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Cartilage viscoelasticity changes as cartilage degenerates. Hence, a cartilage viscoelasticity measurement could be an alternative to traditional imaging methods for osteoarthritis diagnosis. In a previous study, we confirmed the feasibility of viscoelasticity measurement in ex vivo bovine cartilage using the Lamb wave method. However, the wave speed-frequency curve of Lamb wave is totally nonlinear and the cartilage thickness could significantly affect the Lamb wave speed, making wave speed measurements and viscoelasticity inversion difficult. The objective of this study was to measure the cartilage viscoelasticity using the Rayleigh wave method (RWM). Rayleigh wave speed in the ex vivo bovine cartilage was measured, and exists only in the near-source and far-field region. The estimated cartilage elasticity was 0.66 ± 0.05 and 0.59 ± 0.07 MPa for samples 1 and 2, respectively; the estimated cartilage viscosity was 24.2 ± 0.7 and 27.1 ± 1.8 Pa·s for samples 1 and 2, respectively. These results were found to be highly reproducible, validating the feasibility of viscoelasticity measurement in ex vivo cartilage using the RWM. Current method of cartilage viscoelasticity measurement might be translated into in vivo application.

This study investigated wear damage of immature bovine articular cartilage using reciprocal sliding of tibial cartilage strips against glass or cartilage. Experiments were conducted in physiological buffered saline (PBS) or mature bovine synovial fluid (SF). A total of 63 samples were tested, of which 47 exhibited wear damage due to delamination of the cartilage surface initiated in the middle zone, with no evidence of abrasive wear. There was no difference between the friction coefficient of damaged and undamaged samples, showing that delamination wear occurs even when friction remains low under a migrating contact area configuration. No difference was observed in the onset of damage or in the friction coefficient between samples tested in PBS or SF. The onset of damage occurred earlier when testing cartilage against glass versus cartilage against cartilage, supporting the hypothesis that delamination occurs due to fatigue failure of the collagen in the middle zone, since stiffer glass produces higher strains and tensile stresses under comparable loads. The findings of this study are novel because they establish that delamination of the articular surface, starting in the middle zone, may represent a primary mechanism of failure. Based on preliminary data, it is reasonable to hypothesize that delamination wear via subsurface fatigue failure is similarly the primary mechanism of human cartilage wear under normal loading conditions, albeit requiring far more cycles of loading than in immature bovine cartilage.

Unique biomechanical behavior of articular cartilage is a result of its structure and composition. Interrelationships of tissue constituents (collagen, proteoglycans (PGs) and water) and tissue biomechanical parameters have been studied, but it is evident that no constituent alone explains the tissue mechanics. Fourier transform infrared (FT-IR) spectra can provide detailed information about the biochemical composition of articular cartilage. In this study, a chemometric approach to predict the biomechanical behavior of articular cartilage directly from the FT-IR spectra, i.e., without converting the data into collagen and PG information, was investigated. Partial least squares regression (PLSR) was used to predict equilibrium modulus (n=32) and dynamic modulus (n=24) of bovine cartilage samples from their average FT-IR spectra. The linear correlation coefficients between the reference and predicted values of Young's modulus and dynamic modulus were r=0.866 (p

We were the first to examine the mechanical responses of skeletally mature bovine femoral cartilage under large-strain simple shear (up to ±20%) using a multiaxial shear testing device. Since shear loading is critical in both tissue failure and chondrocyte responses, we aimed to probe (1) anisotropy with respect to the split-line direction (principal alignment of the collagen fibers near the articulating surface), (2) heterogeneity between femoral condyles, and (3) the influence of local cartilage thickness. We harvested a total of 48 cuboid cartilage specimens from four bovine knee joints. With each specimen we applied shear strains both parallel and perpendicular to the local split-line direction at a rate of 75μm/min and calculated the peak-to-peak shear stresses, shear strain-energy dissipation densities, and peak effective shear moduli. The Wilcoxon signed rank test revealed that the medial condyle was anisotropic in some mechanical measures at applied shear strains above 5%, while the lateral condyle was mechanically isotropic at all applied shear strains. The Kruskal-Wallis test revealed no significant differences in the median mechanical behavior of the lateral and medial condyles. Spearman׳s rank correlations revealed statistically significant negative monotonic correlations among thickness and most of our mechanical measures for both lateral and medial condyles at most applied strains and directions of applied shear. These results suggest that large-strain analyses account for nonlinear, anisotropic and location-dependent effects not fully realized at small strains. Our findings may inspire new experiments and models that consider anisotropy and heterogeneity of cartilage in ways previously ignored.

Chondrocytes depend upon solute transport within the avascular extracellular matrix of articular cartilage for many of their biological activities. Alterations to solute transport parameters may therefore mediate the cell response to tissue compression. While interstitial solute transport may be supplemented by convection during dynamic tissue compression, matrix compression is also associated with decreased diffusivities. Such trade-offs between increased convection and decreased diffusivities of solutes in dynamically compressed cartilage remain largely unexplored. We measured diffusion and convection coefficients of a wide range of solutes in mature bovine cartilage explant disks subjected to radially unconfined axial ramp compression and release. Solutes included approximately 500 Da fluorophores bearing positive and negative charges, and 10 kDa dextrans bearing positive, neutral, and negative charges. Significantly positive values of convection coefficients were measured for several different solutes. Findings therefore support a role for solute convection in mediating the cartilage biological response to dynamic compression.

The biomechanical response of articular cartilage to a wide range of impact loading rates was investigated for stress magnitudes that exist during joint trauma. Viable, intact bovine cartilage explants were impacted in confined compression with stress rates of 25, 50, 130 and 1000 MPa/s and stress magnitudes of 10, 20, 30 and 40 MPa. Water loss, cell viability, dynamic impact modulus (DIM) and matrix deformation were measured. Under all loading conditions the water loss was small (approximately 15%); water loss increased linearly with increasing peak stress and decreased exponentially with increasing stress rate. Cell death was localized within the superficial zone (or =12% of total tissue thickness); the depth of cell death from the articular surface increased with peak stress and decreased with increasing stress rate. The DIM increased (200-700 MPa) and matrix deformation decreased with increasing stress rate. Initial water and proteoglycan (PG) content had a weak, yet significant influence on water loss, cell death and DIM. However, the significance of the inhomogeneous structure and composition of the cartilage matrix was accentuated when explants impacted on the deep zone had less water loss and matrix deformation, higher DIM, and no cell death compared to explants impacted on the articular surface. The mechano-biological response of articular cartilage depended on magnitude and rate of impact loading.

Under physiological conditions of loading, articular cartilage is subjected to both compressive strains, normal to the articular surface, and tensile strains, tangential to the articular surface. Previous studies have shown that articular cartilage exhibits a much higher modulus in tension than in compression, and theoretical analyses have suggested that this tension-compression nonlinearity enhances the magnitude of interstitial fluid pressurization during loading in unconfined compression, above a theoretical threshold of 33% of the average applied stress. The first hypothesis of this experimental study is that the peak fluid load support in unconfined compression is significantly greater than the 33% theoretical limit predicted for porous permeable tissues modeled with equal moduli in tension and compression. The second hypothesis is that the peak fluid load support is higher at the articular surface side of the tissue samples than near the deep zone, because the disparity between the tensile and compressive moduli is greater at the surface zone. Ten human cartilage samples from six patellofemoral joints, and 10 bovine cartilage specimens from three calf patellofemoral joints were tested in unconfined compression. The peak fluid load support was measured at 79 +/- 11% and 69 +/- 15% at the articular surface and deep zone of human cartilage, respectively, and at 94 +/- 4% and 71 +/- 8% at the articular surface and deep zone of bovine calf cartilage, respectively. Statistical analyses confirmed both hypotheses of this study. These experimental results suggest that the tension-compression nonlinearity of cartilage is an essential functional property of the tissue which makes interstitial fluid pressurization the dominant mechanism of load support in articular cartilage.

The precise nature of the material symmetry of articular cartilage in compression remains to be elucidated. The primary objective of this study was to determine the equilibrium compressive Young's moduli and Poisson's ratios of bovine cartilage along multiple directions (parallel and perpendicular to the split line direction, and normal to the articular surface) by loading small cubic specimens (0.9×0.9×0.8 mm, n=15) in unconfined compression, with the expectation that the material symmetry of cartilage could be determined more accurately with the help of a more complete set of material properties. The second objective was to investigate how the tension-compression nonlinearity of cartilage might alter the interpretation of material symmetry. Optimized digital image correlation was used to accurately determine the resultant strain fields within the specimens under loading. Experimental results demonstrated that neither the Young's moduli nor the Poisson's ratios exhibit the same values when measured along the three loading directions. The main findings of this study are that the framework of linear orthotropic elasticity (as well as higher symmetries of linear elasticity) is not suitable to describe the equilibrium response of articular cartilage nor characterize its material symmetry; a framework which accounts for the distinctly different responses of cartilage in tension and compression is more suitable for describing the equilibrium response of cartilage; within this framework, cartilage exhibits no lower than orthotropic symmetry.

Interstitial fluid pressurization has long been hypothesized to play a fundamental role in the load support mechanism and frictional response of articular cartilage. However, to date, few experimental studies have been performed to verify this hypothesis from direct measurements. The first objective of this study was to investigate experimentally the hypothesis that cartilage interstitial fluid pressurization does support the great majority of the applied load, in the testing configurations of confined compression creep and stress relaxation. The second objective was to investigate the hypothesis that the experimentally observed interstitial fluid pressurization could also be predicted using the linear biphasic theory of Mow et al. (J. Biomech. Engng ASME, 102, 73-84, 1980). Fourteen bovine cartilage samples were tested in a confined compression chamber fitted with a microchip piezoresistive transducer to measure interstitial fluid pressure, while simultaneously measuring (during stress relaxation) or prescribing (during creep) the total stress. It was found that interstitial fluid pressure supported more than 90% of the total stress for durations as long as 725 +/- 248 s during stress relaxation (mean +/- S.D., n = 7), and 404 +/- 229 s during creep (n = 7). When comparing experimental measurements of the time-varying interstitial fluid pressure against predictions from the linear biphasic theory, nonlinear coefficients of determination r2 = 0.871 +/- 0.086 (stress relaxation) and r2 = 0.941 +/- 0.061 (creep) were found. The results of this study provide some of the most direct evidence to date that interstitial fluid pressurization plays a fundamental role in cartilage mechanics; they also indicate that the mechanism of fluid load support in cartilage can be properly predicted from theory.

Cartilage lesions change the microenvironment of cells and may accelerate cartilage degradation through catabolic responses from chondrocytes. In this study, we investigated the effects of structural integrity of the extracellular matrix (ECM) on chondrocytes by comparing the mechanics of cells surrounded by an intact ECM with cells close to a cartilage lesion using experimental and numerical methods. Experimentally, 15% nominal compression was applied to bovine cartilage tissues using a light-transmissible compression system. Target cells in the intact ECM and near lesions were imaged by dual-photon microscopy. Changes in cell morphology (N(cell)=32 for both ECM conditions) were quantified. A two-scale (tissue level and cell level) Finite Element (FE) model was also developed. A 15% nominal compression was applied to a non-linear, biphasic tissue model with the corresponding cell level models studied at different radial locations from the centre of the sample in the transient phase and at steady state. We studied the Green-Lagrange strains in the tissue and cells. Experimental and theoretical results indicated that cells near lesions deform less axially than chondrocytes in the intact ECM at steady state. However, cells near lesions experienced large tensile strains in the principal height direction, which are likely associated with non-uniform tissue radial bulging. Previous experiments showed that tensile strains of high magnitude cause an up-regulation of digestive enzyme gene expressions. Therefore, we propose that cartilage degradation near tissue lesions may be due to the large tensile strains in the principal height direction applied to cells, thus leading to an up-regulation of catabolic factors.

This study investigated the potential use of static osmotic loading as a cartilage tissue engineering strategy for growing clinically relevant grafts from either synovium-derived stem cells (SDSCs) or chondrocytes. Bovine SDSCs and chondrocytes were individually encapsulated in 2% w/v agarose and divided into chondrogenic media of osmolarities 300 (hypotonic), 330 (isotonic), and 400 (hypertonic, physiologic) mOsM for up to 7 weeks. The application of hypertonic media to constructs comprised of SDSCs or chondrocytes led to increased mechanical properties as compared to hypotonic (300 mOsM) or isotonic (330 mOsM) media (p

This study presents direct experimental evidence for assessing the electrostatic and non-electrostatic contributions of proteoglycans to the compressive equilibrium modulus of bovine articular cartilage. Immature and mature bovine cartilage samples were tested in unconfined compression and their depth-dependent equilibrium compressive modulus was determined using strain measurements with digital image correlation analysis. The electrostatic contribution was assessed by testing samples in isotonic and hypertonic saline; the combined contribution was assessed by testing untreated and proteoglycan-depleted samples. Though it is well recognized that proteoglycans contribute significantly to the compressive stiffness of cartilage, results demonstrate that the combined electrostatic and non-electrostatic contributions may add up to more than 98% of the modulus, a magnitude not previously appreciated. Of this contribution, about two thirds arises from electrostatic effects. The compressive modulus of the proteoglycan-depleted cartilage matrix may be as low as 3kPa, representing less than 2% of the normal tissue modulus; experimental evidence also confirms that the collagen matrix in digested cartilage may buckle under compressive strains, resulting in crimping patterns. Thus, it is reasonable to model the collagen as a fibrillar matrix that can sustain only tension. This study also demonstrates that residual stresses in cartilage do not arise exclusively from proteoglycans, since cartilage remains curled relative to its in situ geometry even after proteoglycan depletion. These increased insights on the structure-function relationships of cartilage can lead to improved constitutive models and a better understanding of the response of cartilage to physiological loading conditions.