Effects on Matrix Degradation and Maintenance in Cartilage Explants

Effects on Matrix Degradation and Maintenance in Cartilage Explants 

A sterile stock solution of 5 mg/ml ascorbic acid in Ham s F12 is prepared, and aliquots are stored at -20 °C. 

Human recombinant interleukin-1 (IL-1) a or p from Genzyme is stored in aliquots at -20 °C. 

Fetlocks of freshly slaughtered steers (age 18 to 20 months) are skinned and the metacarpo-phalangeal joint opened under semi-sterile conditions. With a sterile punch (as used for obtaining skin biopsies) full thickness disks of cartilage are obtained from all accessible cartilaginous areas and their wet weight is assessed. In each well, 1 ml of medium is added, consisting of Ham s F12 supplemented with 10 % FCS 

Counts per minute (cpm) from medium and explant fractions are calculated related to mg wet weight of cartilage of the respective wells. They are either added if total incorporation is measured, or left separately, in case the ratio between released versus retained matrix is of interest. The data are converted into percent incorporation in comparison with the values of the untreated control or of the IL-1 control group serving as 100%. 

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I.L.1.2 Effects on Matrix Degradation and Maintenance in Cartilage Explants 245... 

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