Safavi Atiyeh Sadat, Rouhi Gholamreza, Haghighipour Nooshin, Bagheri Fatemeh, Eslaminejad Mohamadreza Baghaban, Sayahpour Frough Azam
Faculty of Biomedical Engineering, Amirkabir University of Technology, P. O. Box 1591634311, Tehran, Iran.
National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
In Vitro Cell Dev Biol Anim. 2019 May;55(5):387-394. doi: 10.1007/s11626-019-00340-9. Epub 2019 Apr 15.
This study aimed at investigating the expression of osteoblast and chondrocyte-related genes in mesenchymal stem cells (MSCs), derived from rabbit adipose tissue, under mechanical vibration. The cells were placed securely on a vibrator's platform and subjected to 300 Hz of sinusoidal vibration, with a maximum amplitude of 10 μm, for 45 min per day, and for 14 consequent days, in the absence of biochemical reagents. The negative control group was placed in the conventional culture medium with no mechanical loading. The expression of osteoblast and chondrocyte-related genes was investigated using real-time polymerase chain reaction (real-time PCR). In addition, F-actin fiber structure and alignment with the help of actin filament fluorescence staining were evaluated, and the level of metabolic activity of MSCs was determined by the methyl thiazolyl tetrazolium assay. The real-time PCR study showed a significant increase of bone gene expression in differentiated cells, compared with MSCs (P < 0.05). On the other hand, the level of chondrocyte gene expression was not remarkable. Applying mechanical vibration enhanced F-actin fiber structure and made them aligned in a specific direction. It was also found that during the differentiation process, the metabolic activity of the cells increased (P < 0.05). The results of this work are in agreement with the well-accepted fact that the MSCs, in the absence of growth factors, are sensitive to low-amplitude, high-frequency vibration. Outcomes of this work can be applied in cell therapy and tissue engineering, when regulation of stem cells is required.
本研究旨在探究机械振动作用下,源自兔脂肪组织的间充质干细胞(MSCs)中成骨细胞和软骨细胞相关基因的表达情况。将细胞稳固置于振动器平台上,在无生化试剂的情况下,每天施加300Hz的正弦振动,最大振幅为10μm,持续45分钟,共持续14天。阴性对照组置于无机械负荷的常规培养基中。采用实时聚合酶链反应(实时PCR)研究成骨细胞和软骨细胞相关基因的表达。此外,借助肌动蛋白丝荧光染色评估F-肌动蛋白纤维结构和排列情况,并通过甲基噻唑基四氮唑法测定MSCs的代谢活性水平。实时PCR研究表明,与MSCs相比,分化细胞中骨基因表达显著增加(P<0.05)。另一方面,软骨细胞基因表达水平不显著。施加机械振动增强了F-肌动蛋白纤维结构,并使其沿特定方向排列。研究还发现,在分化过程中,细胞的代谢活性增加(P<0.05)。本研究结果与已被广泛接受的事实一致,即在无生长因子的情况下,MSCs对低振幅、高频振动敏感。当需要调控干细胞时,本研究结果可应用于细胞治疗和组织工程。