School of Engineering and Materials Science, Queen Mary University of London, London, UK.
J Cell Physiol. 2010 Jun;223(3):630-9. doi: 10.1002/jcp.22061.
In the absence of in vivo measurements, the oxygen concentration within articular cartilage is calculated from the balance between cellular oxygen consumption and mass transfer. Current estimates of the oxygen tension within articular cartilage are based on oxygen consumption data from full-depth tissue samples. However, superficial and deep cell subpopulations of articular cartilage express intrinsic metabolic differences. We test the hypothesis that the subpopulations differ with respect to their intrinsic oxygen consumption rate. Chondrocytes from the full cartilage thickness demonstrate enhanced oxygen consumption when deprived of glucose, consistent with the Crabtree phenomena. Chondrocyte subpopulations differ in the prevailing availability of oxygen and glucose, which decrease with distance from the cartilage-synovial fluid interface. Thus, we tested the hypothesis that the oxygen consumption of each subpopulation is modulated by nutrient availability, by examining the expression of the Crabtree effect. The deep cells had a greater oxygen consumption than the superficial cells (V(max) of 6.6 compared to 3.2 fmol/cell/h), consistent with our observations of mitochondrial volume (mean values 52.0 vs. 36.4 microm(3)/cell). Both populations expressed the Crabtree phenomena, with oxygen consumption increasing approximately 2.5-fold in response to glycolytic inhibition by glucose deprivation or 2-deoxyglucose. Over 90% of this increase was oligomycin-sensitive and thus accounted for by oxidative phosphorylation. The data contributes towards our understanding of chondrocyte energy metabolism and provides information valuable for the accurate calculation of the oxygen concentration that the cells experience in vivo. The work has further application to the optimisation of bioreactor design and engineered tissues.
在缺乏体内测量的情况下,关节软骨内的氧浓度是通过细胞耗氧量和质量转移之间的平衡来计算的。目前对关节软骨内氧张力的估计是基于来自全层组织样本的耗氧数据。然而,关节软骨的浅层和深层细胞亚群表现出内在代谢的差异。我们检验了这样一个假设,即亚群在其内在耗氧率方面存在差异。从整个软骨厚度获得的软骨细胞在剥夺葡萄糖时表现出增强的耗氧量,这与 Crabtree 现象一致。软骨细胞亚群在氧气和葡萄糖的可利用性方面存在差异,而这些物质随着与软骨-滑液界面距离的增加而减少。因此,我们通过检查 Crabtree 效应的表达,测试了每个亚群的耗氧量是否受到营养物质可用性的调节这一假设。深层细胞的耗氧量大于浅层细胞(V(max)分别为 6.6 和 3.2 fmol/细胞/h),与我们观察到的线粒体体积(平均值分别为 52.0 和 36.4 µm³/细胞)一致。两个群体都表达了 Crabtree 现象,在葡萄糖剥夺或 2-脱氧葡萄糖抑制糖酵解时,耗氧量增加约 2.5 倍。超过 90%的增加是寡霉素敏感的,因此归因于氧化磷酸化。该数据有助于我们理解软骨细胞的能量代谢,并提供了有价值的信息,有助于准确计算细胞在体内经历的氧浓度。这项工作进一步应用于生物反应器设计和工程组织的优化。