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The osteogenic response of undifferentiated human adipose-derived stem cells under mechanical stimulation

机译:机械刺激下未分化的人类脂肪干细胞的成骨反应

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摘要

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 με, 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.
机译:本研究的目的是研究在机械和/或化学刺激下人脂肪干细胞(hASCs)的成骨反应。 hASC分为三组。在A组中,在没有任何刺激的情况下培养细胞,在B组中,通过化学刺激诱导细胞,而在C组中,通过化学刺激和拉伸负荷的组合诱导细胞。分别使用四点弯曲装置(0.5 Hz,2,000με,2 h / day)和成骨分化培养基施加拉伸负荷和化学刺激。在开始拉伸负荷后的第1、2、3、5和7天,我们通过比色法检测了碱性磷酸酶活性,mRNA表达(RUNX2,ALPL,骨连接蛋白,骨桥蛋白和I型胶原)和蛋白质表达(RUNX2和骨桥蛋白) ,实时PCR和Western blot方法。 B和C组的碱性磷酸酶活性,mRNA表达和蛋白质表达均随培养时间的增加而增加,但在C组的第7天被观察到下调(p <0.05)。与A组相比,B和C组中大多数上述标记均显着较高(p <0.05)。第5天之前,C组所有上述指标均高于B组(p <0.05),但第1天除外。这些结果表明,拉伸负荷促进了hASC的成骨分化,并且相对于单独的化学刺激,机械刺激和化学刺激的组合可以增强至第5天的成骨能力。

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