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使用一种新型动态力学测试方法对整个小鼠椎间盘的粘弹性行为进行高通量表型分析。

The high-throughput phenotyping of the viscoelastic behavior of whole mouse intervertebral discs using a novel method of dynamic mechanical testing.

作者信息

Liu Jennifer W, Abraham Adam C, Tang Simon Y

机构信息

Department of Biomedical Engineering, Washington University in St. Louis, St. Louis, MO, United States.

Department of Orthopaedic Surgery, Washington University in St. Louis, 660 South Euclid Avenue, Box 8233, St. Louis, MO 63110, United States.

出版信息

J Biomech. 2015 Jul 16;48(10):2189-94. doi: 10.1016/j.jbiomech.2015.04.040. Epub 2015 May 6.

Abstract

Intervertebral disc (IVD) degeneration is highly correlated with lower back pain, and thus understanding the mechanisms of IVD degeneration is critical for the treatment of this disease. Utilizing mouse models to probe the mechanisms of degeneration is especially attractive due to the ease of manipulating mouse models and the availability of transgenics. Yet characterizing the mechanical behavior of mice IVDs remain challenging due to their minute size (approximately 540 μm in height and 1080 μm(2) in cross sectional area). We have thus developed a simple method to dynamically characterize the mechanical properties of intact mouse IVDs. The IVDs were dissected with the endplates intact, and dynamically compressed in the axial direction at 1% and 5% peak strains at 1 Hz. Utilizing this novel approach, we examined the effects of in vitro ribosylation and trypsin digestion for 24 or 72 h on the viscoelastic behavior of the whole murine IVD. Trypsin treatment resulted in a decrease of proteoglycans and loss of disc height, while ribosylation had no effect on structure or proteoglycan composition. The 72 h ribosylation group exhibited a stiffening of the disc, and both treatments significantly reduced viscous behavior of the IVDs, with the effects being more pronounced at 5% strain. Here we demonstrate a novel high-throughput method to mechanically characterize murine IVDs and detect strain-dependent differences in the elastic and the viscous behavior of the treated IVDs due to ribose and trypsin treatments.

摘要

椎间盘(IVD)退变与下腰痛高度相关,因此了解IVD退变机制对于该疾病的治疗至关重要。利用小鼠模型探究退变机制特别具有吸引力,因为小鼠模型易于操作且有转基因可用。然而,由于小鼠IVD尺寸微小(高度约540μm,横截面积1080μm²),表征其力学行为仍然具有挑战性。因此,我们开发了一种简单方法来动态表征完整小鼠IVD的力学性能。完整保留终板对IVD进行解剖,并在轴向以1Hz的频率在1%和5%峰值应变下进行动态压缩。利用这种新方法,我们研究了体外核糖基化和胰蛋白酶消化24或72小时对整个小鼠IVD粘弹性行为的影响。胰蛋白酶处理导致蛋白聚糖减少和椎间盘高度降低,而核糖基化对结构或蛋白聚糖组成没有影响。72小时核糖基化组的椎间盘出现硬化,两种处理均显著降低了IVD的粘性行为,在5%应变时效果更明显。在这里,我们展示了一种新颖的高通量方法,用于机械表征小鼠IVD,并检测由于核糖和胰蛋白酶处理导致的经处理IVD弹性和粘性行为的应变依赖性差异。

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