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在体和体外对小鼠跟腱和腱细胞的机械加载:一项初步研究。

In Vivo and In Vitro Mechanical Loading of Mouse Achilles Tendons and Tenocytes-A Pilot Study.

机构信息

Department of Trauma-, Hand- and Reconstructive Surgery, Experimental Trauma Surgery, University Hospital Jena, 07743 Jena, Germany.

Julius Wolff Institute, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 13353 Berlin, Germany.

出版信息

Int J Mol Sci. 2020 Feb 15;21(4):1313. doi: 10.3390/ijms21041313.

Abstract

Mechanical force is a key factor for the maintenance, adaptation, and function of tendons. Investigating the impact of mechanical loading in tenocytes and tendons might provide important information on in vivo tendon mechanobiology. Therefore, the study aimed at understanding if an in vitro loading set up of tenocytes leads to similar regulations of cell shape and gene expression, as loading of the Achilles tendon in an in vivo mouse model. In vivo: The left tibiae of mice ( = 12) were subject to axial cyclic compressive loading for 3 weeks, and the Achilles tendons were harvested. The right tibiae served as the internal non-loaded control. In vitro: tenocytes were isolated from mice Achilles tendons and were loaded for 4 h or 5 days ( = 6 per group) based on the in vivo protocol. Histology showed significant differences in the cell shape between in vivo and in vitro loading. On the molecular level, quantitative real-time PCR revealed significant differences in the gene expression of collagen type I and III and of the matrix metalloproteinases (MMP). Tendon-associated markers showed a similar expression profile. This study showed that the gene expression of tendon markers was similar, whereas significant changes in the expression of extracellular matrix (ECM) related genes were detected between in vivo and in vitro loading. This first pilot study is important for understanding to which extent in vitro stimulation set-ups of tenocytes can mimic in vivo characteristics.

摘要

力学刺激是维持、适应和发挥肌腱功能的关键因素。研究力学加载对肌腱细胞和肌腱的影响可能为体内肌腱生物力学提供重要信息。因此,本研究旨在探讨体外肌腱细胞加载是否会导致类似的细胞形态和基因表达调节,如同在体内小鼠模型中对跟腱进行加载一样。

  • 体内实验:12 只小鼠的左侧胫骨接受轴向循环压缩加载 3 周,并采集跟腱。右侧胫骨作为内部非加载对照。

  • 体外实验:从小鼠跟腱中分离肌腱细胞,并根据体内方案进行 4 小时或 5 天的加载(每组 6 只)。组织学显示体内和体外加载的细胞形态存在显著差异。在分子水平上,实时定量 PCR 显示Ⅰ型和Ⅲ型胶原以及基质金属蛋白酶 (MMP) 的基因表达存在显著差异。与肌腱相关的标志物表现出相似的表达模式。

本研究表明,肌腱标志物的基因表达相似,但在体内和体外加载之间检测到细胞外基质 (ECM) 相关基因的表达有显著变化。这项初步的试点研究对于了解体外肌腱细胞刺激装置在多大程度上可以模拟体内特征非常重要。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff5a/7072865/9fc1f8b8a913/ijms-21-01313-g001.jpg

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