表面矿化修饰煅烧骨/BMP2活性多肽复合MC3T3-E1细胞构建组织工程骨

探讨表面矿化修饰煅烧骨/BMP₂活性多肽复合材料、表面矿化修饰煅烧骨和单纯煅烧骨复合MC3T3-E1细胞构建的组织工程骨在体内的成骨能力。实验分3组，A组：表面矿化修饰煅烧骨/BMP₂活性多肽复合材料，B组：表面矿化修饰煅烧骨，C组：单纯煅烧骨。诱导培养MC3T3-E1细胞，将其分别接种于3组材料，倒置相差显微镜和环境扫描电镜分别观察细胞生长情况。将18只新西兰大白兔随机分成3组，每只大白兔作两侧骶棘肌肌袋模型，然后将3组材料分别植入大白兔骶棘肌内，分别于术后第4、8周各组处死大白兔3只，行组织学观察及新生骨面积测定。通过观察发现MC3T3-E1细胞在3组材料表面生长良好，表面矿化修饰煅烧骨/BMP₂活性多肽复合材料能促进MC3T3-E1细胞在其表面粘附与增殖并能较好地保持细胞的形态。术后第4、8周组织学观察A组材料新生骨明显多于B组，B组多于C组。术后4周和8周新生骨面积测定值A、B、C组分别为（19 712.524±3 782.126）μm²、（28 227.617±2 455.375）μm²，（11 648.507±1 047.221）μm²、（14 592.892±899.532）μm²，（7 986.655±903.487）μm²，（11 254.822±669.508）μm²。表面矿化修饰煅烧骨/BMP₂活性多肽复合材料是一种较理想的骨组织工程复合材料，有望应用于临床。

Tissue Engineered Bone Constructed by Sintered Bone with Surface Mineralization Modification/BMP-2-Related Peptide and MC3T3-E1 Cells

The aim of this study is to investigate the osteogenesis of tissue engineered bone constructed by sintered bone with surface mineralization modification/BMP 2 related peptide (P24) composite with MC3T3-E1 cells in vivo. The biomaterials were divided into three groups: Group A (sintered bone with surface mineralization modification / P24 composite); Group B (sintered bone with surface mineralization modification) and Group C (sintered bone). The MC3T3-E1 cells were induced by osteogenic medium and then seeded into the three materials. The cell material samples were observed by inverted microscopy and environment scanning electron microscopy (ESEM). Eighteen New Zealand white rabbits were divided into three groups with 6 in each group. The model of sacrospinalis myo bag was established by implanting above material samples in the myo bag on the two sides. Every three rabbits in the three groups were killed at the 4^thand 8^th week after implantation, and then the new bone was observed by histologically (HE) and the areas of new osseous tissues were measured. Results from the inverted microscopy and ESEM showed that cells were grown well on the surface of the three materials, and BMP₂ related peptide of Group A was confirmed to improve the adhesion rate and proliferation ability of MC3T3-E1 cells, and maintained the morphology of cells. At different postoperative time points, HE staining results showed new bone of Group A was significantly more than that of Group B and Group C, and that of Group B was significantly more than that of Group C. The areas of the new osseous tissue of group A, B, and C were (19 712.524±3 782.126) μm²,(28 227.617±2 455.375) μm², and (11 648.507±1 047.221) μm² at the 4^thweek post surgery, and (14 592.892±899.532）μm², (7 986.655±903.487)μm², and (11254.822±669.508) μm² at 8^th week post surgery respectively. Sintered bone with surface mineralization modification / P24 composite is an ideal scaffold for bone tissue engineering, which is expected to be applied to clinical studies.