Effect of age and cytoskeletal elements on the indentation-dependent mechanical properties of chondrocytes.
Effect of age and cytoskeletal elements on the indentation-dependent mechanical properties of chondrocytes.
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DOI：
10.1371/journal.pone.0061651
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Loots GG
中科院分区：
文献类型：
--
作者：
Chahine NO;Blanchette C;Thomas CB;Lu J;Haudenschild D;Loots GG
关节软骨软骨细胞负责细胞外基质的合成、维持和周转，代谢过程有助于这些细胞的机械特性。在这里，我们系统地评估了年龄和细胞骨架破坏物对软骨细胞变形的力学性能的影响。我们量化的压痕依赖的机械性能的软骨细胞分离新生儿（1天），成人（5年）和老年（12年）牛膝关节使用原子力显微镜（AFM）。我们还测量了肌动蛋白和中间丝的软骨细胞的压痕依赖的机械性能的贡献。通过将AFM与共聚焦荧光显微镜相结合，我们监测了转基因细胞（GFP-波形蛋白和mCherry-肌动蛋白）在压缩下的细胞骨架和生物力学变形。我们发现，在所有年龄组的软骨细胞的弹性模量随着压痕（15-2000 nm）的增加而降低。在大于500 nm的压痕处，成人软骨细胞的弹性模量显著大于新生细胞。粘弹性模量（瞬时和平衡）在所有年龄组的检查是可比的，但是，固有粘度低于老年软骨细胞比新生儿。破坏肌动蛋白或中间丝结构改变了软骨细胞的机械性能，通过降低弹性模量和粘弹性，导致在一个显着的损失与治疗的压痕依赖性的反应。肌动蛋白和波形蛋白的细胞骨架结构进行了监测，使用共聚焦荧光显微镜在转基因细胞处理的干扰物，这两种治疗方法有一个深刻的破坏性影响的肌动蛋白丝。在这里，我们表明，破坏中间丝的结构间接改变了肌动蛋白细胞骨架的配置。这些发现强调了软骨细胞的整体机械响应中的细胞骨架元素的重要性，表明中间丝的完整性是软骨细胞的非线性弹性特性的关键。本研究在单细胞水平上提高了我们对关节软骨力学特性的理解。
Articular cartilage chondrocytes are responsible for the synthesis, maintenance, and turnover of the extracellular matrix, metabolic processes that contribute to the mechanical properties of these cells. Here, we systematically evaluated the effect of age and cytoskeletal disruptors on the mechanical properties of chondrocytes as a function of deformation. We quantified the indentation-dependent mechanical properties of chondrocytes isolated from neonatal (1-day), adult (5-year) and geriatric (12-year) bovine knees using atomic force microscopy (AFM). We also measured the contribution of the actin and intermediate filaments to the indentation-dependent mechanical properties of chondrocytes. By integrating AFM with confocal fluorescent microscopy, we monitored cytoskeletal and biomechanical deformation in transgenic cells (GFP-vimentin and mCherry-actin) under compression. We found that the elastic modulus of chondrocytes in all age groups decreased with increased indentation (15–2000 nm). The elastic modulus of adult chondrocytes was significantly greater than neonatal cells at indentations greater than 500 nm. Viscoelastic moduli (instantaneous and equilibrium) were comparable in all age groups examined; however, the intrinsic viscosity was lower in geriatric chondrocytes than neonatal. Disrupting the actin or the intermediate filament structures altered the mechanical properties of chondrocytes by decreasing the elastic modulus and viscoelastic properties, resulting in a dramatic loss of indentation-dependent response with treatment. Actin and vimentin cytoskeletal structures were monitored using confocal fluorescent microscopy in transgenic cells treated with disruptors, and both treatments had a profound disruptive effect on the actin filaments. Here we show that disrupting the structure of intermediate filaments indirectly altered the configuration of the actin cytoskeleton. These findings underscore the importance of the cytoskeletal elements in the overall mechanical response of chondrocytes, indicating that intermediate filament integrity is key to the non-linear elastic properties of chondrocytes. This study improves our understanding of the mechanical properties of articular cartilage at the single cell level.
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影响因子：
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