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Biophys J, March 2000, p. 1578-1588, Vol. 78, No. 3

Scanning Electrochemical Microscopy as a Local Probe of Oxygen Permeability in Cartilage

Marylou Gonsalves,* Anna L. Barker,* Julie V. Macpherson,* Patrick R. Unwin,* Danny O'Hare,dagger and C. Peter WinloveDagger

 *Department of Chemistry, University of Warwick, Coventry CV4 7AL;  dagger Department of Pharmacy, University of Brighton, Brighton BN2 4GJ; and  Dagger Physiological Flow Studies Group, Department of Biological and Medical Systems, Imperial College of Science Technology and Medicine, London SW7 2BY, United Kingdom

The use of scanning electrochemical microscopy, a high-resolution chemical imaging technique, to probe the distribution and mobility of solutes in articular cartilage is described. In this application, a mobile ultramicroelectrode is positioned close (~1 µm) to the cartilage sample surface, which has been equilibrated in a bathing solution containing the solute of interest. The solute is electrolyzed at a diffusion-limited rate, and the current response measured as the ultramicroelectrode is scanned across the sample surface. The topography of the samples was determined using Ru(CN)64-, a solute to which the cartilage matrix was impermeable. This revealed a number of pit-like depressions corresponding to the distribution of chondrocytes, which were also observed by atomic force and light microscopy. Subsequent imaging of the same area of the cartilage sample for the diffusion-limited reduction of oxygen indicated enhanced, but heterogeneous, permeability of oxygen across the cartilage surface. In particular, areas of high permeability were observed in the cellular and pericellular regions. This is the first time that inhomogeneities in the permeability of cartilage toward simple solutes, such as oxygen, have been observed on a micrometer scale.

Biophys J, March 2000, p. 1578-1588, Vol. 78, No. 3
© 2000 by the Biophysical Society   0006-3495/00/03/1578/11  $2.00


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