Sharma G, Saxena R K, Mishra P
Centre for Biomedical Engineering, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India.
Osteoarthritis Cartilage. 2008 Nov;16(11):1387-94. doi: 10.1016/j.joca.2008.03.026. Epub 2008 May 16.
To investigate the synergistic effect of chondroitin sulfate (CS) and cyclic pressure on the biochemical and morphological properties of chondrocytes isolated from articular cartilage and cultured in alginate matrix.
The chondrocytes obtained from articular cartilage of goat femoropatellar joint were isolated and cultured in alginate matrix. The cells were exposed to CS (100 microg/ml) along with cyclic pressure of 1.2 MPa and 2.4 MPa and biochemical analysis of DNA, proteoglycan, collagen and protease activity was carried out in different matrix fractions, i.e., cellular matrix (CM) and further removed matrix (FRM) and in culture medium. The morphological studies of chondrocytes were carried out using transmission electron microscopy (TEM).
The treatment of chondrocytes with CS along with cyclic pressure increased the rate of cell proliferation relative to control (without load and in the absence of CS) and CS alone (P<0.001). The proteoglycan content in CM increased in the presence of CS alone (P<0.05) as well as CS with cyclic pressure (P<0.001). The specific activity of protease in CM and FRM decreased in the presence of CS with cyclic pressure relative to control (P<0.001). The TEM images showed abundant CM, improved cell morphology and matrix organization in the presence of CS and cyclic load treatment.
The results of this study suggested that in the presence of CS along with cyclic loading, the cellular ability to utilize and incorporate exogenous CS as extracellular matrix improved, as compared to CS alone.
研究硫酸软骨素(CS)与循环压力对从关节软骨分离并在藻酸盐基质中培养的软骨细胞的生化和形态学特性的协同作用。
从山羊股骨髌骨关节的关节软骨中获取软骨细胞,在藻酸盐基质中分离并培养。将细胞暴露于CS(100微克/毫升)以及1.2兆帕和2.4兆帕的循环压力下,对不同基质部分,即细胞基质(CM)和进一步去除的基质(FRM)以及培养基中的DNA、蛋白聚糖、胶原蛋白和蛋白酶活性进行生化分析。使用透射电子显微镜(TEM)对软骨细胞进行形态学研究。
与对照组(无负荷且无CS)和单独使用CS相比,CS联合循环压力处理软骨细胞可提高细胞增殖率(P<0.001)。单独使用CS时(P<0.05)以及CS联合循环压力时(P<0.001),CM中的蛋白聚糖含量增加。与对照组相比,CS联合循环压力时,CM和FRM中蛋白酶的比活性降低(P<0.001)。TEM图像显示,在CS和循环负荷处理下,CM丰富,细胞形态和基质组织得到改善。
本研究结果表明,与单独使用CS相比,在CS联合循环加载的情况下,细胞利用和整合外源性CS作为细胞外基质的能力得到了提高。