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 (Young's modulus, aggregate modulus and Poisson's 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 Poisson's ratio of cartilage were also carried out. Mean values of the Young's modulus (assessed from the unconfined compression test) were 0.80+/-0.33, 0.57+/-0.17 and 0.31+/-0.18MPa and of the Poisson's 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 Young's modulus values than the unconfined compression tests. In indentation, values of the Young's modulus were independent of the indenter diameter only in the humeral cartilage. The mean values of the Poisson's ratio, obtained indirectly using the mathematical relation between the Young's 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 Poisson's 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 Young's 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.     

Mechanical stimulation to stimulate formation of a physiological collagen architecture in tissue-engineered cartilage: a numerical study

The load-bearing capacity of today's tissue-engineered (TE) cartilage is insufficient. The arcade-like collagen network in native cartilage plays an important role in its load-bearing properties. Inducing the formation of such collagen architecture in engineered cartilage can, therefore, enhance mechanical properties of TE cartilage. Considering the well-defined relationship between tensile strains and collagen alignment in the literature, we assume that cues for inducing this orientation should come from mechanical loading. In this study, strain fields prescribed by loading conditions of unconfined compression, sliding indentation and a novel loading regime of compression-sliding indentation are numerically evaluated to assess the probability that these would trigger a physiological collagen architecture. Results suggest that sliding indentation is likely to stimulate the formation of an appropriate superficial zone with parallel fibres. Adding lateral compression may stimulate the formation of a deep zone with perpendicularly aligned fibres. These insights may be used to improve loading conditions for cartilage tissue engineering.     

The Mechanical Behaviour of Chondrocytes Predicted with a Micro-structural Model of Articular Cartilage

The integrity of articular cartilage depends on the proper functioning and mechanical stimulation of chondrocytes, the cells that synthesize extracellular matrix and maintain tissue health. The biosynthetic activity of chondrocytes is influenced by genetic factors, environmental influences, extracellular matrix composition, and mechanical factors. The mechanical environment of chondrocytes is believed to be an important determinant for joint health, and chondrocyte deformation in response to mechanical loading is speculated to be an important regulator of metabolic activity. In previous studies of chondrocyte deformation, articular cartilage was described as a biphasic material consisting of a homogeneous, isotropic, linearly elastic solid phase, and an inviscid fluid phase. However, articular cartilage is known to be anisotropic and inhomogeneous across its depth. Therefore, isotropic and homogeneous models cannot make appropriate predictions for tissue and cell stresses and strains. Here, we modelled articular cartilage as a transversely isotropic, inhomogeneous (TI) material in which the anisotropy and inhomogeneity arose naturally from the microstructure of the depth-dependent collagen fibril orientation and volumetric fraction, as well as the chondrocyte shape and volumetric fraction. The purpose of this study was to analyse the deformation behaviour of chondrocytes using the TI model of articular cartilage. In order to evaluate our model against experimental results, we simulated indentation and unconfined compression tests for nominal compressions of 15%. Chondrocyte deformations were analysed as a function of location within the tissue. The TI model predicted a non-uniform behaviour across tissue depth: in indentation testing, cell height decreased by 43% in the superficial zone and between 11 and 29% in the deep zone. In unconfined compression testing, cell height decreased by 32% in the superficial zone, 25% in the middle, and 18% in the deep zones. This predicted non-uniformity is in agreement with experimental studies. The novelty of this study is the use of a cartilage material model accounting for the intrinsic inhomogeneity and anisotropy of cartilage caused by its microstructure.     

Stress-relaxation of human patellar articular cartilage in unconfined compression: prediction of mechanical response by tissue composition and structure

Mechanical properties of articular cartilage are controlled by tissue composition and structure. Cartilage function is sensitively altered during tissue degeneration, in osteoarthritis (OA). However, mechanical properties of the tissue cannot be determined non-invasively. In the present study, we evaluate the feasibility to predict, without mechanical testing, the stress-relaxation response of human articular cartilage under unconfined compression. This is carried out by combining microscopic and biochemical analyses with composition-based mathematical modeling. Cartilage samples from five cadaver patellae were mechanically tested under unconfined compression. Depth-dependent collagen content and fibril orientation, as well as proteoglycan and water content were derived by combining Fourier transform infrared imaging, biochemical analyses and polarized light microscopy. Finite element models were constructed for each sample in unconfined compression geometry. First, composition-based fibril-reinforced poroviscoelastic swelling models, including composition and structure obtained from microscopical and biochemical analyses were fitted to experimental stress-relaxation responses of three samples. Subsequently, optimized values of model constants, as well as compositional and structural parameters were implemented in the models of two additional samples to validate the optimization. Theoretical stress-relaxation curves agreed with the experimental tests (R=0.95-0.99). Using the optimized values of mechanical parameters, as well as composition and structure of additional samples, we were able to predict their mechanical behavior in unconfined compression, without mechanical testing (R=0.98). Our results suggest that specific information on tissue composition and structure might enable assessment of cartilage mechanics without mechanical testing.     

A cross-validation of the biphasic poroviscoelastic model of articular cartilage in unconfined compression, indentation, and confined compression

The biphasic poroviscoelastic (BPVE) model was curve fit to the simultaneous relaxation of reaction force and lateral displacement exhibited by articular cartilage in unconfined compression (n=18). Model predictions were also made for the relaxation observed in reaction force during indentation with a porous plane-ended metal indenter (n=4), indentation with a nonporous plane ended metal indenter (n=4), and during confined compression (n=4). Each prediction was made using material parameters resulting from curve fits of the unconfined compression response of the same tissue. The BPVE model was able to account for both the reaction force and the lateral displacement during unconfined compression very well. Furthermore, model predictions for both indentation and confined compression also followed the experimental data well. These results provide substantial evidence for the efficacy of the biphasic poroviscoelastic model for articular cartilage, as no successful cross-validation of a model simulation has been demonstrated using other mathematical models.     

The role of viscoelasticity of collagen fibers in articular cartilage: theory and numerical formulation

The relative importance of fluid-dependent and fluid-independent transient mechanical behavior in articular cartilage was examined for tensile and unconfined compression testing using a fibril reinforced model. The collagen matrix of articular cartilage was modeled as viscoelastic using a quasi-linear viscoelastic formulation with strain-dependent elastic modulus, while the proteoglycan matrix was considered as linearly elastic. The collagen viscoelastic properties were obtained by fitting experimental data from a tensile test. These properties were used to investigate unconfined compression testing, and the sensitivity of the properties was also explored. It was predicted that the stress relaxation observed in tensile tests was not caused by fluid pressurization at the macroscopic level. A multi-step tensile stress relaxation test could be approximated using a hereditary integral in which the elastic fibrillar modulus was taken to be a linear function of the fibrillar strain. Applying the same formulation to the radial fibers in unconfined compression, stress relaxation could not be simulated if fluid pressurization were absent. Collagen viscoelasticity was found to slightly weaken fluid pressurization in unconfined compression, and this effect was relatively more significant at moderate strain rates. Therefore, collagen viscoelasticity appears to play an import role in articular cartilage in tensile testing, while fluid pressurization dominates the transient mechanical behavior in compression. Collagen viscoelasticity plays a minor role in the mechanical response of cartilage in unconfined compression if significant fluid flow is present.     

Biomechanical properties of human articular cartilage under compressive loads

The function of articular cartilage is to support and distribute loads and to provide lubrication in the diarthrodial joints. Cartilage function is described by proper mechanical and rheological properties, strain and depth-dependent, which are not completely assessed. Unconfined and confined compression are commonly used to evaluate the Young's modulus (E) and the aggregate modulus (H(A)), respectively. The Poisson's ratio (nu) can be calculated indirectly from the equilibrium compression data, or using the biphasic indentation technique; it has recently been optically evaluated by using video microscopy during unconfined compression. The transient response of articular cartilage during confined compression depends on its permeability k; a constant value of k can be easily identified by a simple analytical model of confined compression tests, whereas more complex models or direct measurements (permeation tests) are needed to study the permeability dependence on deformation. A poroelastic finite element model of articular cartilage was developed for this purpose. The elastic parameters (E,nu) of the model were evaluated performing unconfined compression creep tests on human articular cartilage disks, whereas k was identified from the confined test response. Our combined experimental and computational method can be used to identify the parameters that define the permeability dependence on deformation, as a function of depth from articular surface.     

The generalized triphasic correspondence principle for simultaneous determination of the mechanical properties and proteoglycan content of articular cartilage by indentation

The triphasic mixture theory has been used to describe the mechanical and physicochemical behaviors of articular cartilage under some specialized loading conditions. However, the mathematical complexities of this theory have limited its applications for theoretical analyses of experimental studies and models for predicting cartilage and other biological tissues' deformational behaviors. A generalized correspondence principle has been established in the present study, and this principle shows that the equilibrium deformational behavior of a charged-hydrated material under loading is identical to that of an elastic medium without charge. A set of explicit formulas has been derived to correlate the mechanical properties of an equivalent material with the intrinsic elastic moduli, fixed charge density and free-ion concentration within the cartilage tissue. The validity of these formulas is independent of the deformation state of the elastic solid matrix under an infinitesimal strain. Therefore they can be employed for any loading conditions, such as confined or unconfined compression, tension, and indentation tests, etc. In the current study, the fixed charge density of bovine cartilage is determined from the indentation creep data using this generalized correspondence principle. The proteoglycan content results were then compared with those from biochemical assay, yielding a linear regression slope of 1.034. Additionally a correspondence principle within a framework of cubic symmetry and a bilinear response in tension-compression (the conewise linear elasticity model) has also been developed to demonstrate the potential application of current methodology for inhomogeneous, anisotropic and nonlinear situations.     

A linearized formulation of triphasic mixture theory for articular cartilage,and its application to indentation analysis

The negative charges on proteoglycans significantly affect the mechanical behaviors of articular cartilage. Mixture theories, such as the triphasic theory, can describe quantitatively how this charged nature contributes to the mechano-electrochemical behaviors of such tissue. However, the mathematical complexity of the theory has hindered its application to complicated loading profiles, e.g., indentation or other multi-dimensional configurations. In this study, the governing equations of triphasic mixture theory for soft tissue were linearized and dramatically simplified by using a regular perturbation method and the use of two potential functions. We showed that this new formulation can be used for any axisymmetric problem, such as confined or unconfined compressions, hydraulic perfusion, and indentation. A finite difference numerical program was further developed to calculate the deformational, electrical, and flow behaviors inside the articular cartilage under indentation. The calculated tissue response was highly consistent with the data from indentation experiments (our own and those reported in the literature). It was found that the charged nature of proteoglycans can increase the apparent stiffness of the solid matrix and lessen the viscous effect introduced by fluid flow. The effects of geometric and physical properties of indenter tip, cartilage thickness, and that of the electro-chemical properties of cartilage on the resulting deformation and fluid pressure fields across the tissue were also investigated and presented. These results have implications for studying chondrocyte mechanotransduction in different cartilage zones and for tissue engineering designs or in vivo cartilage repair.     

Experimental verification of the roles of intrinsic matrix viscoelasticity and tension-compression nonlinearity in the biphasic response of cartilage

A biphasic-CLE-QLV model proposed in our recent study [2001, J. Biomech. Eng., 123, pp. 410-417] extended the biphasic theory of Mow et al. [1980, J. Biomech. Eng., 102, pp. 73-84] to include both tension-compression nonlinearity and intrinsic viscoelasticity of the cartilage solid matrix by incorporating it with the conewise linear elasticity (CLE) model [1995, J. Elasticity, 37, pp. 1-38] and the quasi-linear viscoelasticity (QLV) model [Biomechanics: Its foundations and objectives, Prentice Hall, Englewood Cliffs, 1972]. This model demonstrates that a simultaneous prediction of compression and tension experiments of articular cartilage, under stress-relaxation and dynamic loading, can be achieved when properly taking into account both flow-dependent and flow-independent viscoelastic effects, as well as tension-compression nonlinearity. The objective of this study is to directly test this biphasic-CLE-QLV model against experimental data from unconfined compression stress-relaxation tests at slow and fast strain rates as well as dynamic loading. Twelve full-thickness cartilage cylindrical plugs were harvested from six bovine glenohumeral joints and multiple confined and unconfined compression stress-relaxation tests were performed on each specimen. The material properties of specimens were determined by curve-fitting the experimental results from the confined and unconfined compression stress relaxation tests. The findings of this study demonstrate that the biphasic-CLE-QLV model is able to describe the strain-rate-dependent mechanical behaviors of articular cartilage in unconfined compression as attested by good agreements between experimental and theoretical curvefits (r2 = 0.966 +/- 0.032 for testing at slow strain rate; r2 = 0.998 +/- 0.002 for testing at fast strain rate) and predictions of the dynamic response (r2 = 0.91 +/- 0.06). This experimental study also provides supporting evidence for the hypothesis that both tension-compression nonlinearity and intrinsic viscoelasticity of the solid matrix of cartilage are necessary for modeling the transient and equilibrium responses of this tissue in tension and compression. Furthermore, the biphasic-CLE-QLV model can produce better predictions of the dynamic modulus of cartilage in unconfined dynamic compression than the biphasic-CLE and biphasic poroviscoelastic models, indicating that intrinsic viscoelasticity and tension-compression nonlinearity of articular cartilage may play important roles in the load-support mechanism of cartilage under physiologic loading.     

Mechano-acoustic determination of Young's modulus of articular cartilage

The compressive stiffness of an elastic material is traditionally characterized by its Young's modulus. Young's modulus of articular cartilage can be directly measured using unconfined compression geometry by assuming the cartilage to be homogeneous and isotropic. In isotropic materials, Young's modulus can also be determined acoustically by the measurement of sound speed and density of the material. In the present study, acoustic and mechanical techniques, feasible for in vivo measurements, were investigated to quantify the static and dynamic compressive stiffness of bovine articular cartilage in situ. Ultrasound reflection from the cartilage surface, as well as the dynamic modulus were determined with the recently developed ultrasound indentation instrument and compared with the reference mechanical and ultrasound speed measurements in unconfined compression (n=72). In addition, the applicability of manual creep measurements with the ultrasound indentation instrument was evaluated both experimentally and numerically. Our experimental results indicated that the sound speed could predict 47% and 53% of the variation in the Young's modulus and dynamic modulus of cartilage, respectively. The dynamic modulus, as determined manually with the ultrasound indentation instrument, showed significant linear correlations with the reference Young's modulus (r(2)=0.445, p<0.01, n=70) and dynamic modulus (r(2)=0.779, p<0.01, n=70) of the cartilage. Numerical analyses indicated that the creep measurements, conducted manually with the ultrasound indentation instrument, were sensitive to changes in Young's modulus and permeability of the tissue, and were significantly influenced by the tissue thickness. We conclude that acoustic parameters, i.e. ultrasound speed and reflection, are indicative to the intrinsic mechanical properties of the articular cartilage. Ultrasound indentation instrument, when further developed, provides an applicable tool for the in vivo detection of cartilage mechano-acoustic properties. These techniques could promote the diagnostics of osteoarthrosis.     

Biomechanical properties of knee articular cartilage

Structure and properties of knee articular cartilage are adapted to stresses exposed on it during physiological activities. In this study, we describe site- and depth-dependence of the biomechanical properties of bovine knee articular cartilage. We also investigate the effects of tissue structure and composition on the biomechanical parameters as well as characterize experimentally and numerically the compression-tension nonlinearity of the cartilage matrix. In vitro mechano-optical measurements of articular cartilage in unconfined compression geometry are conducted to obtain material parameters, such as thickness, Young's and aggregate modulus or Poisson's ratio of the tissue. The experimental results revealed significant site- and depth-dependent variations in recorded parameters. After enzymatic modification of matrix collagen or proteoglycans our results show that collagen primarily controls the dynamic tissue response while proteoglycans affect more the static properties. Experimental measurements in compression and tension suggest a nonlinear compression-tension behavior of articular cartilage in the direction perpendicular to articular surface. Fibril reinforced poroelastic finite element model was used to capture the experimentally found compression-tension nonlinearity of articular cartilage.     

Tensile and compressive properties of healthy and osteoarthritic human articular cartilage

Osteoarthritis (OA) is a disease affecting articular cartilage and the underlying bone, resulting from many biological and mechanical interacting factors which change the extracellular matrix (ECM) and cells and lead to increasing levels of cartilage degeneration, like softening, fibrillation, ulceration and cartilage loss. The early diagnosis of the disease is fundamental to prevent pain, further tissue degeneration and reduce hospital costs. Although morphological modifications can be detected by modern non-invasive diagnostic techniques, they may not be evident in the early stages of OA. The mechanical properties of articular cartilage are related to its composition and structure and are sensitive to even small changes in the ECM that could occur in early OA. The aim of the present study was to compare the mechanical properties of healthy and OA cartilage using a combined experimental-numerical approach. Experimental assessments consisted of step wise confined and unconfined compression and tension stress relaxation tests on disks (for compression) or strips (for tension) of cartilage obtained from human femoral heads discarded from the operating room after total hip replacement. The numerical model was based on the biphasic theory and included the tension-compression non-linearity. Considering OA samples vs normal samples, the static compressive modulus was 55-68% lower, the permeability was 60-80% higher, the dynamic compressive modulus was 59-64% lower, the static tension modulus was 72-83% lower. The model successfully simulated the experimental tests performed on healthy and OA cartilage and was used in combination with the experimental tests to evaluate the role of different ECM components in the mechanical response of normal and OA cartilage.     

Strain-rate dependent stiffness of articular cartilage in unconfined compression

The stiffness of articular cartilage is a nonlinear function of the strain amplitude and strain rate as well as the loading history, as a consequence of the flow of interstitial water and the stiffening of the collagen fibril network. This paper presents a full investigation of the interplay between the fluid kinetics and fibril stiffening of unconfined cartilage disks by analyzing over 200 cases with diverse material properties. The lower and upper elastic limits of the stress (under a given strain) are uniquely established by the instantaneous and equilibrium stiffness (obtained numerically for finite deformations and analytically for small deformations). These limits could be used to determine safe loading protocols in order that the stress in each solid constituent remains within its own elastic limit. For a given compressive strain applied at a low rate, the loading is close to the lower limit and is mostly borne directly by the solid constituents (with little contribution from the fluid). In contrast, however in case of faster compression, the extra loading is predominantly transported to the fibrillar matrix via rising fluid pressure with little increase of stress in the nonfibrillar matrix. The fibrillar matrix absorbs the loading increment by self-stiffening: the quicker the loading the faster the fibril stiffening until the upper elastic loading limit is reached. This self-protective mechanism prevents cartilage from damage since the fibrils are strong in tension. The present work demonstrates the ability of the fibril reinfored poroelastic models to describe the strain rate dependent behavior of articular cartilage in unconfined compression using a mechanism of fibril stiffening mainly induced by the fluid flow.     

Importance of collagen orientation and depth-dependent fixed charge densities of cartilage on mechanical behavior of chondrocytes

The collagen network and proteoglycan matrix of articular cartilage are thought to play an important role in controlling the stresses and strains in and around chondrocytes, in regulating the biosynthesis of the solid matrix, and consequently in maintaining the health of diarthrodial joints. Understanding the detailed effects of the mechanical environment of chondrocytes on cell behavior is therefore essential for the study of the development, adaptation, and degeneration of articular cartilage. Recent progress in macroscopic models has improved our understanding of depth-dependent properties of cartilage. However, none of the previous works considered the effect of realistic collagen orientation or depth-dependent negative charges in microscopic models of chondrocyte mechanics. The aim of this study was to investigate the effects of the collagen network and fixed charge densities of cartilage on the mechanical environment of the chondrocytes in a depth-dependent manner. We developed an anisotropic, inhomogeneous, microstructural fibril-reinforced finite element model of articular cartilage for application in unconfined compression. The model consisted of the extracellular matrix and chondrocytes located in the superficial, middle, and deep zones. Chondrocytes were surrounded by a pericellular matrix and were assumed spherical prior to tissue swelling and load application. Material properties of the chondrocytes, pericellular matrix, and extracellular matrix were obtained from the literature. The loading protocol included a free swelling step followed by a stress-relaxation step. Results from traditional isotropic and transversely isotropic biphasic models were used for comparison with predictions from the current model. In the superficial zone, cell shapes changed from rounded to elliptic after free swelling. The stresses and strains as well as fluid flow in cells were greatly affected by the modulus of the collagen network. The fixed charge density of the chondrocytes, pericellular matrix, and extracellular matrix primarily affected the aspect ratios (height/width) and the solid matrix stresses of cells. The mechanical responses of the cells were strongly location and time dependent. The current model highlights that the collagen orientation and the depth-dependent negative fixed charge densities of articular cartilage have a great effect in modulating the mechanical environment in the vicinity of chondrocytes, and it provides an important improvement over earlier models in describing the possible pathways from loading of articular cartilage to the mechanical and biological responses of chondrocytes.     

Modelling of location- and time-dependent deformation of chondrocytes during cartilage loading.

Experimental evidence suggests that the biosynthetic activity of chondrocytes is regulated primarily by the mechanical environment. In order to study the mechanisms underlying remodeling, adaptation, and degeneration of articular cartilage in a joint subjected to changing loads, it is important to know the time-dependent fluid pressure and stress-strain state in chondrocytes. The purpose of the present study was to develop a theoretical model to simulate the mechanical behaviour of articular cartilage and to describe the time-dependent stress-strain state and fluid pressure distribution in chondrocytes during cartilage deformation. It was assumed that the volume occupied by the chondrocytes is small and that cartilage can be treated as a macroscopically homogenized material with effective material properties which depend on the material properties of the cells and matrix and the volumetric fraction of the cells. Model predictions on the time-dependent distribution of fluid pressure and stress and on the time-dependent cell deformation during confined and unconfined compression tests agree with previous theoretical predictions and experimental observations. The proposed model supplies the tools to study the mechanisms of degeneration, adaptation and remodelling of cartilage associated with cell loading and deformation.     

Rapid regulation of collagen but not metalloproteinase 1, 3, 13, 14 and tissue inhibitor of metalloproteinase 1, 2, 3 expression in response to mechanical loading of cartilage explants in vitro

This study analyzes the molecular response of articular chondrocytes to short-term mechanical loading with a special focus on gene expression of molecules relevant for matrix turnover. Porcine cartilage explants were exposed to static and dynamic unconfined compression and viability of chondrocytes was assessed to define physiologic loading conditions. Cell death in the superficial layer correlated with mechanical loading and occurred at peak stresses >or=6 MPa and a cartilage compression above 45%. Chondrocytes in native cartilage matrix responded to dynamic loading by rapid and highly specific suppression of collagen expression. mRNA levels dropped 11-fold (collagen 2; 6 MPa, P=0.009) or 14-fold (collagen 1; 3 and 6 MPa, P=0.009) while levels of aggrecan, tenascin-c, matrix metalloproteinases (MMP1, 3, 13, 14), and their inhibitors (TIMP1-3) did not change significantly. Thus, dynamic mechanical loading rapidly shifted the balance between collagen and aggrecan/tenascin/MMP/TIMP expression. A better knowledge of the chondrocyte response to mechanical stress may improve our understanding of mechanically induced osteoarthrits.     

Tissue engineering of cartilage using a mechanobioreactor exerting simultaneous mechanical shear and compression to simulate the rolling action of articular joints

The effect of dynamic mechanical shear and compression on the synthesis of human tissue‐engineered cartilage was investigated using a mechanobioreactor capable of simulating the rolling action of articular joints in a mixed fluid environment. Human chondrocytes seeded into polyglycolic acid (PGA) mesh or PGA–alginate scaffolds were precultured in shaking T‐flasks or recirculation perfusion bioreactors for 2.5 or 4 weeks prior to mechanical stimulation in the mechanobioreactor. Constructs were subjected to intermittent unconfined shear and compressive loading at a frequency of 0.05 Hz using a peak‐to‐peak compressive strain amplitude of 2.2% superimposed on a static axial compressive strain of 6.5%. The mechanical treatment was carried out for up to 2.5 weeks using a loading regime of 10 min duration each day with the direction of the shear forces reversed after 5 min and release of all loading at the end of the daily treatment period. Compared with shaking T‐flasks and mechanobioreactor control cultures without loading, mechanical treatment improved the amount and quality of cartilage produced. On a per cell basis, synthesis of both major structural components of cartilage, glycosaminoglycan (GAG) and collagen type II, was enhanced substantially by up to 5.3‐ and 10‐fold, respectively, depending on the scaffold type and seeding cell density. Levels of collagen type II as a percentage of total collagen were also increased after mechanical treatment by up to 3.4‐fold in PGA constructs. Mechanical treatment had a less pronounced effect on the composition of constructs precultured in perfusion bioreactors compared with perfusion culture controls. This work demonstrates that the quality of tissue‐engineered cartilage can be enhanced significantly by application of simultaneous dynamic mechanical shear and compression, with the greatest benefits evident for synthesis of collagen type II. Biotechnol. Bioeng. 2012; 109:1060–1073. © 2011 Wiley Periodicals, Inc.