Objective The aim of this study is to evaluate whether contrast enhanced computed tomography (CECT) attenuation, using a cationic contrast agent (CA4+), correlates with the equilibrium compressive modulus (E) and coefficient of friction (μ) of ex vivo bovine articular cartilage. Methods Correlations between CECT attenuation and E (Group 1, n=12) and μ (Group 2, n=10) were determined using 7mm diameter bovine osteochondral plugs from the stifle joints of six freshly slaughtered, skeletally mature cows. The equilibrium compressive modulus was measured using a 4-step, unconfined, compressive stress relaxation test, and the coefficients of friction were determined from a torsional friction test. Following mechanical testing, samples were immersed in CA4+, imaged using μCT, rinsed, and analyzed for glycosaminoglycan (GAG) content using the DMMB assay. Results The CECT attenuation was positively correlated with the GAG content of bovine cartilage (R2= 0.87, p<0.0001 for Group 1 and R2= 0.74, p=0.001 for Group 2). Strong and significant positive correlations were observed between E and GAG content (R2= 0.90, p<0.0001) as well as CECT attenuation and E (R2= 0.90, p<0.0001). The CECT attenuation was negatively correlated with the three coefficients of friction: CECT vs. μstatic (R2=0.71, p=0.002), CECT vs. μstatic_equilibrium (R2=0.79, p<0.001), and CECT vs. μkinetic (R2=0.69, p=0.003). Conclusions CECT with CA4+ is a useful tool for determining the mechanical properties of ex vivo cartilage tissue as the attenuation significantly correlates with the compressive modulus and coefficient of friction.
Mouse models of osteoarthritis (OA) are commonly used to study the disease’s pathogenesis and efficacy of potential treatments. However, measuring the biochemical and mechanical properties of articular cartilage in these models currently requires destructive and time-consuming histology and mechanical testing. Therefore, we examined the feasibility of using contrast-enhanced CT (CECT) to rapidly and non-destructively image and assess the glycosaminoglycan (GAG) content. Using three ex vivo C57BL/6 mouse tibial plateaus, we determined the time required for the cationic contrast agent CA4+ to equilibrate in the cartilage. The whole-joint coefficient of friction (μ) of thirteen mouse knees (some digested with Chondroitenase ABC to introduce variation in GAG) was evaluated using a modified Stanton pendulum. For both the medial and lateral tibial plateau cartilage of these knees, linear regression was used to compare the equilibrium CECT attenuations to μ, as well as each side’s indentation equilibrium modulus (E) and Safranin-O determined GAG content. CA4+ equilibrated in the cartilage in 30.9 ± 0.95 min (mean ± SD, tau value of 6.17 ± 0.19 min). The mean medial and lateral CECT attenuation was correlated with μ (R2=0.69, p<0.05), and the individual medial and lateral CECT attenuations correlated with their respective GAG contents (R2≥0.63, p<0.05) and E (R2≥0.63, p<0.05). In conclusion, CECT using CA4+ is a simple, non-destructive technique for three-dimensional imaging of ex vivo mouse cartilage, and significant correlations between CECT attenuation and GAG, E, and μ are observed.
We determined whether contrast-enhanced computed tomography (CECT) attenuation obtained using a μCT scanner correlated with the glycosaminoglycan (GAG) content and distribution in ex vivo bovine menisci. Bovine samples were immersed in different concentrations of the contrast agents CA4+ and Ioxaglate, and the μCT images were compared to Safranin-O staining. CA4+ and Ioxaglate diffusion-in kinetics and the correlation between their CECT attenuations and GAG content were investigated. CA4+ and Ioxaglate both reached steady state in the meniscal regions within 95 h, with tau values of 20.6 ± 3.98 and 25.9 ± 3.71 h (mean ± SD), respectively. Both agents diffused preferentially through the proximal and secondarily through the distal surface. The CA4+ CECT attenuation was strongly and positively correlated with the GAG content of the meniscus regions (R2 = 0.89, p < 0.001) at low concentrations (12 mgI/ml), while the Ioxaglate CECT attenuation was moderately and negatively correlated with the GAG content (R2 = 0.51, p = 0.03) at 60 mgI/ml. CECT can image ex vivo menisci, and the CA4+, compared to Ioxaglate, enhanced attenuation strongly correlates with the GAG content and distribution in bovine meniscus.
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