Proteolysis involving matrix metalloproteinase 13 (collagenase-3) is required for chondrocyte differentiation that is associated with matrix mineralization.
Collagenases are involved in cartilage matrix resorption. Using bovine fetal chondrocytes isolated from physeal cartilages and separated into a distinct prehypertrophic subpopulation, we show that in serum-free culture they elaborate an extracellular matrix and differentiate into hypertrophic chondrocytes. This is characterized by expression of type X collagen and the transcription factor Cbfal and increased incorporation of 45Ca2+ in the extracellular matrix, which is associated with matrix calcification. Collagenase activity, attributable only to matrix metalloproteinase (MMP) 13 (collagenase-3), is up-regulated on differentiation. A nontoxic carboxylate inhibitor of MMP-13 prevents this differentiation; it suppresses expression of type X collagen, Cbfal, and MMP-13 and inhibits increased calcium incorporation in addition to inhibiting degradation of type II collagen in the extracellular matrix. General synthesis of matrix proteins is unaffected. These results suggest that proteolysis involving MMP-13 is required for chondrocyte differentiation that occurs as part of growth plate development and which is associated with matrix mineralization.
Wu
C William
CW
Joint Diseases Laboratory, Shriners Hospitals for Children, Canadian Hospital, Montreal, Quebec.
Tchetina
Elena V
EV
Mwale
Fackson
F
Hasty
Karen
K
Pidoux
Isabelle
I
Reiner
Agnes
A
Chen
Jeffrey
J
Van Wart
Harold E
HE
Poole
A Robin
AR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism; D002264:Carboxylic Acids / Q000494:pharmacology; D002417:Cattle; D002454:Cell Differentiation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology*; D003001:Cloning, Molecular; D024043:Collagen Type II / Q000378:metabolism; D017364:Collagenases / Q000235:genetics* / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005109:Extracellular Matrix / Q000378:metabolism*; D005347:Fibroblasts / Q000201:enzymology; D006132:Growth Plate / Q000166:cytology / Q000201:enzymology; D015576:Hyperostosis / Q000201:enzymology / Q000473:pathology; D007211:Indoles / Q000494:pharmacology; D020781:Matrix Metalloproteinase 1 / Q000235:genetics; D053509:Matrix Metalloproteinase 13; D061965:Matrix Metalloproteinase Inhibitors; D008903:Minerals / Q000378:metabolism; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins; D014157:Transcription Factors / Q000187:drug effects / Q000378:metabolism
D002264:Carboxylic Acids; D024043:Collagen Type II; D004791:Enzyme Inhibitors; D007211:Indoles; D061965:Matrix Metalloproteinase Inhibitors; D008903:Minerals; D009363:Neoplasm Proteins; C406074:RS 102481; D014157:Transcription Factors; D017364:Collagenases; D053509:Matrix Metalloproteinase 13; D020781:Matrix Metalloproteinase 1; D002118:Calcium