Liu Yao, Lin Xiao-ping, Tan Li-si, Wei Wei
Department of Stomatology, the Affiliated Shengqing Hospital of China Medical University, Shenyang 110004, Liaoning Province, China.
Shanghai Kou Qiang Yi Xue. 2006 Dec;15(6):627-31.
Using the MSCs-Bio-Oss tissue engineered bone which was constructed by MSCs as seed cells and the Bio-Oss calf inorganic bone grains as scaffold materials to determine the canine bone formation activity and the feasibility of Bio-Oss combined MSCs to construct tissue engineered bone.
Gybrid canine MSCs were dissociated, cultivated, bone formation induced and differentiated into osteoblast in vitro; The bone formation induced MSCs were allowed to grow onto Bio-Oss calf inorganic bone grains at 10(6) cell/ml, and then incubated and cultivated, under light pressure without other treatments. Inverted phase contrast microscope and scanning electron microscope were used to observe their morphological changes, immunofluorescent labeling of cell surface factor CD44,calcium nodus staining,qualitative and quantitative detection of ALP were carried out, and SPSS 12.0 software package was used for statistical analysis.
The MSCs were uniform with compact alignment and shape of prosenchymatous cells, CD44's surface antigen was positive; Osteoblast activity was present after induction and differentiation, alizarin Bordeaux S stain of calcium nodus was positive, ALP's Gomori staining was also positive. ALP content in the experimental group were (3.307 +/- 0.217) U/g, (5.929 +/- 0.781) U/g and (9.739 +/- 0.547)U/g respectively at 3rd day, 7th day, 14th day after induction and differentiation, which were significantly different (P < 0.01) from the control group: (0.442 +/- 0.087) U/g, (0.581 +/- 0.027)U/g and (0.768 +/- 0.126) U/g; the MSCs stuck compactly on the surface of Bio-Oss, grew well, and formed tissue engineered bone.
Using canine MSCs which were induced by bone formation as seed cells combined with Bio-Oss as scaffold materials to construct tissue engineered bone is feasible. Supported by Liaoning Provincial Natural Science
以间充质干细胞（MSCs）为种子细胞、小牛Bio-Oss无机骨颗粒为支架材料构建MSCs-Bio-Oss组织工程骨，以测定犬骨形成活性及Bio-Oss复合MSCs构建组织工程骨的可行性。
分离、培养杂种犬MSCs，体外诱导其成骨并分化为成骨细胞；将诱导成骨的MSCs以10(6)细胞/ml接种于小牛Bio-Oss无机骨颗粒上，轻压后不做其他处理，进行孵育培养。用倒置相差显微镜、扫描电子显微镜观察其形态变化，进行细胞表面因子CD44免疫荧光标记、钙结节染色、碱性磷酸酶（ALP）定性及定量检测，并采用SPSS 12.0软件包进行统计学分析。
MSCs形态均一，呈梭形细胞紧密排列，CD44表面抗原呈阳性；诱导分化后有成骨细胞活性，钙结节茜素红S染色呈阳性，ALP钙钴法染色也呈阳性。诱导分化后第3天、第7天、第14天实验组ALP含量分别为(3.307±0.217)U/g、(5.929±0.781)U/g、(9.739±0.547)U/g，与对照组(0.442±0.087)U/g、(0.581±0.027)U/g、(0.768±0.126)U/g相比差异有统计学意义（P<0.01）；MSCs紧密黏附于Bio-Oss表面，生长良好，形成组织工程骨。
以成骨诱导的犬MSCs为种子细胞，联合Bio-Oss作为支架材料构建组织工程骨是可行的。辽宁省自然科学基金资助
