Type X Collagen Degradation in Long-Term Serum-Free Culture of the Embryonic Chick Tibia Following Production of Active Collagenase and Gelatinase

Doxycycline disrupts chondrocyte differentiation and inhibits cartilage matrix degradation

“…Chicken collagenase activity has previously been shown to be associated with cleared bands at 57 kd and 50 kd (10). Gelatinase appeared as a doublet of bands at 66 kd and 62 kd.…”

“…The presence of gelatinolytic proteinases in the CM was assayed by gelatinsubstrate zymography using a modification of the method of Heussen and Dowdle (16), as previously described (10). Gelatin was incorporated in 8% gels to a final concentration of 0.5 mg/ml.…”

“…Collagen release into the CM was measured as hydroxyproline content. Total hydroxyproline content of the uncultured tibias was determined by a modified Steggman procedure (17), as previously described (10). Purified rat tail type I collagen was used as a standard, where 1 pg hydroxyproline is equivalent to 8.5 pg of collagen.…”

“…Chondroitin sulfate was used as standard. GAG content was expressed as pg GAG/130 pl of CM/tibia, as previously described (10).…”

Gelatinase-mediated migration and invasion of cancer cells

“…Indeed, some substrates show over 200-fold selectivity towards MMP-2 Kridel et al, 2001). (Morodomi et al, 1992;Okada et al, 1990) Native collagen type: I yes no (Aimes and Quigley, 1995) III yes no (Berton et al, 2000) IV yes yes (Morodomi et al, 1992) V yes yes (Morodomi et al, 1992) VII yes (Seltzer et al, 1989) X yes (Cole et al, 1993) XI yes (Smith et al, 1991) XVIII yes (Ferreras et al, 2000) Aggrecan yes yes (Fosang et al, 1992) BM-40/SPARC/ Osteonectin yes yes (Sasaki et al, 1997) Brevican yes no Decorin yes no (Imai et al, 1997) Elastin yes yes Entactin/nidogen no yes (Mayer et al, 1993;Sires et al, 1993) Fibrillin yes yes (Ashworth et al, 1999) Fibrin yes (Lelongt et al, 2001) Fibrinogen yes yes (Bini et al, 1996;Lelongt et al, 2001) Fibronectin yes no (Okada et al, 1990) Laminin yes yes (Giannelli et al, 1997;Morodomi et al, 1992) Link protein yes yes (Nguyen et al, 1993) NG2 proteoglycan yes (Larsen et al, 2003) Neurocan yes (Turk et al, 2001) Tenascin yes no (Siri et al, 1995) Vitronectin yes yes (Imai et al, 1995) α2-macroglobulin yes yes (Arbelaez et al, 1997) αB-crystallin yes (Starckx et al, 2003) Amyloid protein precursor yes (LePage et al, 1995) Big endothelin-1 yes yes (Fernandez-Patron et al, 2002) Calcitonin gene-related peptide (CGRP) yes (Fernandez-Patron et al, 2000) Complement protein C1q yes yes (Ruiz et al, 1999) Connective tissue-activating peptide-III (CTAP-III) yes (Van den Eph B1 tyrosine kinase receptor yes no Epithelial-cell derived neutrophil activating pept...…”