Figure 5

Relaxin-induced collagenase activity contributes to loss of disc collagen. Conditioned medium from disc incubated with control medium (Ct), β-estradiol (Es), relaxin (R), or β-estradiol plus relaxin (Es+R) in the presence of the matrix metalloproteinase inhibitor GM6001 or its control analog was subjected to fluorescein isothiocyanate-labelled collagen degradation assay. The collagenase activity (relative fluorescence units [RFU]/ml) was standardized by the dry weight of the tissue (mg), and fold changes (means ± SD) were plotted (a, b). Disc digests from these experiments were assayed for collagen with the Sircol assay, and the results were standardized to tissue dry weight (mg). Fold changes in collagen concentration (means ± SD) were calculated and plotted (c, d). The untreated control (Ct) discs used in all experiments were exposed to control analog only. ** P < 0.01, *** P < 0.0001 by Fisher's test.