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作 者:王玲玲[1] 李宁毅[1] 樊功为[1] 卜令学[1] 杨学财[1] 高振华[1]
机构地区:[1]青岛大学医学院附属医院口腔颌面外科,山东青岛266003
出 处:《中国口腔颌面外科杂志》2011年第2期96-100,共5页China Journal of Oral and Maxillofacial Surgery
基 金:国家自然科学基金(30872896);山东省自然科学基金(Y2008C77)
摘 要:目的:探讨犬骨髓基质细胞(bone marrow stromal cells,BMSCs)细胞片层在构建组织工程骨中的价值。方法: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(rhBMP-2)复合到DBM上。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)。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,植入犬背阔肌血运丰富的肌筋膜下为实验侧,以无BMSCs细胞片层包裹的DBM/rhBMP-2/BMSCs复合体为对照侧。术后4、8、12周取材,行组织学观察,评价体内异位成骨的情况。采用SPSS13.0软件包对数据进行两样本均数差别的t检验。结果:实验侧成骨面积大于对照侧,2组差异有显著性(P<0.05)。术后12周,实验侧生成大量板层骨,有哈弗系统形成,骨髓腔内有红骨髓。对照侧有板层骨形成,无哈弗系统形成,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系统的组织工程骨的形成。PURPOSE: To investigate the effect of bone marrow stromal cell sheet on the formation of tissue-engineered bone in dogs.METHODS: Demineralized bone matrix(DBM) were prepared from homologous bone.DBM was constituted with recombination human bone morphogenetic protein-2(rhBMP-2).And bone marrow stromal cells(BMSCs) were isolated from iliac bone of dogs with the method of density gradient centrifugation in vitro.BMSCs induced by osteogenic DMEM at passage 3 were incubated in the temperature-responsive culture dish to form BMSCs cell sheet.BMSCs cell sheet combined with DBM/rhBMP-2/BMSCs was implanted around the vessels of latissimus dorsi muscle in the experimental side,and DBM/rhBMP-2/BMSCs without BMSCs cell sheet was implanted around the vessels of latissimus dorsi muscle in the control side.4,8,12 weeks after operation,the ectopic bone formation was investigated by histological observation.The Data were analyzed by two independent samples t-test with SPASS 13.0 software package.RESULTS: Osteogenesis result was experimental sidecontrol side.Newly formed bone tissue of two sides was significantly different(P0.05).12 weeks after operation,a large number of lamellar bone and Haversian system formed in the experimental side,with red bone marrow in the bone marrow cavity.Lamellar bone formed in the control side without Haversian system and red bone marrow.CONCLUSION: BMSCs cell sheet could promote the formation of tissue-engineered bone with dense lamellar bone and Haversian system.Supported by National Natural Science Foundation of China(30872896) and Natural Science Foundation of Shandong Province(Y2008C77).
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