Kornicka Katarzyna, Al Naem Mohamad, Röcken Michael, Zmiertka Marta, Marycz Krzysztof
International Institute of Translational Medicine, Jesionowa, 11, Malin, 55-114 Wisznia Mała, Poland.
Department of Experimental Biology, Wroclaw University of Environmental and Life Sciences, 50-375 Wroclaw, Poland.
J Clin Med. 2019 Mar 8;8(3):328. doi: 10.3390/jcm8030328.
Osteochondritis dissecans (OCD) in equids, especially in sport horses, has become a growing issue as it contributes to the occurrence of lameness. Thus the aim of this study was to investigate the cytophysiological properties of OCD chondrocytes including expression of chondrogenic genes, apoptosis, mitochondria dynamics and autophagy. Horse chondrocytes were isolated from healthy (HE) and OCD cartilages. Properties of cells were evaluated using multiple assays e.g., polymerase chain reaction (PCR), immunofluorescence, Western blot. OCD chondrocytes were characterized by increased apoptosis and senescence. Expression of chondrogenic genes (vimentin, aggrecan) was decreased while mRNA levels of matrix metalloproteinase 13 significantly upregulated in comparison to HE cells. Moreover, OCD cells displayed increased mitochondrial fusion while fission events were diminished. Interestingly, chaperone mediated autophagy was triggered in those cells and it predominated over macroautophagy. Furthermore, co-culture of LPS-treated chondrocytes with adipose-derived stem cells (ASC) decreased p62/sequestosome 1 (SQSTM) and increases MMP-13 expression in OCD cells. Our results suggest that OCD affected horse chondrocytes are characterized by senescent phenotype due to endoplasmic reticulum stress and mitochondria dynamics deterioration. Expression of chondrogenic markers is decreased in those cells while expression of chaperone mediated autophagy (CMA)-related genes increased. Increased malfunctioning of cells leads to loss of their functionality and capacity to maintain tissue homeostasis.
马的剥脱性骨软骨炎（OCD），尤其是在运动马中，已成为一个日益严重的问题，因为它会导致跛行的发生。因此，本研究的目的是研究OCD软骨细胞的细胞生理特性，包括软骨生成基因的表达、凋亡、线粒体动力学和自噬。从健康（HE）和OCD软骨中分离出马软骨细胞。使用多种检测方法评估细胞特性，例如聚合酶链反应（PCR）、免疫荧光、蛋白质免疫印迹法。OCD软骨细胞的特征是凋亡和衰老增加。与HE细胞相比，软骨生成基因（波形蛋白、聚集蛋白聚糖）的表达降低，而基质金属蛋白酶13的mRNA水平显著上调。此外，OCD细胞的线粒体融合增加，而裂变事件减少。有趣的是，伴侣介导的自噬在这些细胞中被触发，并且它比巨自噬占主导地位。此外，用脂多糖处理的软骨细胞与脂肪来源的干细胞（ASC）共培养可降低OCD细胞中p62/聚集体蛋白1（SQSTM）的水平并增加MMP-13的表达。我们的结果表明，受OCD影响的马软骨细胞具有衰老表型，这是由于内质网应激和线粒体动力学恶化所致。这些细胞中软骨生成标志物的表达降低，而伴侣介导的自噬（CMA）相关基因的表达增加。细胞功能障碍的增加导致其功能丧失以及维持组织稳态的能力丧失。
