犬脂肪基质干细胞复合珊瑚支架材料裸鼠皮下成骨的实验研究  

Experimental Study of Subcutaneous Osteogenesis by Adscs with Coral Scaffolds

作　　者：沈海刚[1] 李晔[1] 刘波[2] 袁捷[3] 

机构地区：[1]杭州市中医院外一科,浙江杭州310007 [2]浙江省中医院,浙江杭州310006 [3]上海市第九人民医院,上海200011

出　　处：《中华中医药学刊》2012年第7期1548-1550,I0005,I0006,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：浙江省自然科学基金资助项目(Y2080180);杭州市卫生局重点科技项目(2003Z006)

摘　　要：目的:探讨犬脂肪基质干细胞(adipose derived stromal cells,ADSCs)作为种子细胞复合珊瑚支架材料在裸鼠皮下成骨的可行性。方法:体外分离培养、成骨诱导、扩增犬ADSCs,将第2代细胞接种于珊瑚支架上,形成细胞-材料复合物。倒置相差显微镜观测细胞形态,DIO染色后激光共聚焦荧光显微镜观察细胞在材料上的生长状况,细胞材料复合物植于8只裸鼠右侧背部皮下作为实验组,在左侧对称部位植入单纯珊瑚支架作为对照组。分别在移植后第4、8周取材,HE染色,进行组织学及组织形态计量学分析。结果:ADSCs倒置相差显微镜下为成纤维细胞样梭形外观,与材料共培养后共聚焦显微镜可见细胞贴附于材料表面及孔隙内生长。HE染色显示,4周时实验组可见新生骨组织形成;8周时实验组新生骨组织形成增加。图像分析显示,两个时间点实验组成骨量与对照组相比差异皆有统计学意义(P<0.05);实验组第8周成骨量高于第4周,差异也具有统计学意义(P<0.05)。结论:犬ADSCs作为种子细胞复合珊瑚支架材料可以在裸鼠皮下形成骨样组织。Objective：This study was to investigate osteogenesis of canine ADSCs complex with coral scaffolds implanted subcutaneously.Methods：ADSCs were isolated and cultured in osteogenic-induced medium.The induced cells were complex with coral to co-culture.The ADSCs were observed under inverted microscope.The constructs were examined from confocal microscopy.The constructs were implanted into the subcutaneous tissue of the nude mice as experimental group,and the coral as control group.The engineered tissue was harvested in 4 and 8 weeks post-implantation and processed for HE staining,then evaluated by histology and histomorphometry.Results：The ADSCs appeared fibroblast-like under inverted microscope.Confocal microscopy showed ADSCs grew well on the coral.HE staining showed a small group of woven bone formation 4 weeks later in the experimental group,and more woven bone formation were observed 8 weeks later in the experimental group.Histomorphometry showed more woven bone formation in the experimental group than the control group at 4 and 8 weeks later（P〈0.05）.Conclusions：The ADSCs/coral composites implanted subcutaneously in nude mice could construct woven bone.

关 键 词：脂肪基质干细胞 珊瑚 裸鼠 

分 类 号：R-33[医药卫生]