Response of Articular Cartilage to Hyperosmolar Stress: Report of an Ex Vivo Injury Model

Clemens Unterguggenberger, Shahrbanoo Jahangir, Gian M. Salzmann, Martin J. Stoddart, Sibylle Grad, Hagen Schmal, Eva Johanna Kubosch
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Abstract

Background:Physiological 0.9% saline is commonly used as an irrigation fluid in modern arthroscopy. There is a growing body of evidence that a hyperosmolar saline solution has chondroprotective effects, especially if iatrogenic injury occurs.Purpose:To (1) corroborate the superiority of a hyperosmolar saline solution regarding chondrocyte survival after mechanical injury and (2) observe the modulatory response of articular cartilage to osmotic stress and injury.Study Design:Controlled laboratory study.Methods:Osteochondral explants were isolated from bovine stifle joints and exposed to either 0.9% saline (308 mOsm) or hyperosmolar saline (600 mOsm) and then damaged with a sharp dermatome blade to attain a confined full-thickness cartilage injury site, incubated in the same fluids for another 3 hours, and transferred to chondropermissive medium for further culture for 1 week. Chondrocyte survival was assessed by confocal imaging, while the cellular response was evaluated over 1 week by relative gene expression for apoptotic and inflammatory markers and mediator release into the medium.Results:The full-thickness cartilage cut resulted in a confined zone of cell death that mainly affected superficial zone chondrocytes. Injured samples that were exposed to hyperosmolar saline showed less expansion of cell death in both the axial ( P < .007) and the coronal ( P < .004) plane. There was no progression of cell death during the following week of culture. Histological assessment revealed an intact cartilage matrix and normal chondrocyte morphology. Inflammatory and proapoptotic genes were upregulated on the first days postexposure with a notable downregulation toward day 7. Mediator release into the medium was concentrated on day 3.Conclusion:This in vitro cartilage injury model provides further evidence for the chondroprotective effect of a hyperosmolar saline irrigation fluid, as well as novel data on the capability of articular cartilage to quickly regain joint homeostasis after osmotic stress and injury.Clinical Relevance:Raising the osmolarity of an irrigating solution may be a simple and safe strategy to protect articular cartilage during arthroscopic surgery.
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关节软骨对高渗压力的反应:体内外损伤模型报告
背景:生理盐水 0.9% 通常用作现代关节镜手术的冲洗液。目的:(1) 证实高渗盐水在机械损伤后软骨细胞存活方面的优越性;(2) 观察关节软骨对渗透压和损伤的调节反应。研究方法:从牛跗关节中分离出骨软骨外植体,将其暴露于 0.9% 生理盐水(308 mOsm)或高渗透压生理盐水(600 mOsm)中,然后用锋利的皮膜刀片对其进行损伤,以获得封闭的全厚软骨损伤部位,在相同的液体中再培养 3 小时,然后转移到软骨渗透培养基中继续培养 1 周。通过共焦成像评估软骨细胞的存活率,同时通过细胞凋亡和炎症标志物的相对基因表达以及介质在培养基中的释放来评估细胞在一周内的反应。暴露于高渗盐水的损伤样本在轴向(P < .007)和冠状面(P < .004)上的细胞死亡扩展较少。在随后一周的培养过程中,细胞死亡没有继续发展。组织学评估显示软骨基质完好无损,软骨细胞形态正常。炎症基因和促凋亡基因在暴露后的头几天上调,在第 7 天明显下调。结论:这一体外软骨损伤模型进一步证明了高渗透压生理盐水冲洗液的软骨保护作用,并提供了关节软骨在受到渗透压和损伤后迅速恢复关节平衡能力的新数据。
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