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大鼠软骨损伤体外模型中波形蛋白细胞骨架对单次冲击负荷的反应变化

Alterations in the vimentin cytoskeleton in response to single impact load in an in vitro model of cartilage damage in the rat.

作者信息

Henson Frances M D, Vincent Thea A

机构信息

Department of Veterinary Medicine, University of Cambridge, Madingley Road, Cambridge, CB3 0ES, UK.

出版信息

BMC Musculoskelet Disord. 2008 Jun 24;9:94. doi: 10.1186/1471-2474-9-94.

Abstract

BACKGROUND

Animal models have provided much information on molecular and cellular changes in joint disease, particularly OA. However there are limitations to in vivo work and single tissue in vitro studies can provide more specific information on individual events. The rat is a commonly used laboratory species but at the current time only in vivo models of rat OA are available to study. The purpose of this study was to investigate the damage that single impact load (SIL) of 0.16J causes in a rat cartilage in vitro model and assess whether this load alters the arrangement of vimentin.

METHODS

Rat cartilage was single impact loaded (200 g from 8 cm) and cultured for up to 48 hours (n = 72 joints). Histological changes were measured using a semi-quantitative modified Mankin score. Immunolocalisation was used to identify changes in vimentin distribution.

RESULTS

SIL caused damage in 32/36 cartilage samples. Damage included surface fibrillation, fissures, fragmentation, changes in cellularity and loss of proteoglycan. SIL caused a statistically significant increase in modified Mankin score and chondrocyte clusters over time. SIL caused vimentin disassembly (as evidenced by collapse of vimentin around the nucleus).

CONCLUSION

This study describes a model of SIL damage to rat cartilage. SIL causes changes in histological/chemical parameters which have been measured using a semi-quantitative modified Mankin score. Single impact load also causes changes in the pattern of vimentin immunoreactivity, indicating vimentin dissassembley. Using a semi-quantitative scoring system the disassembly was shown to be statistically significant in SIL damaged cartilage. The changes described in this paper suggest that this novel single tissue rat model of joint damage is a possible candidate model to replace in vivo models.

摘要

背景

动物模型已提供了许多关于关节疾病,特别是骨关节炎(OA)分子和细胞变化的信息。然而,体内研究存在局限性,单一组织的体外研究可以提供关于个体事件更具体的信息。大鼠是常用的实验动物,但目前仅有大鼠OA的体内模型可供研究。本研究的目的是调查0.16J的单次冲击负荷(SIL)对大鼠软骨体外模型造成的损伤,并评估该负荷是否会改变波形蛋白的排列。

方法

对大鼠软骨施加单次冲击负荷(从8厘米高度落下200克重物),并培养长达48小时(n = 72个关节)。使用半定量改良曼金评分法测量组织学变化。采用免疫定位法确定波形蛋白分布的变化。

结果

SIL导致32/36个软骨样本受损。损伤包括表面原纤维形成、裂隙、碎片形成、细胞数量变化和蛋白聚糖丢失。随着时间的推移,SIL导致改良曼金评分和软骨细胞簇在统计学上显著增加。SIL导致波形蛋白解聚(表现为细胞核周围波形蛋白塌陷)。

结论

本研究描述了一种SIL对大鼠软骨造成损伤的模型。SIL导致组织学/化学参数发生变化,这些变化通过半定量改良曼金评分法进行测量。单次冲击负荷还导致波形蛋白免疫反应模式发生变化,表明波形蛋白解聚。使用半定量评分系统,显示在SIL损伤的软骨中解聚具有统计学意义。本文所述的变化表明,这种新型的单一组织大鼠关节损伤模型可能是替代体内模型的候选模型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f895/2443134/2390d44cc03b/1471-2474-9-94-1.jpg

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