Utilization of longitudinal ultrasound to quantify joint soft-tissue changes in a mouse model of posttraumatic osteoarthritis

Hao Xu, Echoe M Bouta, Ronald W Wood, Edward M Schwarz, Yongjun Wang, Lianping Xing, Hao Xu, Echoe M Bouta, Ronald W Wood, Edward M Schwarz, Yongjun Wang, Lianping Xing

Abstract

To assess the utility of longitudinal ultrasound (US) to quantify volumetric changes in joint soft tissues during the progression of posttraumatic osteoarthritis (PTOA) in mice, and validate the US results with histological findings. A longitudinal cohort of 3-month-old wild-type C57BL/6 male mice received the Hulth-Telhag surgical procedure on right knee to induce PTOA, and sham surgery on their left knee as control. US scans were performed on both knees before, 2, 4, 6, and 8 weeks post-surgery. Joint space volume and Power-Doppler (PD) volume were obtained from US images via Amira software. A parallel cross-sectional cohort of mice was killed at each US time point, and knee joints were subjected to histological analysis to obtain synovial soft-tissue area and OARSI scores. The correlation between US joint space volume and histological synovial soft-tissue area or OARSI score was assessed via linear regression analysis. US images indicated increased joint space volume in PTOA joints over time, which was associated with synovial inflammation and cartilage damage by histology. These changes started from 2 weeks post-surgery and gradually became more severe. No change was detected in sham joints. Increased joint space volume was significantly correlated with increased synovial soft-tissue area and the OARSI score (P<0.001). PD signal was detected in the joint space of PTOA joints at 6 weeks post-surgery, which was consistent with the location of blood vessels that stained positively for CD31 and alpha-smooth muscle actin in the synovium. This study indicates that US is a cost-effective longitudinal outcome measure of volumetric and vascular changes in joint soft tissues during PTOA progression in mice, which positively correlates with synovial inflammation and cartilage damage.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Orientation of joint space on an ultrasound image. A 3-month-old wild-type C57BL/6 male mouse was used. (a) An ultrasound B-mode image of a mouse knee joint. (b) Outlined joint space (solid green), tibia, and femur on the ultrasound B-mode image. (c) Alcian blue/Orange G (ABOG)-stained knee section with outlined joint space detected by ultrasound. (d) Illustration shows 3D reconstruction of join space derived from a stack of ultrasound B-mode images.
Figure 2
Figure 2
Joint space volume detected by ultrasound is positively correlated to change in synovial soft-tissue area in osteoarthritis (OA) joints. (a) Ultrasound B-mode images of joint space (red line) at before and different time points post-OA surgery. (b) 3D reconstruction from B-mode images. (c) ABHO-stained knee sections from sham and OA joints at different time points post surgery. Soft tissues in the area detected by US detected are outlined and analyzed. Changes of joint space volumes in OA (d) or sham joints (e) from 3D ultrasound. (f) The percentage of soft-tissue area (black line) over the total tissue area (blue line). (g) Correlation analysis of joint space volume and soft-tissue area (%). Values are from individual joint. n=8 mice/group. *P<0.05 between two indicated groups. w, weeks.
Figure 3
Figure 3
Joint space volume detected by ultrasound is positively correlated to cartilage damage in osteoarthritis (OA) joints. (a) ABHO-stained knee sections from sham and OA joints at different time points post surgery. (b) OARSI score from ABHO-stained sections. (c) Correlation analysis of joint space volume and OARSI score. Values are from individual joint. n=8 mice per group. *P<0.05 between the two indicated groups. w, weeks.
Figure 4
Figure 4
Power Doppler ultrasound detects blood vessels in later stages of osteoarthritis (OA) joints. (a) 3D reconstruction of blood vessels within the joint space in 8 weeks post-OA surgery. (b) H&E-stained knee joint section. (c) Enlarged image of two blood vessels. (d) Immunofluorescent staining of blood vessels with anti-CD31 antibody for endothelial cells (red) and anti-alpha-smooth muscle actin antibody for smooth muscle (green). Power Doppler volume in the joint space of OA (e) or sham joints (f) at different time points post-OA surgery. Values are from individual joint. n=8 mice per group. *P<0.05 between the two indicated groups. w, weeks.

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