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TECHNICAL PAPERS: Cell

Nutrient Utilization by Bovine Articular Chondrocytes: A Combined Experimental and Theoretical Approach

[+] Author and Article Information
Bram G. Sengers, Cees W. Oomens

 Eindhoven University of Technology, Department of Biomedical Engineering, P.O. Box 513, 5600 MB Eindhoven, The Netherlands

Hannah K. Heywood, David A. Lee

 Queen Mary University of London, Medical Engineering and IRC Biomedical Materials, Department of Engineering, Mile End Road, London E1 4NS, United Kingdom

Dan L. Bader

 Eindhoven University of Technology, Department of Biomedical Engineering, P.O. Box 513, 5600 MB Eindhoven, The Netherlands and  Queen Mary University of London, Medical Engineering and IRC Biomedical Materials, Department of Engineering, Mile End Road, London E1 4NS, United Kingdom

J Biomech Eng 127(5), 758-766 (May 26, 2005) (9 pages) doi:10.1115/1.1993664 History: Received December 22, 2004; Revised May 26, 2005

A combined experimental-numerical approach was adopted to characterize glucose and oxygen uptake and lactate production by bovine articular chondrocytes in a model system. For a wide range of cell concentrations, cells in agarose were supplemented with either low or high glucose medium. During an initial culture phase of 48h, oxygen was monitored noninvasively using a biosensor system. Glucose and lactate were determined by medium sampling. In order to quantify glucose and oxygen uptake, a finite element approach was adopted to describe diffusion and uptake in the experimental model. Numerical predictions of lactate, based on simple relations for cell metabolism, were found to agree well for low glucose, but not for high glucose medium. Oxygen did not play a role in either case. Given the close association between chondrocyte energy metabolism and matrix synthesis, a quantifiable prediction of utilization can present a valuable contribution in the optimization of tissue engineering conditions.

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Copyright © 2005 by American Society of Mechanical Engineers
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Figures

Grahic Jump Location
Figure 1

Geometry of an individual well of the 96-well plate setup. An oxygen sensitive fluorphore is immobilized in a silicon matrix at the well base. 65μL of cells embedded in 4% agarose was added and supplemented with 200μL culture medium, giving a total diffusion distance of 8.55mm from the free surface to the oxygen sensor.

Grahic Jump Location
Figure 2

Low glucose. (a) Glucose measurements and the computed mean medium glucose concentration, as a function of time, for 1.25–40×106cellcm−3. (b) Computed spatial glucose distribution in the well for 40×106cellcm−3, at 1h intervals. The free surface is located to the left and the well base to the right. The medium zone extents from 0to6.45mm and the agarose zone from 6.45to8.55mm.

Grahic Jump Location
Figure 3

High glucose. (a) Glucose measurements and the computed mean medium glucose concentration, as a function of time, for 1.25–40×106cellcm−3. (b) Computed spatial glucose distribution in the well for 40×106cellcm−3, at 1h intervals. The free surface is located to the left and the well base to the right. The medium zone extents from 0to6.45mm and the agarose zone from 6.45to8.55mm.

Grahic Jump Location
Figure 4

Low glucose. (a) Lactate measurements and the predicted mean medium lactate concentration based on glucose alone Eq. 3, 1.25–40×106cellcm−3. (b) Computed spatial lactate distribution in the well for 40×106cellcm−3, at 1h intervals. The free surface is located to the left and the well base to the right.

Grahic Jump Location
Figure 5

High glucose. (a) Lactate measurements and the predicted mean medium lactate concentration based on glucose alone, Eq. 3, 1.25–40×106cellcm−3. (b) Computed spatial lactate distribution in the well for 40×106cellcm−3, at 1h intervals. The free surface is located to the left and the well base to the right.

Grahic Jump Location
Figure 6

Low glucose. (a) Measured and computed base well oxygen concentration, 1.25–40×106cellcm−3. (b) Computed spatial oxygen distribution in the well for 40×106cellcm−3, at 1h intervals. The free surface is located to the left and the well base to the right.

Grahic Jump Location
Figure 7

High glucose. (a) Measured and computed base well oxygen concentration, 1.25–40×106cellcm−3. (b) Computed spatial oxygen distribution in the well for 40×106cellcm−3, at 1h intervals. The free surface is located to the left and the well base to the right.

Grahic Jump Location
Figure 8

Lactate measurements and the predicted mean medium lactate concentration based on glucose and oxygen, Eq. 4, 1.25–40×106cellcm−3. (a) Low glucose. (b) High glucose.

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