Subchondral and epiphyseal bone remodeling following surgical transection and noninvasive rupture of the anterior cruciate ligament as models of post-traumatic.

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Subchondral and epiphyseal bone remodeling following surgical transection and noninvasive rupture of the anterior cruciate ligament as models of post-traumatic osteoarthritis T. Maerz, M. Kurdziel, M.D. Newton, P. Altman, K. Anderson, H.W.T. Matthew, K.C. Baker Osteoarthritis and Cartilage Volume 24, Issue 4, Pages 698-708 (April 2016) DOI: 10.1016/j.joca.2015.11.005 Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Noninvasive ACL rupture was induced using the tibial compression model with which a rapid 3 mm displacement is applied to the tibia via the dorsiflexed paw, as previously characterized27. During rupture loading, the knee is flexed to ∼100° and held within a trough constraining only medial/lateral motion (A). Rupture was confirmed by an anterior drawer test, whereby anterior joint laxity is confirmed by the application of an anterior tibial force (B, C). Osteoarthritis and Cartilage 2016 24, 698-708DOI: (10.1016/j.joca.2015.11.005) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Segmentation process of femoral and tibial subchondral bone. Femoral and tibial articular cartilage was contrast enhanced via EPIC-μCT (A, E) to enable manual outlining (B, F, grey ROI) and segmentation of articular cartilage (B, F, black ROI) using a 3D region-growing algorithm. Subchondral bone was then segmented automatically (C, G, black ROI) using a dilation and thresholding algorithm to yield femoral (D) and tibial (H) subchondral bone volumes. Osteoarthritis and Cartilage 2016 24, 698-708DOI: (10.1016/j.joca.2015.11.005) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Segmentation process of femoral and tibial epiphyseal bone. Raw images of the femur (A) and tibia (D) were manually outlined to segment epiphyseal bone. Subchondral and cortical bone was removed from the volume of interest using voxel erosion (B, E, black ROI) to yield trabecular epiphyseal bone volumes (C, F). Osteoarthritis and Cartilage 2016 24, 698-708DOI: (10.1016/j.joca.2015.11.005) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Histology and μCT images of the medial compartment of representative Control (left column), Transection (center column), and Rupture (right column) animals at 4 weeks. In the femur, compared to Control animals (A, D), an overall loss of trabecular and subchondral bone volume is evident in both Rupture (B, E) and Transection (C, F), though more pronounced in Transection. Only Transection exhibited notable decreases in femoral (SCB.Th). In the tibia, compared to Control animals (G, J), only Transection (I, L) exhibited a marked decrease in trabecular bone volume, whereas both Rupture (H, K) and Transection (I, L) showed a decrease in subchondral bone volume. Osteoarthritis and Cartilage 2016 24, 698-708DOI: (10.1016/j.joca.2015.11.005) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Subchondral bone morphometric parameters in Control, Rupture, and Transection. Both Rupture and Transection exhibit loss of subchondral BV/TV in the femur (A) and tibia (B). Transection exhibited higher loss of femoral and tibial subchondral BV/TV (A, B). Rupture had significantly higher TMD in the femur (C) and tibia (D) compared to Transection at both 4 and 10 weeks. Femoral subchondral bone plate thickness (SCB.Th.Mean) was decreased significantly only in Transection (E) and there were significant differences in SCB.Th.Mean in all femoral compartments between Rupture and Transection at 4 weeks (E). Only subtle changes were observed in tibial SCB.Th.Mean (F). * indicates significant difference to Control. ♢ indicates significant difference between Rupture and Transection. Error bars represent 95% confidence interval. Osteoarthritis and Cartilage 2016 24, 698-708DOI: (10.1016/j.joca.2015.11.005) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 6 Epiphyseal bone BV/TV and TMD in Control, Rupture, and Transection. Only Transection exhibited significant decreases in epiphyseal BV/TV in both the femur (A) and tibia (B). Femoral epiphyseal TMD (C) was significantly lower in Transection at both 4 and 10 weeks compared to Rupture, and at 10 weeks, Rupture had higher femoral epiphyseal TMD compared to Control. No differences in tibial epiphyseal TMD were observed between groups at either time point (D). * indicates significant difference to Control. ♢ indicates significant difference between Rupture and Transection. Error bars represent 95% confidence interval. Osteoarthritis and Cartilage 2016 24, 698-708DOI: (10.1016/j.joca.2015.11.005) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 7 Epiphyseal bone Tb.Th.Mean, Tb.N, and Tb.Sp.Mean in Control, Rupture, and Transection. Rupture had significantly higher femoral Tb.Th.Mean compared to Control and Transection at 4 weeks, and both injury groups exhibited higher Tb.Th.Mean compared to Control at 10 weeks (A). No significant differences in tibial Tb.Th.Mean were observed between groups (B). Compared to Control, Tb.N was significantly decreased in the femur (C) and tibia (D) of both injury groups at both time points. Neither injury group exhibited significant changes in Tb.Sp.Mean in either the femur (E) or tibia (F). * indicates significant difference to Control. ♢ indicates significant difference between Rupture and Transection. Error bars represent 95% confidence interval. Osteoarthritis and Cartilage 2016 24, 698-708DOI: (10.1016/j.joca.2015.11.005) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 8 Epiphyseal bone SMI and Conn.D in Control, Rupture, and Transection. Both Rupture and Transection exhibited increases in femoral (A) and tibial (B) SMI at 4 and 10 weeks. No significant changes in Conn.D were observed compared to Control in either injury group in the femur (C) or tibia (D), but Transection exhibited significantly higher Conn.D compared to Rupture in the femur at 4 weeks (C). * indicates significant difference to Control. ♢ indicates significant difference between Rupture and Transection. Error bars represent 95% confidence interval. Osteoarthritis and Cartilage 2016 24, 698-708DOI: (10.1016/j.joca.2015.11.005) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions