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目的:观察血管内皮祖细胞(endothelial progenitor cells,EPCs)在实验犬体内构建组织工程骨时对其成骨能力的影响。方法:抽取实验犬骨髓,密度梯度离心法获取单个核细胞,通过不同培养方法获得经成骨诱导的骨髓间充质干细胞(bone marrow stromal cells,BMSCs)和EPCs,在体外复合犬同种异体脱钙骨基质(demineralized bone matrix,DBM),扫描电镜和相差显微镜观察细胞形态和生长情况;将复合体植入实验犬背阔肌筋膜下,右侧为实验组,植入EPCs/BMSCs/DBM复合体,左侧为对照组,植入BMSCs/DBM复合体。影像学和组织学方法观察术后不同时间点组织工程骨的骨形成程度。采用SPSS13.0软件包对所得数据进行t检验。结果:术后4、8、12周,X线显示实验组的骨密度显著高于对照组,差异有统计学意义(P<0.05);组织学观察显示,实验组成骨多于对照组,2组新生骨面积及血管面积之间差异有统计学意义(P<0.05)。结论:EPCs可以促进组织工程骨的成骨能力,加速新生骨形成。
OBJECTIVE: To observe the effect of endothelial progenitor cells (EPCs) on the osteogenic ability of tissue engineered bone in experimental dogs. Methods: Mononuclear cells were isolated from the canine bone marrow by density gradient centrifugation. Bone marrow-derived bone marrow stromal cells (BMSCs) and EPCs were obtained by different culture methods. (DBM), scanning electron microscopy (SEM) and phase contrast microscopy. The composite was implanted into the fascia of the latissimus dorsi and the right side was the experimental group. EPCs / BMSCs / DBM Complex, the left side of the control group, implanted BMSCs / DBM complex. Imaging and histological methods were used to observe the degree of bone formation of tissue engineered bone at different time points after operation. SPSS 13.0 software package was used to test the data t test. Results: At 4, 8, and 12 weeks after operation, the X-ray showed that the BMD of the experimental group was significantly higher than that of the control group (P <0.05). Histological observation showed that the experimental group had more bone than the control group There was significant difference between the newborn bone area and the vascular area (P <0.05). Conclusion: EPCs can promote the osteogenic capacity of tissue engineered bone and accelerate the formation of new bone.