaluate the effect. Results Bone formation induced by BMP-2/collagen composites were found as woven bone between 4 to 6 weeks. It showed that cartilaginous osteogenesis was the mainly type of bone formation. Microvessels could beseen in the bony tissues. The composite defects of mandible and skin were healed well in the experimental group. Major bony tissue were seen in the control group, while it still remained bony defect in the blank control group. The bone quantity analysis in the experimental, control, and blank control group were (1.594 ± 0.674), (0.801 ± 0.036), and (0.079 ± 0.010) mm2, there were significant differences between each groups (P lt; 0.05). Conclusion Prefabrication of musculocutaneous flap including boneinduced by the composite of BMP-2 and collagen is feasible and prevalent. It can be regarded as vascularized bone graft and used in repairing composite defect of bone and skin.