This study examines the basic metabolic events of aggrecan synthesis and breakdown in the growth plate at different depths and at different stages of development. Growth plate was harvested from the distal tibia of fetal and calf tissue and maintained as explants in serum-free-conditions. The tissue was sectioned into three equal depths (resting/proliferative zone, upper hypertrophic zone, and lower hypertrophic zone) and (a) cultured for three days with daily media change for studies of prteoglycan breakdown rates, or (b) incubated with [35S]-sulfate to determine relative rates of proteoglycan synthesis. Rates of both aggrecan synthesis and turnover were highest in the resting/proliferative zone compared to the upper or lower hypertrophic zones, and was greater in the calf compared to the fetal tissue. In situ hybridization studies showed that aggrecan gene expression in the cells of the resting/proliferative zone and the upper hypertrophic zones were similar, and was reduced in the deepest cells of the lower hypertrophic zone, adjacent to the zone of calcification. Proteoglycan structure was characterized associative and dissociative Sepharose CI2B chromatography. These results showed that approximately 90% of the newly synthesized proteoglycan, and the total proteoglycan population, was able to aggregate and that the monomers were relatively large. The proteoglycan released into the media had a reduced ability to aggregate and the monomers were of a more variable size. These data support the hypothesis that the matrix proteoglycan content is controlled both by the rate of synthesis and breakdown, but in the lower regions the rate of synthesis may play a more dominant role. The higher metabolic activity of aggrecan in the calf than fetal growth plate may be a result of environmental stimuli (i.e., soluble mediators, loading) during different stages of development.
- growth plate