Department of Medical Biotechnology, Dongguk University, Seoul, Republic of Korea.
Connect Tissue Res. 2012;53(2):149-59. doi: 10.3109/03008207.2011.619284. Epub 2011 Dec 7.
In this study, we evaluated the effect of mechanical stimulation on the differentiation of umbilical cord-derived mesenchymal stem cells (UC-MSCs) in osteogenic medium using a Flexcell system that imposed cyclic uniaxial mechanical stimulation at a strain of 0%, 5%, or 10% (5 s of stretch and 15 s of relaxation) for 10 days. The expression of MSC surface antigens (CD73, CD90, and CD105) was significantly decreased as strain increased. Mechanical stimulation inhibited the growth of UC-MSCs and slightly raised lactate dehydrogenase production. Mechanically stimulated groups produced more elastin and sulfated glycosaminoglycan than unstimulated groups and these increases were in proportion to the degree of strain. Reverse transcription-polymerase chain reaction analysis revealed that mechanical stimulation induced a significant increase in the mRNA expression of osteoblast differentiation markers. The mRNA levels of osteopontin, osteonectin, and type I collagen in the 5% and 10% strained groups were significantly higher than those in the 0% strained group. From the Western blot analysis, UC-MSCs produced bone sialoprotein and vimentin in a mechanical strain-dependent manner. Thus, cyclic mechanical loading was able to enhance the differentiation of human UC-MSCs into osteoblast-like cells as determined by osteogenic gene and protein expression. Furthermore, this finding has important implications for the use of the combination of mechanical and osteogenic differentiation media for UC-MSCs in tissue engineering and regenerative medicine.
在这项研究中,我们使用 Flexcell 系统评估了在成骨培养基中机械刺激对脐带间充质干细胞(UC-MSCs)分化的影响,该系统以 0%、5%或 10%的应变量(拉伸 5 秒和松弛 15 秒)施加循环单轴机械刺激 10 天。随着应变的增加,MSC 表面抗原(CD73、CD90 和 CD105)的表达显著降低。机械刺激抑制 UC-MSCs 的生长,略微提高乳酸脱氢酶的产生。与未刺激组相比,机械刺激组产生更多的弹性蛋白和硫酸化糖胺聚糖,并且这些增加与应变程度成比例。逆转录-聚合酶链反应分析显示,机械刺激诱导成骨细胞分化标志物的 mRNA 表达显著增加。5%和 10%应变组的骨桥蛋白、骨连接素和 I 型胶原的 mRNA 水平明显高于 0%应变组。从 Western blot 分析可知,UC-MSCs 以机械应变依赖性的方式产生骨涎蛋白和波形蛋白。因此,循环机械加载能够增强人 UC-MSCs 向成骨样细胞的分化,这可通过成骨基因和蛋白表达来确定。此外,这一发现对于在组织工程和再生医学中使用机械和成骨分化培养基的组合来培养 UC-MSCs 具有重要意义。