摘要
目的探讨犬骨髓基质细胞(bone marrow stromal cells,BMSCs)片层在构建组织工程骨中的价值。方法抽取犬髂骨骨髓,采用密度梯度离心法分离犬骨髓基质细胞(BMSCs)。制备犬同种异体脱钙骨基质(dermineralized bone matrix,DBM)。将人重组骨形态发生蛋白-2(recombination human bone morphogenetic protein-2,rhBMP-2)复合到DBM上。将经成骨诱导的第3代细胞接种于温度反应性培养皿中,制备BMSCs细胞片层。用得到的BMSCs细胞片层包裹DBM/rhBMP-2/BMSCs复合体,植入犬左侧背阔肌肌筋膜下为实验组,以无BMSCs细胞片层包裹的DBM/rhBMP-2/BMSCs复合体植入犬右侧背阔肌肌筋膜下为对照组。术后8、12、16周取材行组织学观察,评价体内异位成骨的情况。结果成骨面积实验组(对照组,两组差异有显著性(P<0.05)。术后16周,实验组有大量板层骨形成,有哈弗氏系统形成,骨髓腔内有红骨髓。对照组有板层骨形成,无哈弗氏系统形成,骨髓腔内无红骨髓。结论 BMSCs细胞片层可促进具有致密板层骨和哈弗氏系统的组织工程骨的形成。
Objective To investigate the effect of bone marrow stromal cell sheet on the construction of tissue-engineered bone in dogs.Methods Bone marrow stromal cells(BMSCs) were isolated from iliac bone of dogs with the method of density gradient centrifugation in vitro.Demineralized bone matrix(DBM) were prepared from homologuous bone.DBM was composited with recombination human bone morphogenetic protein-2(rhBMP-2).BMSCs induced by osteogenic DMEM at passage 3 were incubated in the temperature-responsive culture dish to make the BMSCs cell sheet.BMSCs cell sheet combined with DBM/rhBMP-2/BMSCs was implanted around the vessels of lattisimus dorsi muscle on the left side in the experiment group,and DBM/rhBMP-2/BMSCs without BMSCs cell sheet was implanted around the vessels of lattisimus dorsi muscle on the right side in the control group.8,12,16 weeks after operation,the bone formation was investigated by histological observation.Results Osteogenesis result was experimental group>control group.Newly formed bone tissue in two groups were significantly different(P<0.05).16 weeks after operation,a large number of lamellar bone and haversian system formed in experimental group,with red bone marrow in the bone marrow cavity.Lamellar bone formed in control group without haversian system and red bone marrow.Conclusions BMSCs cell sheet could promote the formation of tissue-engineered bone with dense lamellar bone and haversian system.
出处
《口腔医学》
CAS
2011年第4期199-202,209,共5页
Stomatology
基金
国家自然科学基金资助项目(30872896)
山东省自然科学基金资助项目(Y2008C77)
关键词
骨髓基质细胞
细胞片层
温度反应性培养皿
组织工程骨
犬
bone marrow stromal cells
cell sheet
temperature responsive culture dish
tissue-engineered bone
dog
