Du Chun-hua, Li Ning-yi, Gao Ning, Yao Chao, Wang Shuang-yi, Bu Ling-xue
Central Laboratory, Affiliated Hospital of the Medical College, Qingdao University, Qingdao, China.
J Oral Maxillofac Surg. 2013 Sep;71(9):1531.e1-1531.e10. doi: 10.1016/j.joms.2013.04.029. Epub 2013 Jul 15.
The aim of this study was to construct functional tissue-engineered bone in dogs using cell sheet engineering, a new technique to gain and transfer seed cells.
Demineralized bone matrixes, prepared from homologous bone, were coated with recombinant human bone morphogenetic protein-2. Bone marrow stromal cells (BMSCs) were isolated and subcultured. Osteogenic-induced BMSCs were incubated in a temperature-responsive culture dish to form the BMSC sheet. The complex of demineralized bone matrix, recombinant human bone morphogenetic protein-2, and BMSCs wrapped with BMSC sheets was implanted around the blood vessels of the latissimus dorsi muscle in the experimental side, and the same complex without BMSC sheets was implanted around the blood vessels of the latissimus dorsi muscle on the other side as a control. At 4, 8, 12, and 16 weeks after implantation, the implants were removed for radiographic evaluation, descriptive histologic observation, and histologic quantitative analysis.
Radiographic analysis showed that the optical density of the tissue-engineered bone on the 2 sides increased with time. However, the optical density of the experimental side was significantly greater than that of the control side at the same points. Sixteen weeks after implantation, mature lamellar bone was formed in the experimental side, with red bone marrow in the bone marrow cavity. In contrast, the control side exhibited significantly less lamellar bone. Histologic quantitative analysis showed that the experimental side exhibited significantly more bone per area compared with the control side.
BMSC sheet engineering may be useful to construct functional tissue-engineered bone.
本研究的目的是利用细胞片工程技术在犬体内构建功能性组织工程骨，这是一种获取和转移种子细胞的新技术。
用重组人骨形态发生蛋白-2包被由同种异体骨制备的脱矿骨基质。分离并传代培养骨髓基质细胞（BMSCs）。将经成骨诱导的BMSCs接种于温度响应培养皿中形成BMSC片。将脱矿骨基质、重组人骨形态发生蛋白-2与包裹有BMSC片的BMSCs复合物植入实验侧背阔肌血管周围，另一侧背阔肌血管周围植入不含BMSC片的相同复合物作为对照。植入后4、8、12和16周，取出植入物进行影像学评估、描述性组织学观察和组织学定量分析。
影像学分析显示，两侧组织工程骨的光密度均随时间增加。然而，在相同时间点，实验侧的光密度明显高于对照侧。植入16周后，实验侧形成成熟的板层骨，骨髓腔内有红骨髓。相比之下，对照侧板层骨明显较少。组织学定量分析显示，与对照侧相比，实验侧每单位面积的骨量明显更多。
BMSC片工程技术可能有助于构建功能性组织工程骨。
