Collagen type I antisense and collagen type IIA messenger RNA is expressed in adult murine articular cartilage.
OBJECTIVE
Articular cartilage has only limited capacities for repair and it is not known what is the exact mechanism of matrix restoration. It was investigated whether the reparative process in murine articular cartilage after moderate proteoglycan depletion is accompanied by a change in the chondrocyte phenotype either to hypertrophy or to a less differentiated phenotype as assayed by the expression of specific collagen subtypes.


METHODS
Moderate proteoglycan depletion was induced by injection of papain whereafter the expression of collagen type I mRNA, collagen IIA and IIB mRNA and type X collagen mRNA in patellar cartilage, as markers for chondrocyte phenotype, was investigated by RT-PCR during normal cartilage physiology and matrix restoration. In addition, in-situ expression of collagen subtypes was assayed by immunolocalisation.


RESULTS
In normal articular cartilage collagen I, collagen IIB and collagen type X transcripts were easily detected. Surprisingly, collagen type I sense as well as antisense mRNA was detected and in addition to IIB transcripts collagen IIA transcripts were detected in a number of samples. During cartilage matrix restoration no change in the expression of collagen I, collagen IIA or IIB or collagen type X mRNA transcripts could be detected. Immunolocalization demonstrated the presence of type I (pericellular) and type II collagen in the extracellular matrix. The pericellular matrix of hypertrophic chondrocytes showed collagen type X staining in the calcified cartilage in normal and papain-injected knee joints. Increased staining for collagen type X was found in the upper cartilage layer in the interterritorial matrix from day 7 after papain injection.


CONCLUSIONS
The absence of changes in collagen mRNA expression indicates that alteration of chondrocyte phenotype does not occur during the successful repair process after moderate proteoglycan depletion. Collagen type X appears to be deposited in the upper cartilage layer during this process.
van der Kraan
P M
PM
Department of Rheumatology, University Hospital Nijmegen, The Netherlands. P.vanderKraan@reuma.azn.nl
Vitters
E L
EL
Meijers
T H
TH
Poole
A R
AR
van den Berg
W B
WB
D000818:Animals; D002358:Cartilage, Articular / Q000187:drug effects / Q000378:metabolism*; D003094:Collagen / Q000032:analysis / Q000235:genetics*; D015870:Gene Expression; D007150:Immunohistochemistry; D007719:Knee Joint; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D010206:Papain; D011509:Proteoglycans / Q000378:metabolism; D016372:RNA, Antisense / Q000032:analysis / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D011509:Proteoglycans; D016372:RNA, Antisense; D012333:RNA, Messenger; D003094:Collagen; D010206:Papain