The calcification of cartilage matrix in chondrocyte culture: studies of the C-propeptide of type II collagen (chondrocalcin).
We have shown that when chondrocytes are isolated by collagenase digestion of hyaline cartilage from growth plate, nasal, and epiphyseal cartilages of bovine fetuses they rapidly elaborate an extracellular matrix in culture. Only growth plate chondrocytes can calcify this matrix as ascertained by incorporation of 45Ca2+, detection of mineral with von Kossa's stain and electron microscopy. There is an extremely close direct correlation between 45Ca2+ incorporation in the first 24 h of culture and the content of the C-propeptide of type II collagen, measured by radioimmunoassay, at the time of isolation and during culture. Moreover, growth plate cells have an increased intracellular content of the C-propeptide per deoxyribonucleic acid and, during culture, per hydroxyproline (as a measure of helical collagen) compared with nasal and epiphyseal chondrocytes. In growth plate chondrocytes 24,25-dihydroxycholecalciferol (24,25-[OH]2D3), but not 1,25-dihydroxycholecalciferol alone, stimulates the net synthesis of the C-propeptide and calcification; proteoglycan net synthesis is unaffected. Together, these metabolites of vitamin D further stimulate C-propeptide net synthesis but do not further increase calcification stimulated by 24,25-(OH)2D3. These observations further demonstrate the close correlation between the C-propeptide of type II collagen and the calcification of cartilage matrix.
D015650:24,25-Dihydroxyvitamin D 3; D000818:Animals; D002113:Calcification, Physiologic; D002117:Calcitriol / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000032:analysis / Q000235:genetics; D002356:Cartilage / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D002417:Cattle; D002478:Cells, Cultured; D003094:Collagen / Q000032:analysis / Q000235:genetics; D024043:Collagen Type II; D004100:Dihydroxycholecalciferols / Q000494:pharmacology; D005333:Fetus; D006132:Growth Plate / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D006909:Hydroxyproline / Q000032:analysis; D008854:Microscopy, Electron
D002135:Calcium-Binding Proteins; D024043:Collagen Type II; D004100:Dihydroxycholecalciferols; C040072:chondrocalcin; D015650:24,25-Dihydroxyvitamin D 3; D003094:Collagen; D002117:Calcitriol; D006909:Hydroxyproline; D002118:Calcium