Effects of cyclic mechanical stretching on the mRNA expression of tendon/ligament-related and osteoblast-specific genes in human mesenchymal stem cells
Effects of cyclic mechanical stretching on the mRNA expression of tendon/ligament-related and osteoblast-specific genes in human mesenchymal stem cells
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DOI：
10.1080/03008200701818561
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发表时间：
2008-01-01
影响因子：
2.9
通讯作者：
Young, Tai-Horng
中科院分区：
文献类型：
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作者：
Chen, Yi-Jane;Huang, Chien-Hsun;Young, Tai-Horng
本研究的目的是探讨周期性机械拉伸对肌腱/韧带相关和成骨细胞特异性标记基因的mRNA表达的影响，在人骨髓间充质干细胞接种到I型胶原涂层表面。通过定量实时PCR分析拉伸诱导的间充质干细胞蛋白（MSCP）、基质金属蛋白酶-3（MMP-3）、肌腱/韧带细胞相关标志基因（I型胶原、III型胶原和腱生蛋白-C）以及成骨细胞典型标志基因（核心结合因子α 1（Cbfa 1）、碱性磷酸酶（ALP）和骨钙素（OCN））的mRNA表达。结果显示，无论拉伸的大小（高或低），在受到机械负荷的MSC中，MSCP的显著下调和MMP-3基因的上调。此外，成骨细胞谱系的典型标记基因被低幅度拉伸上调，而肌腱/韧带相关基因被长时间高幅度拉伸上调。Cbfa 1和ALP早在8小时开始上调，随后呈下降趋势，在其他时间点表达无显著变化。10%拉伸48 h后，I型胶原、III型胶原和腱生蛋白C的mRNA表达显著增加，拉伸48 h后，这种效应仍然存在。本研究证实了周期性机械拉伸对不同细胞系相关标志基因差异转录的影响。低强度牵张可增加Cbfa 1和ALP的mRNA表达，并可能参与MSCs的早期成骨分化，而高强度牵张可上调肌腱/韧带相关基因的mRNA表达。
The purpose of this study was to explore the influences of cyclic mechanical stretching on the mRNA expressions of tendon/ligament-related and osteoblast-specific marker genes in human MSCs seeded onto a collagen type I-coated surface. The stretch-induced mRNA expressions of mesenchymal stem cell protein (MSCP), matrix metalloproteinase-3 (MMP-3), and marker genes related to tendon/ligament cells (type I collagen, type III collagen, and tenascin-C) and those typical of osteoblasts (core binding factor alpha 1 (Cbfa1), alkaline phosphatase (ALP), and osteocalcin (OCN)) were analyzed by quantitative real-time PCR. The results revealed significant downregulation of MSCP and upregulation of MMP-3 genes in MSCs subjected to mechanical loading, regardless of the magnitude of the stretching (high or low). Moreover, the typical marker genes of the osteoblast lineage were upregulated by low-magnitude stretching, whereas tendon/ligament-related genes were upregulated by high-magnitude stretching for a long period. Cbfa1 and ALP were upregulated starting as early at 8 hr, followed by a downward trend and no significant change in expression at the other time points. The mRNA expressions of type I collagen, type III collagen, and tenascin-C significantly increased in MSCs subjected to 10% stretching for 48 hr, and this effect still existed after the stretched cells had rested for 48 hr. This study demonstrated the effect of cyclic mechanical stretching on differential transcription of marker genes related to different cell lineages. Low-magnitude stretching increased mRNA expressions of Cbfa1 and ALP and was possibly involved in the early osteoblastic differentiation of MSCs, whereas high-magnitude stretching upregulated the mRNA expressions of tendon/ligament-related genes.