Regional cell density distribution and oxygen consumption rates in porcine TMJ discs: an explant study J. Kuo, C. Shi, S. Cisewski, L. Zhang, M.J. Kern,

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Regional cell density distribution and oxygen consumption rates in porcine TMJ discs: an explant study J. Kuo, C. Shi, S. Cisewski, L. Zhang, M.J. Kern, H. Yao Osteoarthritis and Cartilage Volume 19, Issue 7, Pages 911-918 (July 2011) DOI: 10.1016/j.joca.2011.03.002 Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 (A) Representation of the regions investigated for determining cell density and oxygen consumption rate. The TMJ disc was divided into five regions: anterior, intermediate, posterior, lateral and medial. From each region, sections (100μm) were further taken from three layers for cell density measurement. (B) Schematic of 3D cell counting using a confocal microscopy technique. Osteoarthritis and Cartilage 2011 19, 911-918DOI: (10.1016/j.joca.2011.03.002) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 (A) Schematic of the experimental setup for oxygen consumption rate measurements. TMJ disc explants were cultured in a sealed water-jacketed metabolism chamber. (B) Typical record of dissolved oxygen concentration in the chamber over time. The experimental data were curve fit to Equation (3). (C) The oxygen consumption rate was plotted based on the Michaelis–Menten equation with determined parameters Vmax and Km. Osteoarthritis and Cartilage 2011 19, 911-918DOI: (10.1016/j.joca.2011.03.002) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Comparison of cell density between the confocal microscopy technique (n=5) and DNA assay (n=5). The error bar is a 95% CI. No significant differences were detected in each disc region. Osteoarthritis and Cartilage 2011 19, 911-918DOI: (10.1016/j.joca.2011.03.002) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 (A) Relationship between the oxygen consumption rate and the oxygen tension based on the Michaelis–Menten equation with averaged cell based Vmax and Km in five disc regions. (B) Predicted relationship between the relative oxygen consumption rate and the oxygen tension. The averaged Km across the TMJ disc was 19.1μmol/L. The oxygen consumption rate was normalized by the rate at 21% oxygen. Osteoarthritis and Cartilage 2011 19, 911-918DOI: (10.1016/j.joca.2011.03.002) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions