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J Biol Chem, Vol. 275, Issue 9, 6321-6327, March 3, 2000

The Organization of Aggrecan in Human Articular Cartilage
EVIDENCE FOR AGE-RELATED CHANGES IN THE RATE OF AGGREGATION OF NEWLY SYNTHESIZED MOLECULES^*

Michael T. Bayliss^§, Sarah Howat, Catherine Davidson^¶, and Jayesh Dudhia

From the  Royal Veterinary College, Royal College Street, London NW1 0TU, United Kingdom and the ^¶ Wellcome/CRC Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, United Kingdom

The effect of age on the incorporation of newly synthesized aggrecan into the extracellular matrix of human articular cartilage was investigated. This property was measured in a pulse-chase explant culture system by determining the distribution of radiolabeled molecules ([³⁵S]sulfate-labeled) between a nondissociating extract (phosphate-buffered saline), which extracts mainly nonaggregated macromolecules, and a dissociating extract (4 M GnHCl) containing mainly aggrecan that was complexed in situ with hyaluronan. The rate of incorporation of aggrecan into aggregates was much slower in mature cartilage than in tissue obtained from younger individuals. Furthermore, autoradiography showed that in mature cartilage, newly synthesized aggrecan is not transported from the pericellular environment within the first 18 h of chase culture, whereas in immature cartilage, it moves into the intercellular space during the same period, i.e. aggrecan is processed in the extracellular space very differently in young and adult articular cartilage. Experiments were also performed to show that the interaction of link protein with newly synthesized aggrecan depends on the maturity of the G₁ domain of aggrecan. This investigation has shown that the extracellular aggregation of aggrecan in adult human articular cartilage involves a number of intermediate structures. These have not been identified in the very young cartilage obtained from laboratory animals or in porcine and bovine articular cartilage obtained from the abattoir.

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