骨髓基质干细胞片层的制备及其在组织工程骨中应用的初步研究

目的：探讨骨髓基质干细胞（bone marrow stromal cells,BMSCs）片层（cell sheet）的制备方法,并观察其在实验犬体内构建组织工程骨时的成骨作用。方法：密度梯度离心法获取实验犬骨髓基质干细胞,经成骨诱导培养后,利用温度感应性培养皿（temperature-responsive culture dish）制备细胞片层后在体外包裹犬同种异体脱钙骨基质（demineralized bone matrix,DBM）,相差显微镜和扫描电镜观察细胞及片层形态;将复合体植入实验犬背阔肌筋膜下,组织学方法观察术后组织工程骨的骨形成情况。结果：细胞片层成功制备,并与支架材料复合良好,术后16周组织学显示形成类似板层骨的致密骨组织。结论：利用温度反应性培养皿可成功制备BMSC细胞片层,细胞片层包裹支架材料后在实验动物体内可形成具有类似板层骨结构的组织工程骨。

A preliminary study on fabrication of BMSCs cell sheet and its application in bone tissue engineering

Objective：This study was designed to investigate the fabrication of bone marrow stromal cells （BMSCs） sheet and its application in bone tissue engineering.Methods：BMSCs were derived from dog bone marrow and cell sheets were prepared with temperature-responsive dishes after cells were osteogenic induced.The sheets were wrapped on demineralized bone matrix （DBM）,BMSCs and cell sheet were observed under inverted microscope and scanning electron microscope,then the construct was implanted into the bottom fasciae of latissimus dorsi muscle,bone formation was observed with histological methods after operation.Results：BMSCs sheet was successful prepared and new bone formation with high density was observed 16 weeks after operation.Conclusion：BMSCs sheet can be fabricated with temperature-responsive culture dishes and it can promote new bone formation with similar lamellar structure in bone tissue engineering.