In Vivo Dynamic Deformation of Articular Cartilage in Intact Joints Loaded by Controlled Muscular Contractions

When synovial joints are loaded, the articular cartilage and the cells residing in it deform. Cartilage deformation has been related to structural tissue damage, and cell deformation has been associated with cell signalling and corresponding anabolic and catabolic responses. Despite the acknowledged importance of cartilage and cell deformation, there are no dynamic data on these measures from joints of live animals using muscular load application. Research in this area has typically been done using confined and unconfined loading configurations and indentation testing. These loading conditions can be well controlled and allow for accurate measurements of cartilage and cell deformations, but they have little to do with the contact mechanics occurring in a joint where non-congruent cartilage surfaces with different material and functional properties are pressed against each other by muscular forces. The aim of this study was to measure in vivo, real time articular cartilage deformations for precisely controlled static and dynamic muscular loading conditions in the knees of mice. Fifty and 80% of the maximal knee extensor muscular force (equivalent to approximately 0.4N and 0.6N) produced average peak articular cartilage strains of 10.5±1.0% and 18.3±1.3% (Mean ± SD), respectively, during 8s contractions. A sequence of 15 repeat, isometric muscular contractions (0.5s on, 3.5s off) of 50% and 80% of maximal muscular force produced cartilage strains of 3.0±1.1% and 9.6±1.5% (Mean ± SD) on the femoral condyles of the mouse knee. Cartilage thickness recovery following mechanical compression was highly viscoelastic and took almost 50s following force removal in the static tests.     

Influence of stress rate on water loss, matrix deformation and chondrocyte viability in impacted articular cartilage

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.     

The effect of loading rate on the development of early damage in articular cartilage

Experimental reports suggest that cartilage damage depends on strain magnitude. Additionally, because of its poro-viscoelastic nature, strain magnitude in cartilage can depend on strain rate. The present study explores whether cartilage damage may develop dependent on strain rate, even when the presented damage numerical model is strain-dependent but not strain-rate-dependent. So far no experiments have been distinguished whether rate-dependent cartilage damage occurs in the collagen or in the non-fibrillar network. Thus, this research presents a finite element analysis model where, among others, collagen and non-fibrillar matrix are incorporated as well as a strain-dependent damage mechanism for these components. Collagen and non-fibrillar matrix stiffness decrease when a given strain is reached until complete failure upon reaching a maximum strain. With such model, indentation experiments at increasing strain rates were simulated on cartilage plugs and damage development was monitored over time. Collagen damage increased with increasing strain rate from 21 to 42 %. In contrast, damage in the non-fibrillar matrix decreased with increasing strain rates from 72 to 34 %. Damage started to develop at a depth of approximately 20 % of the sample height, and this was more pronounced for the slow and modest loading rates. However, the most severe damage at the end of the compression step occurred at the surface for the plugs subjected to 120 mm/min strain rate. In conclusion, the present study confirms that the location and magnitude of damage in cartilage may be strongly dependent on strain rate, even when damage occurs solely through a strain-dependent damage mechanism.     

Optical determination of anisotropic material properties of bovine articular cartilage in compression

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 x 0.9 x 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.     

Contribution of tissue composition and structure to mechanical response of articular cartilage under different loading geometries and strain rates

Mechanical function of articular cartilage in joints between articulating bones is dependent on the composition and structure of the tissue. The mechanical properties of articular cartilage are traditionally tested in compression using one of the three loading geometries, i.e., confined compression, unconfined compression or indentation. The aim of this study was to utilize a composition-based finite element model in combination with a fractional factorial design to determine the importance of different cartilage constituents in the mechanical response of the tissue, and to compare the importance of the tissue constituents with different loading geometries and loading rates. The evaluated parameters included water and collagen fraction as well as fixed charge density on cartilage surface and their slope over the tissue thickness. The thicknesses of superficial and middle zones, as based on the collagen orientation, were also included in the evaluated parameters. A three-level resolution V fractional factorial design was used. The model results showed that inhomogeneous composition plays only a minor role in indentation, though that role becomes more significant in confined compression and unconfined compression. In contrast, the collagen architecture and content had a more profound role in indentation than with two other loading geometries. These differences in the mechanical role of composition and structure between the loading geometries were emphasized at higher loading rates. These findings highlight how the results from mechanical tests of articular cartilage under different loading conditions are dependent upon tissue composition and structure.     

A finite deformation theory for cartilage and other soft hydrated connective tissues--I. Equilibrium results

The determination of valid stress-strain relations for articular cartilage under finite deformation conditions is a prerequisite for constructing models for synovial joint lubrication. Under physiological conditions of high strain rates and/or high stresses in the joint, large strains occur in cartilage. A finite deformation theory valid for describing cartilage, as well as other soft hydrated connective tissues under large loads, has been developed. This theory is based on the choice of a specific Helmholtz energy function which satisfies the generalized Coleman-Noll (GCN0) condition and the Baker-Ericksen (B-E) inequalities established in finite elasticity theory. In addition, the finite deformation biphasic theory includes the effects of strain-dependent porosity and permeability. These nonlinear effects are essential for properly describing the biomechanical behavior of articular cartilage, even when strain rates are low and strains are infinitesimal. The finite deformation theory describes the large strain behavior of cartilage observed in one-dimensional confined compression experiments at equilibrium, and it reduces to the linear biphasic theory under infinitesimal strain and slow strain rate conditions. Using this theory, we have determined the material coefficients of both human and bovine articular cartilages under large strain conditions at equilibrium. The theory compares very well with experimental results.     

Biphasic poroviscoelastic simulation of the unconfined compression of articular cartilage: II--Effect of variable strain rates

This study investigated the abilities of the linear biphasic poroviscoelastic (BPVE) model and the linear biphasic poroelastic (BPE) model to simulate the effect of variable ramp strain rates on the unconfined compression stress relaxation response of articular cartilage. Curve fitting of experimental data showed that the BPVE model was able to successfully account for the ramp strain rate-dependent viscoelastic behavior of articular cartilage under unconfined compression, while the BPE model was able to account for the complete viscoelastic response at a slow strain rate, but only the long-term viscoelastic response at faster strain rates. We concluded that the short-term viscoelastic behavior of articular cartilage, when subjected to a fast ramp strain rate, is primarily governed by a fluid flow-independent (intrinsic) viscoelastic mechanism, whereas the long-term viscoelastic behavior is governed by a fluid flow-dependent (biphasic) viscoelastic mechanism. Furthermore, a linear viscoelastic representation of the solid stress was found to be a valid model assumption for the simulation of ramp strain rate-dependent relaxation behaviors of articular cartilage within the range of ramp strain rates investigated.     

Effect of superficial collagen patterns and fibrillation of femoral articular cartilage on knee joint mechanics-a 3D finite element analysis

Collagen fibrils of articular cartilage have specific depth-dependent orientations and the fibrils bend in the cartilage surface to exhibit split-lines. Fibrillation of superficial collagen takes place in osteoarthritis. We aimed to investigate the effect of superficial collagen fibril patterns and collagen fibrillation of cartilage on stresses and strains within a knee joint. A 3D finite element model of a knee joint with cartilage and menisci was constructed based on magnetic resonance imaging. The fibril-reinforced poroviscoelastic material properties with depth-dependent collagen orientations and split-line patterns were included in the model. The effects of joint loading on stresses and strains in cartilage with various split-line patterns and medial collagen fibrillation were simulated under axial impact loading of 1000 N. In the model, the collagen fibrils resisted strains along the split-line directions. This increased also stresses along the split-lines. On the contrary, contact and pore pressures were not affected by split-line patterns. Simulated medial osteoarthritis increased tissue strains in both medial and lateral femoral condyles, and contact and pore pressures in the lateral femoral condyle. This study highlights the importance of the collagen fibril organization, especially that indicated by split-line patterns, for the weight-bearing properties of articular cartilage. Osteoarthritic changes of cartilage in the medial femoral condyle created a possible failure point in the lateral femoral condyle. This study provides further evidence on the importance of the collagen fibril organization for the optimal function of articular cartilage.     

Development of a preclinical natural porcine knee simulation model for the tribological assessment of osteochondral grafts in vitro

In order to pre-clinically evaluate the performance and efficacy of novel osteochondral interventions, physiological and clinically relevant whole joint simulation models, capable of reproducing the complex loading and motions experienced in the natural knee environment are required. The aim of this study was to develop a method for the assessment of tribological performance of osteochondral grafts within an in vitro whole natural joint simulation model.The study assessed the effects of osteochondral allograft implantation (existing surgical intervention for the repair of osteochondral defects) on the wear, deformation and damage of the opposing articular surfaces. Tribological performance of osteochondral grafts was compared to the natural joint (negative control), an injury model (focal cartilage defects) and stainless steel pins (positive controls). A recently developed method using an optical profiler (Alicona Infinite Focus G5, Alicona Imaging GmbH, Austria) was used to quantify and characterise the wear, deformation and damage occurring on the opposing articular surfaces. Allografts inserted flush with the cartilage surface had the lowest levels of wear, deformation and damage following the 2 h test; increased levels of wear, deformation and damage were observed when allografts and stainless steel pins were inserted proud of the articular surface. The method developed will be applied in future studies to assess the tribological performance of novel early stage osteochondral interventions prior to in vivo studies, investigate variation in surgical precision and aid in the development of stratified interventions for the patient population.     

The "instantaneous" deformation of cartilage: effects of collagen fiber orientation and osmotic stress

The present study was undertaken with two objectives in view. The first was to distinguish between the "instantaneous" deformation and creep of articular cartilage when subjected to a step loading in unconfined compression. This was done by observing changes in the specimen's diameter rather than its thickness. The second objective was to investigate experimentally the anisotropic behaviour of cartilage in a compressive loading mode, corresponding to the physiological situation. An apparatus was thus developed and constructed which enabled us to follow the "instantaneous" changes of the surface area of the sample as the latter was being loaded in unconfined compression. Specimens of human articular cartilage from normal femoral heads and condyles were tested. Full thickness specimens were tested with and without the underlying bone, as well as partial thickness specimens, characterizing the different zones of cartilage. Solutions of different ionic strength were used to vary the osmotic stress and specimens covering a considerable range of proteoglycan concentrations were selected. The effects of hydration and proteoglycan removal on the "instantaneous" deformation were also studied. The "instantaneous" deformation was found to be of a strongly anisotropic nature in all zones. The deformation was always smaller along the Indian-ink prick pattern than at 90 degrees to it, and this effect was most pronounced in the superficial zone of cartilage. The results reveal an analogy with the tensile properties of cartilage and indicate that the collagen network is mainly responsible for controlling the "instantaneous" deformation. The proteoglycans play an indirect role by modulating the stiffness of the collagen network through their osmotic pressure.     

Post-traumatic osteoarthritis: the role of stress induced chondrocyte damage

Post-traumatic osteoarthritis is the form of osteoarthritis (OA) that develops following joint injury. Although its end-stage is indistinguishable from idiopathic OA, many patients with post-traumatic OA are younger than those with idiopathic OA, and they have a well-defined precipitating insult. Clinical and experimental studies suggest that excessive acute impact energy or chronic mechanical overload cause the degeneration of the articular surface responsible for post-traumatic OA. Yet, the mechanisms by which excessive mechanical force causes OA remain unknown. For these reasons it has not been possible to develop effective methods of preventing or decreasing the risk of post-traumatic OA. We hypothesized that mechanical loading that exceeds the tolerance of the articular surface causes chondrocyte damage due to oxidative stress. Our in vitro tests of human articular cartilage samples showed that shear stress causes chondrocyte death and that anti-oxidants decrease the shear stress induced cell death. These observations suggest that specific patterns of loading are particularly damaging to articular surfaces and that improved treatments of joint injuries may include mechanical methods of minimizing shear stresses and biologic methods of minimizing oxidative damage.     

Effects of macro-cracks on the load bearing capacity of articular cartilage

Biomechanics and Modeling in Mechanobiology - Macro-cracks on the surface of articular cartilage are one of the hallmarks of early osteoarthritis and joint damage initiation. Macro-cracks...     

Biomechanics of osteochondral impact with cushioning and graft Insertion: Cartilage damage is correlated with delivered energy

Articular cartilage is susceptible to impact injury. Impact may occur during events ranging from trauma to surgical insertion of an OsteoChondral Graft (OCG) into an OsteoChondral Recipient site (OCR). To evaluate energy density as a mediator of cartilage damage, a specialized drop tower apparatus was used to impact adult bovine samples while measuring contact force, cartilage surface displacement, and OCG advancement. When a single impact was applied to an isolated (non-inserted) OCG, force and surface displacement each rose monotonically and then declined. In each of five sequential impacts of increasing magnitude, applied to insert an OCG into an OCR, force rose rapidly to an initial peak, with minimal OCG advancement, and then to a second prolonged peak, with distinctive oscillations. Energy delivered to cartilage was confirmed to be higher with larger drop height and mass, and found to be lower with an interposed cushion or OCG insertion into an OCR. For both single and multiple impacts, the total energy density delivered to the articular cartilage correlated to damage, quantified as total crack length. The corresponding fracture toughness of the articular cartilage was 12.0 mJ/mm². Thus, the biomechanics of OCG insertion exhibits distinctive features compared to OCG impact without insertion, with energy delivery to the articular cartilage being a factor highly correlated with damage.     

Fluid load support during localized indentation of cartilage with a spherical probe

Interstitial fluid pressurization, a consequence of a biphasic tissue structure, is essential to the load bearing and lubrication properties of articular cartilage. Focal tissue degradation may interfere with this protective mechanism, eventually leading to gross degeneration and osteoarthritis. Our long-term goal is to determine whether local contacts can be used as a means to probe local tissue integrity and functionality. In the present work, Hertzian rate-controlled microindentation was used as a model of the more complicated sliding system to directly determine the effects of contact radius and deformation rate on interstitial load support. During localized contact between a steel spherical probe and bovine articular cartilage, the equilibrium and non-equilibrium responses were well-fit by the Hertz model (R(2)>0.998) with a mean equilibrium contact modulus of 0.93 MPa. The effective contact modulus and fluid load fraction were independent of indentation depth, contact radius, and normal force; both increased monotonically with indentation rate. At 21 μm/s indentation rate, the cartilage was effectively stiffened by 6-fold with the fluid pressure supporting 85% of the contact force. The results motivated a simple analytical model that directly links the tribomechanical response (including fluid load support) and the Peclet number to measurable material properties and controllable experimental variables. This paper demonstrates that tribological contacts can be used to probe local functional properties. Such measurements can add important insights into the roles of focal tissue damage and impaired local functionality in the pathogenesis of osteoarthritis.     

Contact models of repaired articular surfaces: influence of loading conditions and the superficial tangential zone

The superficial tangential zone (STZ) plays a significant role in normal articular cartilage’s ability to support loads and retain fluids. To date, tissue engineering efforts have not replicated normal STZ function in cartilage repairs. This finite element study examined the STZ’s role in normal and repaired articular surfaces under different contact conditions. Contact area and pressure distributions were allowed to change with time, tension-compression nonlinearity modeled collagen behavior in the STZ, and nonlinear geometry was incorporated to accommodate finite deformation. Responses to loading via impermeable and permeable rigid surfaces were compared to loading via normal cartilage, a more physiologic condition, anticipating the two rigid loading surfaces would bracket that of normal. For models loaded by normal cartilage, an STZ placed over the inferior repair region reduced the short-term axial compression of the articular surface by 15%, when compared to a repair without an STZ. Covering the repair with a normal STZ shifted the flow patterns and strain levels back toward that of normal cartilage. Additionally, reductions in von Mises stress (21%) and an increase in fluid pressure (13%) occurred in repair tissue under the STZ. This continues to show that STZ properties of sufficient quality are likely critical for the survival of transplanted constructs in vivo. However, response to loading via normal cartilage did not always fall within ranges predicted by the rigid surfaces. Use of more physiologic contact models is recommended for more accurate investigations into properties critical to the success of repair tissues.     

The role of chondrocyte-matrix interactions in maintaining and repairing articular cartilage

Throughout life chondrocytes maintain the articular cartilage matrix by replacing degraded macromolecules and respond to focal cartilage injury or degeneration by increasing local synthesis activity. These observations suggest that mechanisms exist within articular cartilage that stimulate chondrocyte anabolic activity in response to matrix degradation or damage. An important cartilage anabolic factor, insulin-like growth factor I (IGF-I), appears to have a role in stimulating chondrocyte anabolic activity. Although IGF-I is ubiquitous, its bioavailability is controlled by a class of secreted proteins, IGF binding proteins (IGFPBs). Of the six known IGFPBs, IGFBP-3 is the most abundant in human articular cartilage. We recently found that with increasing age, articular chondrocytes increase their expression of IGFBP-3. This observation led us to investigate the potential role of IGFBP-3 in chondrocyte-matrix interactions. Using immunofluorescent staining and confocal microscopy we found that IGFBP-3 accumulates with increasing age in the chondrocyte territorial matrix where it co-localizes with fibronectin, but not with tenascin-C or type VI collagen. Using purified proteins we demonstrated that IGFBP-3 binds to fibronectin in a dose dependent manner, but not to tenascin-C. In vitro studies showed that IGFBP-3 alone inhibited chondrocyte synthetic activity while intact fibronectin alone significantly stimulated activity. When fibronectin and IGFBP-3 were combined we found that the inhibitory activity of low concentrations of IGFPB-3 was enhanced. These observations indicate that in mature articular cartilage IGF-I is stored in the chondrocyte territorial matrix through binding to a complex of IGFPB-3 and intact fibronectin. Storage of IGF-I of the territorial matrix may help maintain a relatively constant level of available IGF-I and the local increase in matrix synthesis following matrix damage may result from release of IGF-I. This mechanism may have an important role in maintaining and repairing articular cartilage and failure of this mechanism may lead to progressive articular cartilage degeneration.     

Comparison of nonlinear mechanical properties of bovine articular cartilage and meniscus

Nonlinear, linear and failure properties of articular cartilage and meniscus in opposing contact surfaces are poorly known in tension. Relationships between the tensile properties of articular cartilage and meniscus in contact with each other within knee joints are also not known. In the present study, rectangular samples were prepared from the superficial lateral femoral condyle cartilage and lateral meniscus of bovine knee joints. Tensile tests were carried out with a loading rate of 5 mm/min until the tissue rupture. Nonlinear properties of the toe region, linear properties in larger strains, and failure properties of both tissues were analysed. The strain-dependent tensile modulus of the toe region, Young's modulus of the linear region, ultimate tensile stress and toughness were on average 98.2, 8.3, 4.0 and 1.9 times greater (p<0.05) for meniscus than for articular cartilage. In contrast, the toe region strain, yield strain and failure strain were on average 9.4, 3.1 and 2.3 times greater (p<0.05) for cartilage than for meniscus. There was a significant negative correlation between the strain-dependent tensile moduli of meniscus and articular cartilage samples within the same joints (r=−0.690, p=0.014). In conclusion, the meniscus possesses higher nonlinear and linear elastic stiffness and energy absorption capability before rupture than contacting articular cartilage, while cartilage has longer nonlinear region and can withstand greater strains before failure. These findings point out different load carrying demands that both articular cartilage and meniscus have to fulfil during normal physiological loading activities of knee joints.     

Load sharing between solid and fluid phases in articular cartilage: I--Experimental determination of in situ mechanical conditions in a porcine knee.

The in situ mechanical conditions of cartilage in the articulated knee were quantified during joint loading. Six porcine knees were subjected to a 445 N compressive load while cartilage deformations and contact pressures were measured. From roentgenograms, cartilage thickness before and during loading allowed the calculation of tissue deformation on the lateral femoral condyle at different times during the loading process. Contact pressures on the articular surface were measured with miniature fiber-optic pressure transducers. Results showed that the medial side of the lateral femoral condyle had higher contact pressures, as well as deformations. To begin to correlate the pressures and resulting deformations, the intrinsic material properties of the cartilage on the lateral condyle were obtained from indentation tests. Data from four normal control specimens indicated that the aggregate modulus of the medial side was significantly higher than in other areas of the condyle. These experimental measures of the in situ mechanical conditions of articular cartilage can be combined with theoretical modeling to obtain valuable information about the relative contributions of the solid and fluid phases to supporting the applied load on the cartilage surface (see Part II).     

Singular perturbation analysis of the nonlinear, flow-dependent compressive stress relaxation behavior of articular cartilage

The dominant mechanism giving rise to the viscoelastic response of articular cartilage during compression is the nonlinear diffusive interaction of the fluid and solid phases of the tissue as they flow relative to one another. The present study is concerned with the role of this interaction under uniaxial stress relaxation in compression. The model is a biphasic mixture of fluid and solid which incorporates the strain-dependent permeability found earlier from permeation experiments. When a ramp-displacement is imposed on the articular surface, simple, but accurate, asymptotic approximations are derived for the deformation and stress fields in the tissue for slow and moderately fast rates of compression. They are shown to agree very well with experiment and they provide a simple means for determining the material parameters. Moreover, they lead to important insights into the role of the flow-dependent viscoelastic nature of articular cartilage and other hydrated biological tissues.