ADAMTS5−/− mice have less subchondral bone changes after induction of osteoarthritis through surgical instability: implications for a link between cartilage.

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ADAMTS5−/− mice have less subchondral bone changes after induction of osteoarthritis through surgical instability: implications for a link between cartilage and subchondral bone changes S.M. Botter, S.S. Glasson, B. Hopkins, S. Clockaerts, H. Weinans, J.P.T.M. van Leeuwen, G.J.V.M. van Osch Osteoarthritis and Cartilage Volume 17, Issue 5, Pages 636-645 (May 2009) DOI: 10.1016/j.joca.2008.09.018 Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Selection procedure of tibial epiphysis for 3D-morphometric analysis. The epiphysis was manually selected as representative of subchondral bone, and outgrowing osteophytes were excluded. Epiphyseal trabeculae were separated from the subchondral bone plate and trabecular bone morphometric parameters were calculated (see Table I). A subset of the medial and lateral subchondral bone plate and underlying epiphyseal trabecular bone were analyzed separately, as indicated for the medial side. Osteoarthritis and Cartilage 2009 17, 636-645DOI: (10.1016/j.joca.2008.09.018) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 (A) OA scores summed over the four analyzed quadrants (medial tibial plateau, medial femoral condyle, lateral tibial plateau, and lateral femoral condyle) from left unoperated control (Ctrl) knee joints and right knee joints that underwent dissection of the medial meniscus (DMM) of WT and ADAMTS5−/− animals at 18 weeks of age. Microphotographs on the right show examples of the amount of cartilage damage found in DMM joints of WT and ADAMTS5−/− mice (original magnification 200×). (B) OA scores of the medial and lateral tibial plateau separately. Shown are averages±s.e.m., P<0.05 was considered significant. Osteoarthritis and Cartilage 2009 17, 636-645DOI: (10.1016/j.joca.2008.09.018) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 (A) Representative cross-sections of tibiae from WT and ADAMTS5−/− mice, both left unoperated control (Ctrl) and right DMM. The inset shows a 3D reconstruction of a proximal tibia with a plane indicating the location from which the cross-sections were taken. Epiphysis (E), metaphysis (M) and growth plate (dotted line) are indicated, as well as the medial (Med) and lateral (Lat) tibial plateau. Subchondral sclerosis was found in the medial region of the epiphysis (arrows), and medial osteophytes were observed as well (arrowheads). (B) Histological images of the medial tibia from DMM WT and ADAMTS5−/− mice, stained with toluidine-blue. Please note the pronounced sclerosis of the subchondral bone plate in the WT mice, whereas ADAMTS5−/− mice only showed mild sclerosis. Magnification 100×. C=cartilage, O=osteophyte, G=growth plate. Osteoarthritis and Cartilage 2009 17, 636-645DOI: (10.1016/j.joca.2008.09.018) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 (A) Subchondral bone plate thickness (Sb. plate Th.) measured with micro-CT in left unoperated control (Ctrl) and right DMM knee joints, in WT and ADAMTS5−/− mice. The percental increases in plate thickness in DMM joints as compared to the control joints are indicated inside the DMM bars. (B) Subchondral bone volume fraction (Sb BV/TV). (C) Calcified cartilage thickness, measured from histological images. Shown are averages±s.e.m., P<0.05 was considered significant. Osteoarthritis and Cartilage 2009 17, 636-645DOI: (10.1016/j.joca.2008.09.018) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Examples of TRAP-stained osteoclasts in the medial tibia of WT and ADAMTS5−/− mice, both left unoperated control (Ctrl) and right DMM. Osteoclasts in the subchondral bone (black arrowheads) were observed in Ctrl and DMM joints of both genotypes, in addition to osteoclasts beneath the growth plate (white arrowheads). Please note that in DMM joints of ADAMTS5−/− mice, very few if any osteoclasts were observed despite the presence of osteoclasts beneath the growth plate. Also note the thinner subchondral bone plate in the ADAMTS5−/− joints and the subchondral sclerosis after DMM in the WT joint. Magnification 100×. C=cartilage, SBP=subchondral bone plate, G=growth plate. Osteoarthritis and Cartilage 2009 17, 636-645DOI: (10.1016/j.joca.2008.09.018) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions