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单细胞对机械力反应的分子谱分析:软骨细胞、软骨小体和包封软骨细胞的比较。

Molecular profiling of single cells in response to mechanical force: comparison of chondrocytes, chondrons and encapsulated chondrocytes.

机构信息

Institute for Science & Technology in Medicine, University of Keele, Staffordshire ST5 5BG, UK.

出版信息

Biomaterials. 2010 Mar;31(7):1619-25. doi: 10.1016/j.biomaterials.2009.11.021. Epub 2009 Dec 1.

DOI:10.1016/j.biomaterials.2009.11.021
PMID:19954841
Abstract

A chondrocyte and its surrounding pericellular matrix (PCM) are defined as a chondron. The PCM plays a critical role in enhancing matrix production, protecting the chondrocyte during loading and transducing mechanical signals. Tissue engineering involves the design of artificial matrices which aim to mimic PCM function for mechanical strength and signalling motifs. We compare the mechanical performance and mechanoresponsiveness of chondrocytes with and without PCM, and encapsulated by alternate adsorption of two oppositely charged polyelectrolytes; chitosan and hyaluronan. Zeta potential measurements confirmed the success of the encapsulation. Encapsulation did not influence chondrocyte viability or metabolic activity. Cells were compressed by micromanipulation with final deformations to 30%, 50% and 70%. Force-displacement data showed that the larger the deformation at the end of compression, the greater the force on the cell. Mechanoresponsiveness of cells was studied by combining single cell PCR with dynamic compression at 20% and 40%. Aggrecan and Type II collagen gene expression were significantly increased in encapsulated chondrocytes and chondrons compared to chondrocytes whereas dynamic compression had no effect on SOX9 or lubricin gene expression. Our results demonstrate that although encapsulation can mimic responses of chondrocytes to biomechanical compression the molecular profile did not reach the enhanced levels observed with chondrons.

摘要

软骨细胞及其周围的细胞外基质(PCM)被定义为软骨小体。PCM 在增强基质生成、在加载过程中保护软骨细胞以及传递机械信号方面起着关键作用。组织工程涉及到人工基质的设计,旨在模仿 PCM 的功能,以获得机械强度和信号基序。我们比较了有和没有 PCM 的软骨细胞以及通过两种带相反电荷的聚电解质交替吸附包封的软骨细胞的机械性能和机械响应性;壳聚糖和透明质酸。Zeta 电位测量证实了封装的成功。封装不影响软骨细胞的活力或代谢活性。通过微操作对细胞进行压缩,最终压缩变形为 30%、50%和 70%。力-位移数据表明,压缩结束时的变形越大,细胞上的力就越大。通过在 20%和 40%的动态压缩下结合单细胞 PCR 研究细胞的机械响应性。与软骨细胞相比,包封的软骨细胞和软骨小体中的聚集蛋白聚糖和 II 型胶原基因表达显著增加,而动态压缩对 SOX9 或润滑素基因表达没有影响。我们的结果表明,尽管封装可以模拟软骨细胞对生物力学压缩的反应,但分子谱没有达到与软骨小体观察到的增强水平。

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