Unloading results in rapid loss of TGFβ signaling in articular cartilage: role of loading-induced TGFβ signaling in maintenance of articular chondrocyte phenotype?
Recently it was shown that loading of articular cartilage explants activates TGFβ signaling. Here we investigated if in vivo chondrocytes express permanently high TGFβ signaling, and the consequence of the loss of compressive loading-mediated TGFβ signaling on chondrocyte function and phenotype.



Bovine articular cartilage explants were collected within 10 min post mortem and stained immediately and after 30, 60 and 360 min for phosphorylated-Smad2, indicating active TGFβ signaling. Explants were unloaded for 48 h and subsequently repeatedly loaded with a compressive load of 3 MPa. In addition, explants were cultured unloaded for 2 weeks and the effect of loading or exogenous TGFβ on proteoglycan level and chondrocyte phenotype (Col10a1 mRNA expression) was analyzed.



Unloading of articular cartilage results in rapid loss of TGFβ signaling while subsequent compressive loading swiftly restored this. Loading and exogenous TGFβ enhanced expression of TGFβ1 and ALK5. Unloading of explants for 2 weeks resulted in proteoglycan loss and increased Col10a1 expression. Both loading and exogenous TGFβ inhibited elevated Col10a1 expression but not proteoglycan loss.



Our data might imply that in vivo regular physiological loading of articular cartilage leads to enduring TGFβ signaling and TGFβ-induced gene expression. We propose a hypothetical model in which loading activates a self-perpetuating system that prevents hypertrophic differentiation of chondrocytes and is crucial for cartilage homeostasis.
Orthopedic Research Laboratory, Radboudumc, PO Box 9101, 6500 HB Nijmegen, The Netherlands.
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Madej
Experimental Rheumatology, Radboudumc, PO Box 9101, 6500 HB Nijmegen, The Netherlands.
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van Caam
Experimental Rheumatology, Radboudumc, PO Box 9101, 6500 HB Nijmegen, The Netherlands.
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Blaney Davidson
Orthopedic Research Laboratory, Radboudumc, PO Box 9101, 6500 HB Nijmegen, The Netherlands.
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Buma
Experimental Rheumatology, Radboudumc, PO Box 9101, 6500 HB Nijmegen, The Netherlands. Electronic address: peter.vanderkraan@radboudumc.nl.
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van der Kraan
D000818:Animals; D002358:Cartilage, Articular; D002417:Cattle; D019902:Chondrocytes; D010641:Phenotype; D011509:Proteoglycans; D016212:Transforming Growth Factor beta
D011509:Proteoglycans; D016212:Transforming Growth Factor beta
Articular cartilage; Hypertrophy; Loading; Smad; TGFβ