流体剪切力作用下人脂肪基质细胞体外成骨分化的实验研究

口腔医学研究 ›› 2015, Vol. 31 ›› Issue (10): 1020-1023.

流体剪切力作用下人脂肪基质细胞体外成骨分化的实验研究

崔军^*,贾黎,刘学恒,崔光辉

北京大学深圳医院口腔科广东深圳 518000

收稿日期:2015-01-26 出版日期:2015-10-28 发布日期:2016-05-04
通讯作者: 崔军,电话:0755-83923333-8446
作者简介:崔军(1979~ ),男,陕西人,副主任医师,博士,主要从事口腔修复学及口腔种植学临床和基础的研究工作。
基金资助:
深圳市知识创新计划基础研究项目(编号:20130320180202)

Study of Fluid Shear Stress on Osteogenic Differentiation of Human Adipose-derived Stromal Cells in Vitro

CUI Jun, JIA Li, LIU Xue-hen, CUI Guang-hui

Department of Stomatology, Peking University Shenzhen Hospital, Shenzhen 518000, China

Received:2015-01-26 Online:2015-10-28 Published:2016-05-04

摘要/Abstract

摘要： 目的:分离人体脂肪基质细胞(hADSCs)并研究流体剪切应力对其成骨分化的影响。方法:将脂肪抽吸术获取的人体脂肪进行分离,应用强度为3 dyne/cm²的流体剪切力刺激30 min后观察细胞形态;应用四甲基偶氮唑蓝(MTT)法检测细胞的增殖能力, 绘制细胞生长曲线;碱性磷酸酶(ALP)及茜素红染色对其成骨分化进行鉴定。结果:胶原酶消化后的脂肪基质细胞呈梭形, 经流体剪切力作用培养后,细胞体积明显增大,胞浆丰富并含有较多的细胞器,细胞排列方向与加力方向一致, 呈漩涡状生长,生长速度加快。此外,流体剪切力加载后脂肪基质细胞内碱性磷酸酶活性增高,茜素红染色表明聚集的细胞团中心能形成矿化结节。结论:脂肪基质细胞中的成体干细胞经过流体剪切力作用后可向成骨细胞分化并具有明显的成骨表型,可作为骨组织工程的种子细胞。

关键词: 流体剪切力, 脂肪基质细胞, 体外骨分化, 骨组织工程

Abstract: Objective: To investigate the influence of fluid shear stress (FSS) on osteogenic differentiation of human adipose-derived stromal cells (hADSCs) in vitro. Methods: hADSCs were cultured and proliferated in vitro, and subjected to a FSS of 3 dyne/cm2 for 30min every day. Then the hADSCs were observed under inverted microscope. The growth curve was evaluated by cell differentiation ability which was measured by MTT. Moreover, the osteoblastic differentiation ability was also tested by ALP activity and mineralization activity. Results: After treated by FSS, hADSCs were characterized by large size with a relatively large nucleus and plenty cytoplasm. The orientation of hADSCs growth was consistent with the direction of FSS and the growth rate was enhanced by FSS. What's more, ALP activity and mineralization activity of hADSCs were both increased significantly after FSS stimulation. Conclusion: hADSCs exhibited the osteoblastic differentiation ability after treated with FSS, which could be used as the seed cells for bone tissue engineering.

Key words: Fluid shear stress, Adipose-derived stromal cell, Osteogenic differentiation, Bone tissue engineering

中图分类号:

R782.2

崔军,贾黎,刘学恒,崔光辉. 流体剪切力作用下人脂肪基质细胞体外成骨分化的实验研究[J]. 口腔医学研究, 2015, 31(10): 1020-1023.

CUI Jun, JIA Li, LIU Xue-hen, CUI Guang-hui. Study of Fluid Shear Stress on Osteogenic Differentiation of Human Adipose-derived Stromal Cells in Vitro[J]. Journal of Oral Science Research, 2015, 31(10): 1020-1023.

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