Key Laboratory for Space Biosciences and Biotechnology, School of Life Sciences, Northwestern Polytechnical University, Xi'an, 710072, Shaanxi, China.
Department of Biochemistry, University of Texas Health Science Center at San Antonio, San Antonio, TX, 78229, USA.
J Bone Miner Metab. 2018 Sep;36(5):529-536. doi: 10.1007/s00774-017-0870-3. Epub 2017 Oct 12.
This study constructed an in situ cell culture, real-time observation system based originally on a microfluidic channel, and reported the morphological changes of late osteoblast-like IDG-SW3 cells in response to flow shear stress (FSS). The effects of high (1.2 Pa) and low (0.3 Pa) magnitudes of unidirectional FSS and three concentrations of extracellular Type I collagen (0.1, 0.5, and 1 mg/mL) coating on cell morphology were investigated. IDG-SW3 cells were cultured in polydimethylsiloxane microfluidic channels. Cell images were recorded real-time under microscope at intervals of 1 min. Cell morphology was characterized by five parameters: cellular area, cell elongation index, cellular alignment, cellular process length, and number of cellular process per cell. Immunofluorescence assay was used to detect stress fiber distribution and vinculin expression. The results showed that 1.2 Pa, but not 0.3 Pa of FSS induced a significant morphological change in late osteoblast-like IDG-SW3 cells, which may be caused by the alteration of cellular adhesion with matrix in response to FSS. Moreover, the amount of collagen matrix, alignment of fiber stress and expression of vinculin were closely correlated with the morphological changes of IDG-SW3 cells. This study suggests that osteoblasts are very responsive to the magnitudes of FSS, and extracellular collagen matrix and focal adhesion are directly involved in the morphological changes adaptive to FSS.
本研究构建了一种基于微流控通道的原位细胞培养实时观察系统,并报道了成骨样 IDG-SW3 细胞对流动切应力(FSS)的形态变化。研究了高(1.2 Pa)、低(0.3 Pa)单向 FSS 幅度和三种浓度细胞外 I 型胶原(0.1、0.5 和 1 mg/mL)涂层对细胞形态的影响。IDG-SW3 细胞在聚二甲基硅氧烷微流控通道中培养。每隔 1 分钟在显微镜下实时记录细胞图像。通过五个参数描述细胞形态:细胞面积、细胞伸长指数、细胞取向、细胞突起长度和每个细胞的细胞突起数量。免疫荧光染色检测应力纤维分布和粘着斑蛋白表达。结果表明,1.2 Pa 的 FSS 而非 0.3 Pa 的 FSS 诱导晚期成骨样 IDG-SW3 细胞发生显著的形态变化,这可能是由于细胞与基质的黏附在 FSS 作用下发生改变所致。此外,胶原基质的量、纤维应力的取向和粘着斑蛋白的表达与 IDG-SW3 细胞的形态变化密切相关。本研究表明,成骨细胞对 FSS 的幅度非常敏感,细胞外胶原基质和黏着斑直接参与了对 FSS 的形态适应性变化。