Chondrocyte extracellular matrix synthesis and turnover are influenced by static compression in a new alginate disk culture system
Chondrocyte extracellular matrix synthesis and turnover are influenced by static compression in a new alginate disk culture system
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DOI：
10.1006/abbi.2000.2060
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发表时间：
2000-11-15
影响因子：
3.9
通讯作者：
Sandy, JD
中科院分区：
文献类型：
--
作者：
Ragan, PM;Chin, VI;Sandy, JD
本研究的目的是研究在一种新的藻酸盐培养系统中，机械压缩对软骨细胞生物合成细胞外基质（ECM）成分的影响。具体而言，我们已经研究了软骨细胞的生物合成速率，和存在于细胞相关基质（CM），在进一步去除基质（FRM）和周围的培养基中的聚集蛋白聚糖核心蛋白种类的结构。在这种藻酸盐盘培养系统中，软骨细胞可以经受类似于体内经历的机械变形。我们的研究结果表明，在8周的培养期内，软骨细胞合成功能性ECM，并能对机械力作出反应，类似于保持在天然软骨中的软骨细胞。在藻酸盐盘系统中，静态压缩被证明是减少和动态压缩，以增加牛软骨细胞的聚集蛋白聚糖的合成。蛋白质印迹分析的核心蛋白的聚集蛋白聚糖分子鉴定了许多不同的物种，存在于CM，FRM，和介质中的不同的相对量。在21天的培养中，在ECM中发现的聚集蛋白聚糖的主要形式是全长连接稳定的种类。此外，我们的数据表明，40小时的静态压缩的应用引起的新合成的聚集蛋白聚糖分子释放到介质中的比例增加。然而，这并不伴随着一个显着的变化，聚集蛋白聚糖和聚集蛋白聚糖片段的大小和组成在不同的隔室，这表明机械压缩并没有改变的分解代谢途径。总之，这些数据表明软骨细胞功能在藻酸盐盘培养系统中得以维持，并且该培养系统是检查软骨细胞ECM组装和通常在体内发现的软骨素的某些方面的有用模型，（C）2000 Academic Press。
The goal of this study was to examine the effects of mechanical compression on chondrocyte biosynthesis of extracellular matrix (ECM) components during culture in a new alginate disk culture system. Specifically, we have examined chondrocyte biosynthesis rates, and the structure of aggrecan core protein species present in the cell-associated matrix (CM), in the further removed matrix (FRM) and in the surrounding culture medium. In this alginate disk culture system, chondrocytes can be subjected to mechanical deformations similar to those experienced in vivo. Our results show that over an 8-week culture period, chondrocytes synthesize a functional ECM and can respond to mechanical forces similarly to chondrocytes maintained in native cartilage. In the alginate disk system, static compression was shown to decrease and dynamic compression to increase synthesis of aggrecan of bovine chondrocytes. Western blot analysis of the core proteins of aggrecan molecules identified a number of different species that were present in different relative amounts in the CM, FRM, and medium. Over 21 days of culture, the predominant form of aggrecan found in the ECM was a full-length link-stabilized species. In addition, our data show that the application of 40 h of static compression caused an increase in the proportion of newly synthesized aggrecan molecules released into the medium. However, this was not accompanied by a significant change in the size and composition of aggrecan and aggrecan fragments in the different compartments, suggesting that mechanical compression did not alter the catabolic pathways. Together, these data show that chondrocyte function is maintained in an alginate disk culture system and that this culture system is a useful model to examine chondrocyte ECM assembly and some aspects of catabolism normally found in vivo, (C) 2000 Academic Press.