犬同种异体脱钙骨基质复合细胞片层的体外培养与观察(英文)

Complex of dog allogenic decalcified bone matrix and bone marrow stromal cell sheets In vitro culture and observation

背景:如何构建具有类似天然骨结构组织工程骨的问题是组织工程学发展的一个难题。细胞片层技术的出现,为其提供了新的途径。目的:观察细胞片层与脱钙骨基质的生物相容性及其在脱钙骨基质上的生长情况。设计、时间及地点:体外观察性实验,于2009-06/2009-09在青岛大学医学院附属医院中心实验室完成。材料:利用温度感应培养基制备犬骨髓基质细胞片层;脱脂、脱钙、脱非胶原蛋白方法制备犬脱钙骨基质。方法:将温度感应技术制备刮取的细胞片层铺盖于脱钙骨基质表面,用含体积分数为10%胎牛血清及成骨诱导剂的DMEM培养液浸没培养。主要观察指标:扫描电镜下观察脱钙骨基质的结构及细胞片层在脱钙骨基质上的附着、生长情况。计算脱钙骨基质孔隙率和孔径大小。结果:扫描电镜下观察可见脱钙骨基质呈三维立体网孔结构。材料孔隙率约为75%。平均孔隙直径为(250.11±98.89)μm,成正态性分布。扫描电镜下观察细胞片层在脱钙骨基质上的附着、生长状况良好,增殖迅速。结论:细胞片层与脱钙骨基质有较好的生物相容性,脱钙骨基质/细胞片层复合体能够应用于组织工程骨,为构建类似骨结构组织工程骨起到促进作用。

BACKGROUND：How to reconstruct tissue-engineered bone with structure similar to natural bone is a problem in the development of tissue engineering.Cell sheet engineering technology enables novel approaches to construction of tissue-engineered bone.OBJECTIVE：To observe the biocompatibility of cell sheets to decalcified bone matrix（DBM）and their growth on DBM.DESIGN,TIME AND SETTING：An in vitro observation was performed at the Central Laboratory,Affiliated Hospital,Qingdao University Medical College between June and September 2009.MATERIALS：Dog bone marrow stromal cell sheets were prepared using temperature-responsive medium.Dog DBM was prepared by defatting,decalcification,and noncollagen protein removal procedures.METHODS：DBM surface was covered by cell sheets prepared by temperature-responsive technology and cultured with DMEM containing 10%fetal bovine serum and osteoinductive agent.MAIN OUTCOME MEASURES：Under scanning electron microscope,DBM structure,as well as the attachment and growth of cell sheets on DBM surface,was observed.Porosity and aperture size of DBM were calculated.RESULTS：DBM exhibited a three-dimensional latticed structure,with a porosity of approximately 75%.The mean aperture size was（250.11±98.89）μm,exhibiting a normal distribution.Cell sheets well attached to and grew on DBM surface,and rapidly proliferated.CONCLUSION：Cell sheets show good biocompatibility to DBM.DBM/cell sheets complex can be applied in tissue-engineered bones,which promotes the construction of tissue-engineered bone with structure similar to natural bone.