Juha Töyräs

Comparison of the equilibrium response of articular cartilage in unconfined compression, confined compression and indentation

At mechanical equilibrium, articular cartilage is usually characterized as an isotropic elastic material with no interstitial fluid flow. In this study, the equilibrium properties (Youngs modulus, aggregate modulus and Poissons ratio) of bovine humeral, patellar and femoral cartilage specimens (n=26) were investigated using unconfined compression, confined compression, and indentation tests. Optical measurements of the Poissons ratio of cartilage were also carried out. Mean values of the Youngs modulus (assessed from the unconfined compression test) were 0.80+/-0.33, 0.57+/-0.17 and 0.31+/-0.18MPa and of the Poissons ratio (assessed from the optical test) 0.15+/-0.06, 0.16+/-0.05 and 0.21+/-0.05 for humeral, patellar, and femoral cartilages, respectively. The indentation tests showed 30-79% (p<0.01) higher Youngs modulus values than the unconfined compression tests. In indentation, values of the Youngs modulus were independent of the indenter diameter only in the humeral cartilage. The mean values of the Poissons ratio, obtained indirectly using the mathematical relation between the Youngs modulus and the aggregate modulus in isotropic material, were 0.16+/-0.06, 0.21+/-0.05, and 0.26+/-0.08 for humeral, patellar, and femoral cartilages, respectively. We conclude that the values of the elastic parameters of the cartilage are dependent on the measurement technique in use. Based on the similar values of Poissons ratios, as determined directly or indirectly, the equilibrium response of articular cartilage under unconfined and confined compression is satisfactorily described by the isotropic elastic model. However, values of the isotropic Youngs modulus obtained from the in situ indentation tests are higher than those obtained from the in vitro unconfined or confined compression tests and may depend on the indenter size in use.

Novel mechano-acoustic technique and instrument for diagnosis of cartilage degeneration

Fibrillation of articular surface and depletion of proteoglycans are the structural changes related to early osteoarthrosis. These changes make cartilage softer and prone to further degeneration. The aim of the present study was to combine mechanical and acoustic measurements towards quantitative arthroscopic evaluation of cartilage quality. The performance of the novel ultrasound indentation instrument was tested with elastomers and bovine articular cartilage in vitro. The instrument was capable of measuring elastomer thickness (r = 1.000, p < 0.01, n = 8) and dynamic modulus (r = 0.994, p < 0.01, n = 13) reliably. Osteochondral plugs were tested before and after enzymatic degradation of cartilage proteoglycans by trypsin or chondroitinase ABC, and of cartilage collagens by collagenase. Trypsin and collagenase induced a mean decrease of -31.2 +/- 12.3% (+/- SD, p < 0.05) and -22.9 +/- 20.8% (p = 0.08) in dynamic modulus, respectively. Rate of cartilage deformation, i.e. creep rate, increased by +117.8 +/- 71.4% (p < 0.05) and +24.7 +/- 35.1% (p = 0.17) in trypsin and chondroitinase ABC treatments, respectively. Collagenase induced a greater decrease in the ultrasound reflection from the cartilage surface (-54.2 +/- 29.6%, p < 0.05) than trypsin (-17.1 +/- 13.5%, p = 0.08). In conclusion, combined quantitation of tissue modulus, viscoelasticity and ultrasound reflection from the cartilage surface provides a sensitive method to distinguish between normal and degenerated cartilage, and even to discern proteoglycan loss and collagen degradation from each other.

REAL-TIME ULTRASOUND ANALYSIS OF ARTICULAR CARTILAGE DEGRADATION IN VITRO

The sensitivity of the reflection coefficient, attenuation and velocity to the enzymatic degradation of bovine patellar cartilage was evaluated in real-time with high-frequency ultrasound (US) (29.4 MHz). These parameters were estimated from the radiofrequency (RF) signal, which was recorded at 5-min intervals during the digestion of the tissue by collagenase or by trypsin. The coefficient of reflection at cartilage surface decreased by 78.5% and 10.5% (p < 0.05) after 6 h of exposure to collagenase and 4 h of exposure to trypsin, respectively. During the trypsin digestion, the attenuation in cartilage increased by 0.274 dB/mm (p < 0.05) and the velocity decreased by 7 m/s (p < 0.05). The coefficient of reflection at the cartilage surface was the most sensitive acoustic parameter to the enzymatic degradation of cartilage and may be the easiest to implement for clinical diagnosis of cartilage quality. US velocity was found to be insensitive to degradation. The small difference in mean velocity between the control and degraded cartilage suggests that a constant predefined US velocity value can be used to obtain diagnostically acceptable measurement of the cartilage thickness.

Ultrasound indentation of bovine knee articular cartilage in situ

We have earlier developed a handheld ultrasound indentation instrument for the diagnosis of articular cartilage degeneration. In ultrasound indentation, cartilage is compressed with the ultrasound transducer. Tissue thickness and deformation are calculated from the A-mode ultrasound signal and the stress applied is registered with the strain gauges. In this study, the applicability of the ultrasound indentation instrument to quantify site-dependent variation in the mechano-acoustic properties of bovine knee cartilage was investigated. Osteochondral blocks (n=6 per site) were prepared from the femoral medial condyle (FMC), the lateral facet of the patello-femoral groove (LPG) and the medial tibial plateau (MTP). Cartilage stiffness (dynamic modulus, E(dyn)), as obtained with the ultrasound indentation instrument in situ, correlated highly linearly (r=0.913, p<0.01) with the values obtained using the reference material-testing device in vitro. Reproducibility (standardized coefficient of variation) of the ultrasound indentation measurements was 5.2%, 1.7% and 3.1% for E(dyn), ultrasound reflection coefficient of articular surface (R) and thickness, respectively. E(dyn) and R were site dependent (p<0.05, Kruskall-Wallis H test). E(dyn) was significantly higher (p<0.05, Kruskall-Wallis Post Hoc test) in LPG (mean+/-SD: 10.1+/-3.1MPa) than in MTP (2.9+/-1.4MPa). In FMC, E(dyn) was 4.6+/-1.3MPa. R was significantly (p<0.05) lower at MTP (2.0+/-0.7%) than at other sites (FMC: 4.2+/-0.9%; LPG: 4.4+/-0.8%). Cartilage glycosaminoglycan concentration, as quantified with the digital densitometry, correlated positively with E(dyn) (r=0.678, p<0.01) and especially with the equilibrium Youngs modulus (reference device, r=0.874, p<0.01) but it was not associated with R (r=0.294, p=0.24). We conclude that manual measurements are reproducible and the instrument may be used for detection of cartilage quality in situ. Especially, combined measurement of thickness, E(dyn) and R provides valuable diagnostic information on cartilage status.

Mechano-acoustic diagnosis of cartilage degeneration and repair.

Background: The combined use of high-frequency ultrasound and mechanical indentation has been suggested for the evaluation of cartilage integrity. In this study, we investigated the usefulness of high-resolution B-mode ultrasound imaging and quantitative mechanical measurements for the diagnosis of cartilage degeneration and for monitoring tissue-healing after autologous chondrocyte transplantation.Methods: In the first study, osteochondral samples (n = 32) were obtained from the lateral facet of a bovine patella, and the samples were visually classified as intact (n = 13) or degenerated (n = 19) and were graded with use of the Mankin scoring system. Samples were imaged with use of a 20-MHz ultrasound instrument, and the dynamic modulus (Edyn) of cartilage was determined in unconfined compression with use of a high-resolution materials tester. In the second study, cartilage chondrocytes were harvested from the low-weight-bearing area of six-month-old porcine knee joints and cultured. A month later, a cartilage lesion was created on the facet of the femoral trochlea and was repaired with use of the autologous chondrocyte transplantation technique (n = 10). Three months later, to estimate cartilage Edyn, the repair tissue, the adjacent cartilage, and the sham-operated contralateral joint cartilage (control) were analyzed in situ with an arthroscopic indentation instrument. Subsequently, the same sites were imaged with ultrasound.Results: All visually degenerated bovine samples (mean Mankin score = 4) and five visually normal samples (Mankin score = 1) showed reduced Edyn (<2.1 MPa) as compared with histologically normal cartilage (Edyn = 13.8 ± 3.2 MPa, Mankin score = 0). Cartilage stiffness, as shown by the indenter force, was lower (0.6 ± 0.3 N, p < 0.05, Wilcoxons signed-rank test) in the porcine tissue repaired with autologous chondrocyte transplantation than it was in the adjacent (1.6 ± 0.1 N) or the control (1.9 ± 0.4 N) tissue. The superficial and internal structure of the degenerated and repaired tissue, including the subchondral erosion at the repair site, was sensitively demonstrated by the ultrasound imaging.Conclusions: Measurement of cartilage Edyn is an objective method with which to follow changes in the mechanical integrity of cartilage. B-mode ultrasound imaging offers detailed information on the structural properties of cartilage and subchondral bone.Clinical Relevance: Mechanical indentation and ultrasound imaging complement each other and provide information on the functional and structural integrity of cartilage and subchondral bone. Combined use of these techniques may provide a means for the early diagnosis of cartilage degeneration and for the monitoring of tissue healing after repair surgery.