The Osteogenic Response of Undifferentiated Human Adipose-Derived Stem Cells under Mechanical Stimulation
The Osteogenic Response of Undifferentiated Human Adipose-Derived Stem Cells under Mechanical Stimulation
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DOI：
10.1159/000335905
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发表时间：
2012-01-01
影响因子：
2.7
通讯作者：
Long, Jie
中科院分区：
文献类型：
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作者：
Du, Hong-ming;Zheng, Xiao-hui;Long, Jie
本研究的目的是研究人类脂肪源性干细胞（hASCs）在机械和/或化学刺激下的成骨反应。hASCs分为三组。A组细胞不进行任何刺激培养，B组细胞采用化学刺激诱导，C组细胞采用化学刺激和拉伸载荷联合诱导。拉伸载荷和化学刺激分别采用四点弯曲仪（0.5 Hz, 2000 μ epsilon, 2 h/day）和成骨分化培养基。在拉伸加载开始后的第1、2、3、5和7天，分别采用比色法、实时荧光定量PCR和Western blot法检测小鼠碱性磷酸酶活性、mRNA表达（RUNX2、ALPL、骨连接素、骨桥蛋白和I型胶原）和蛋白表达（RUNX2和骨桥蛋白）。B组和C组碱性磷酸酶活性、mRNA表达量和蛋白表达量均随培养时间的延长而升高，而C组碱性磷酸酶活性、mRNA表达量和蛋白表达量在培养第7天呈下降趋势（p < 0.05）。与A组相比，B、C组上述指标均显著升高（p < 0.05）。除第1天外，第5天前C组以上各项指标均高于B组（p < 0.05）。这些结果表明，拉伸载荷促进了hASCs的成骨分化，机械和化学联合刺激相对于单独的化学刺激可以提高hASCs的成骨能力至第5天。版权所有(c) 2012 S. Karger AG，巴塞尔
The purpose of this study was to investigate the osteogenic response of human adipose-derived stem cells (hASCs) under mechanical and/or chemical stimulation. hASCs were divided into three groups. In group A, the cells were cultured without any stimulation, in group B, the cells were induced with chemical stimulation, and in group C, the cells were induced with a combination of chemical stimulation and stretch loading. Stretch loading and chemical stimulation were applied using a four-point bending apparatus (0.5 Hz, 2,000 mu epsilon, 2 h/day) and osteogenic differentiation medium, respectively. At the 1st, 2nd, 3rd, 5th and 7th day following initiation of stretch loading, we detected alkaline phosphatase activity, mRNA expression (RUNX2, ALPL, osteonectin, osteopontin and type I collagen) and protein expression (RUNX2 and osteopontin) by colorimetric assay, real-time PCR and Western blot methods, respectively. Alkaline phosphatase activity, mRNA expression and protein expression all increased in groups B and C along with the culture time, but were observed to be downregulated by the 7th day in group C (p < 0.05). Compared to group A, most of the above markers were significantly higher in groups B and C (p < 0.05). All of the above markers in group C were higher than those in group B before the 5th day (p < 0.05), except at the 1st day. These results indicated that stretch loading promoted osteogenic differentiation of hASCs and that the combination of mechanical and chemical stimulation could enhance the osteogenic capability up to the 5th day relative to chemical stimulation alone. Copyright (c) 2012 S. Karger AG, Basel