The influence of link protein stabilization on the viscometric properties of proteoglycan aggregate solutions.

The dynamic, steady-shear and transient shear flow properties of precisely prepared link-stable (s0 136, 66% aggregate) and link-free (s0 93, 59% aggregate) proteoglycan aggregate solutions at concentrations ranging from 10 to 50 mg/ml were determined using a cone-on-plate viscometer in a mechanical spectrometer. All proteoglycan solutions tested possessed: (1) linear viscoelastic properties - as measured by the dynamic complex modulus under small amplitude steady oscillatory conditions (1 less than or equal to omega less than or equal to 100 rad/s) - and (2) nonlinear shear-rate dependent apparent viscosities and primary normal stress difference under steady shearing conditions (0.25 less than or equal to gamma less than or equal to 250 s-1). Our transient flow data show that all proteoglycan aggregate solutions exhibited transient stress overshoot effects in shear stress and normal stress. From these steady and transient flow data, we conclude that link protein stabilized aggregates have significant effects on their dynamic and steady-shear properties as well as transient flow properties. The transient stress overshoot data provide a measure of the energy per unit volume of fluid required to overcome the proteoglycan networks in solution from a resting state. Thus we found that link-stable aggregates form much stronger networks than link-free aggregates. This is corroborated by the fact that link-stable aggregates form more elastic (lower than delta) and stiffer (higher [G*]) networks than link-free aggregates. The complete spectrum of viscometric flow data is entirely compatible with the proposed role of link protein in adding structural stability to the proteoglycan-hyaluronate bond. In cartilage, the enhanced strength of the networks formed by link-stable aggregates may play an important role in determining the material properties of the tissue and thereby contribute to the functional capacity of cartilage in diarthrodial joints.

Cartilage and diarthrodial joints as paradigms for hierarchical materials and structures.

The anatomic forms of diarthrodial joints are important structural features which provide and limit the motions required for the joint. Typically, the length scale of topographic variation of anatomic forms ranges from 0.5 to 15 cm. Articular cartilage is the thin layer of hydrated soft tissue (0.5-5.0 mm thick) covering the articulating bony ends in diarthrodial joints. This tissue has a set of unique mechanical and physicochemical properties which are responsible for its load-carrying capabilities and near-frictionless qualities. The mechanical properties of articular cartilage are determined at the tissue-scale level and these properties depend on the composition of the tissue, mainly collagen and proteoglycan, and their molecular and ultrastructural organization (ultra-scale: 10(-8)-10(-6) m). Because proteoglycans possess a high density of fixed negative charges, articular cartilage exhibits a significant Donnan osmotic pressure effect. This physicochemically derived osmotic pressure is an important component of the total swelling pressure; the other component of the total swelling pressure stems from the charge-to-charge repulsive force exerted by the closely spaced (1-1.5 nm) negative charge groups along the proteoglycan molecules. Thus these interactions take place at a nano-scale level: 10(-10)-10(-9) m. Finally, cartilage biochemistry and organization are maintained by the chondrocytes which exist at a micro-scale level (10(-7)-10(-6) m). Significant mechanoelectrochemical transduction occurs within the extracellular matrix at the micro-scale level which affects and modulates cellular anabolic and catabolic activities. At present, the exact details of these transduction mechanisms are unknown. In this review, we present a summary of the hierarchical features for articular cartilage and diarthrodial joints and tables of known material properties for cartilage. Also we summarize how the multi-scale interactions in articular cartilage provide for its unique material properties and tribological characteristics.

Chondrocyte deformation and local tissue strain in articular cartilage: a confocal microscopy study.

It is well accepted that mechanical forces can modulate the metabolic activity of chondrocytes, although the specific mechanisms of mechanical signal transduction in articular cartilage are still unknown. One proposed pathway through which chondrocytes may perceive changes in their mechanical environment is directly through cellular deformation. An important step toward understanding the role of chondrocyte deformation in signal transduction is to determine the changes in the shape and volume of chondrocytes during applied compression of the tissue. Recently, a technique was developed for quantitative morphometry of viable chondrocytes within the extracellular matrix using three-dimensional confocal scanning laser microscopy. In the present study, this method was used to quantify changes in chondrocyte morphology and local tissue deformation in the surface, middle, and deep zones in explants of canine articular cartilage subjected to physiological levels of matrix deformation. The results indicated that at 15% surface-to-surface equilibrium strain in the tissue, a similar magnitude of local tissue strain occurs in the middle and deep zones. In the surface zone, local strains of 19% were observed, indicating that the compressive stiffness of the surface zone is significantly less than that of the middle and deep zones. With this degree of tissue deformation, significant decreases in cellular height of 26, 19, and 20% and in cell volume of 22, 16, and 17% were observed in the surface, middle, and deep zones, respectively. The deformation of chondrocytes in the surface zone was anisotropic, with significant lateral expansion occurring in the direction perpendicular to the local split-line pattern. When compression was removed, there was complete recovery of cellular morphology in all cases. These observations support the hypothesis that deformation of chondrocytes or a change in their volume may occur during in vivo joint loading and may have a role in the mechanical signal transduction pathway of articular cartilage.

Alterations in the mechanical behavior of the human lumbar nucleus pulposus with degeneration and aging.

This study tested the hypothesis that changes in the morphology and composition of the nucleus pulposus with age and degeneration have associated changes in its mechanical properties. A torsional shear experiment was used to determine viscoelastic shear properties of cylindrical samples of human nucleus pulposus with large ranges of grades of morphological degeneration (normal to severely degenerated) and ages (range: 16-88 years; average: 57 +/- 21.5 years). Viscoelastic shear properties were determined from stress-relaxation and dynamic sinusoidal tests. A linear viscoelastic law with a variable-amplitude relaxation spectrum was used to model experimental behaviors of nucleus pulposus specimens. A statistically significant increase in the instantaneous and dynamic shear moduli was found with increasing age and grade of degeneration; the values for moduli ranged from 5.0 to 60 kPa. A significant decrease in tan delta was also detected; the values ranged from 0.43 to 0.33, indicating a decreased capacity for the nucleus pulposus to dissipate energy. The dynamic modulus and tan delta were also significantly affected by frequency. It was generally concluded that the nucleus pulposus undergoes a transition from "fluid-like" behavior to more "solid-like" behavior with aging and degeneration.

Mechanical behavior of articular cartilage in shear is altered by transection of the anterior cruciate ligament.

The flow-independent viscoelastic and equilibrium behaviors of canine articular cartilage were examined with time after transection of the anterior cruciate ligament. The equilibrium, transient, and dynamic shear behaviors of cartilage were studied in biaxial compression-torsion testing at two time periods after transection of the anterior cruciate ligament and at two sites on the femoral condyle, in order to test for differences between sites of frequent and less frequent contact. Water content also was measured in cartilage at sites corresponding to the areas of mechanical testing. Transection of the anterior cruciate ligament produced significant decreases in all measured moduli of articular cartilage tested in equilibrium and dynamic shear and in equilibrium compression; the values for these moduli were 61, 56, and 77% of the control values, respectively, beginning at 6 weeks following transection of the anterior cruciate ligament. There was evidence of increased energy dissipation of cartilage in shear, with a 13 and 35% increase in tan delta at 6 and 12 weeks after transection of the anterior cruciate ligament, respectively. Changes in the viscoelastic relaxation function of cartilage in shear also were evident at 12 weeks after surgery. In all tissue, there was a significant increase in hydration of approximately 4% at 6 or 12 weeks after surgery. There was little difference between the material parameters for areas considered to be in frequent and less frequent contact, with the exception of hydration, which was greater for areas of less frequent contact.(ABSTRACT TRUNCATED AT 250 WORDS)

Radial tie fibers influence the tensile properties of the bovine medial meniscus.

Although collagen fibers are arranged predominantly in the circumferential direction in the knee meniscus, there is evidence for radially oriented fibers within human menisci. A bovine medial meniscus model was used to study the hypothesis that radial fibers alter the radial tensile properties of the meniscus. The architecture of the collagen network and tensile properties of the bovine medial meniscus were examined; attention was given to large "radial tie fibers" and their regional variation. Menisci were sectioned serially into slices 400 microns thick. Polarized light microscopy showed that the distribution of radial tie fibers varied greatly among the anterior, central, and posterior regions. These radial tie fibers were larger and more frequent in the posterior region. Radial fibers persisted over many adjacent sections with similar architecture, which led to our hypothesis that they may be arranged in continuous sheets in which the morphology varies by region. Radially oriented specimens for tensile testing were grouped according to the number of radial tie fibers (full, partial, and no fiber) and region (anterior, central, and posterior). Uniaxial tensile testing was performed on a testing machine at a strain rate of 0.00017 sec-1 until failure. The tensile modulus, ultimate tensile stress, and ultimate tensile strain were determined. The presence of radial tie fibers in the specimen had a significant effect on the tensile modulus and ultimate tensile stress. Specimens containing full radial tie fibers were stiffest and failed at the lowest strains; in specimens from the posterior region, the tensile modulus was 392%, the ultimate tensile stress was 314%, and the ultimate tensile strain was 68% that of the specimens with no radial fibers. In no-fiber specimens, the tensile modulus in the posterior region was 225% of the modulus in the anterior region, and the ultimate tensile strain in the posterior region was 68% that of the strain in the anterior region. The abundance of radial tie fibers in the posterior region seems to contribute to the increased stiffness of this region. The preferential stiffening of the posterior region by these radial fibrous sheets may be well suited to the manner in which the bovine medial meniscus functions in load-bearing.

Viscoelastic shear properties of articular cartilage and the effects of glycosidase treatments.

The objectives of this study were to determine the viscoelastic shear properties of articular cartilage and to investigate the effects of the alteration of proteoglycan structure on these shear properties. Glycosidase treatments (chondroitinase ABC and Streptomyces hyaluronidase) were used to alter the proteoglycan structure and content of the tissue. The dynamic viscoelastic shear properties of control and treated tissues were measured and statistically compared. Specifically, cylindrical bovine cartilage specimens were subjected to oscillatory shear deformation of small amplitude (gamma degrees = 0.001 radian) over a physiological range of frequencies (0.01-20 Hz) and at various compressive strains (5, 9, 12, and 16%). The dynamic complex shear modulus was calculated from the measurements. The experimental results show that the solid matrix of normal articular cartilage exhibits intrinsic viscoelastic properties in shear over the range of frequencies tested. These viscoelastic shear properties were found to be dependent on compressive strains. Our data also provide significant insights into the structure-function relationships for articular cartilage. Significant correlations were found between the material properties (the magnitude of dynamic shear modulus, the phase shift angle, and the equilibrium compressive modulus), and the biochemical compositions of the cartilage (collagen, proteoglycan, and water contents). The shear modulus was greatly reduced when the proteoglycans were degraded by either chondroitinase ABC or Streptomyces hyaluronidase. The results suggest that the ability of collagen to resist tension elastically provides the stiffness of the cartilage matrix in shear and its elastic energy storage capability. Proteoglycans enmeshed in the collagen matrix inflate the collagen network and induce a tensile prestress in the collagen fibrils. This interaction of the collagen and proteoglycan within the cartilage matrix provides the complex mechanism that allows the tissue to resist shear deformation.

Viscoelastic properties of proteoglycan solutions with varying proportions present as aggregates.

Monomer and aggregated proteoglycans were prepared from pig laryngeal cartilage. Vascoelastic flow properties, comprising linear complex dynamic shear modulus, nonlinear steady-state shear-rate dependent viscosity, and primary normal stress difference, were measured in proteoglycan solutions containing varying proportions of aggregate (0-80%) and at different concentrations (10-50 mg/ml). Results were analyzed using the simple Oldroyd four-parameter nonlinear rate-type rheological equation. All solution properties were strongly dependent on proteoglycan concentration and on the proportion of aggregates present. Aggregation was found to have a great effect on the zero shear-rate viscosity at 50 mg/ml, which increased fivefold from 0-100% aggregate. The results showed that network formation in proteoglycan solutions increased with concentration from 10-50 mg/ml and also increased with aggregation. All proteoglycan solutions showed shear thinning, which was most marked with aggregated proteoglycan at high concentration (50 mg/ml), where the viscosity decreased tenfold from the zero shear-rate limit to the infinite shear-rate limit. The intermolecular interactions in the network were therefore increasingly disrupted by increasing shear rate, but repeated measurements showed that these were reversible changes and that testing did not induce disaggregation or degradation of proteoglycan. These rheological properties show that aggregation is likely to immobilize proteoglycan at high concentration within cartilage and to contribute to the material properties of the porous solid matrix of articular cartilage that are important for its load-bearing function.

Tensile properties of human knee joint cartilage: I. Influence of ionic conditions, weight bearing, and fibrillation on the tensile modulus.

The flow-independent (intrinsic) tensile modulus of the extracellular matrix of human knee joint cartilage has been measured for normal, fibrillated, and osteoarthritic (removed from total knee joint replacements) cartilage. The modulus was determined in our isometric tensile apparatus and measured at equilibrium. We found a linear equilibrium stress-strain behavior up to approximately 15% strain. The modulus was measured for tissues from the high and low weight-bearing areas of the joint surfaces, the medial femoral condyle and lateral patello femoral groove, and from different zones (surface, subsurface, middle, and middle-deep) within the tissue. For all specimens, the intrinsic tensile modulus was always less than 30 MPa. Tissues from low weight-bearing areas (LWA) are stiffer than those from high weight-bearing areas (HWA). The tensile modulus of the ECM correlates strongly with the collagen/proteoglycan ratio; it is higher for LWA than for HWA. Osteoarthritic cartilage from total knee replacement procedures has a tensile stiffness less than 2 MPa.

Tensile properties of human knee joint cartilage. II. Correlations between weight bearing and tissue pathology and the kinetics of swelling.

The nonequilibrium or kinetic swelling behavior of normal, fibrillated, and osteoarthritic (OA) (removed from total knee joint replacements) human knee joint cartilage has been measured using our isometric tensile apparatus (ITA). We found that large local variations exist in the manner with which human knee joint cartilage swells, including anisotropic effects, inhomogeneities, and dependence on local biochemical composition and pathological condition. The ITA provides three convenient biomechanical parameters--peak stress (sigma p), stress relaxation (sigma R), and diffusion coefficient (D)--to quantify the kinetics of swelling. We used these parameters to quantify and differentiate the kinetic swelling behavior of normal, fibrillated, and osteoarthritic cartilage, as well as the swelling behavior of cartilage from high and low weight-bearing areas. Also, these kinetic swelling parameters correlated very well, though by varying degrees, with such biochemical measures as collagen/proteoglycan ratio, hexosamine content/wet weight, and hydroxyproline content/dry weight, providing important insight into the mechanisms and processes involved during the course of swelling. Hence, the kinetic swelling behavior of cartilage should be used to provide important information not obtainable from equilibrium swelling studies.

The effect of cryopreservation on canine menisci: a biochemical, morphologic, and biomechanical evaluation.

This study evaluated the effect of cryopreservation on the structural organization, biosynthetic activity, and material properties of canine menisci. The menisci were cryopreserved by incubating them in a 4% solution of dimethyl sulfoxide (DMSO) in physiologic media and freezing them to -100 degrees C using a controlled rate freezing system. The menisci were then stored for varying periods of time from zero to 12 weeks in liquid nitrogen (-196 degrees C). Following rapid thawing, changes in the histological appearance and biosynthetic activity of the menisci were evaluated as functions of storage time. In addition, the effects of the cryopreservation process on the tensile strength and modulus of the meniscal tissue were assessed. Although cryopreservation and short-term storage did not appear to affect the morphological appearance or biomechanical character of the menisci, biosynthetic activity, as determined by Na2S35SO4 incorporation, was diminished to less than 50% of normal control values immediately following cryopreservation and thawing. Autoradiographic examination of these tissues revealed that only approximately 10% of the meniscal cells were metabolically active, however, indicating that a marked increase in the metabolic activity of individual cells occurs following the freeze-thaw cycle. Total metabolic activity continued to decline with storage time.

Material properties of the normal medial bovine meniscus.

The intrinsic compressive and tensile properties of normal bovine medial menisci were measured, and the variations in these properties with respect to the structural organization of the tissue and test specimen location were examined. Using a confined compression experiment, the compressive aggregate modulus and permeability of the meniscus were determined. The permeability of the tissue was also compared with the permeability as measured experimentally using a direct permeation experiment. Deep posterior specimens are significantly stiffer in compression than deep anterior and central-anterior specimens, while deep anterior specimens are significantly less stiff than deep posterior and central-posterior specimens. Posterior specimens have a significantly higher average water content. In addition, the permeability of the bovine meniscus was found to be about one-tenth that of bovine articular cartilage. The tensile stiffness of meniscal tissue was determined from constant strain rate uniaxial tension tests. To asses the directional variations in the tensile properties, specimens were obtained from the circumferential and radial orientations. The results indicate that the femoral surface of the meniscus is isotropic in tension, while specimens from within the meniscus are anisotropic--the circumferential specimens are much stiffer than the radial specimens. Furthermore, circumferential posterior specimens from the interior of the meniscus are significantly stiffer than similar anterior specimens. Layer inhomogeneities in the tensile properties with respect to distance from the femoral surface are also present.

Effects of proteoglycan extraction on the tensile behavior of articular cartilage.

We undertook an interdisciplinary biomechanical and biochemical study to explore the extent and manner in which the total pool of proteoglycans influences the kinetic and static behavior of bovine articular cartilage in tension. Two biomechanical tests were used: (a) the viscoelastic creep test and (b) a slow constant-rate uniaxial tension test; and two enzymatic proteoglycan extraction procedures were used: (a) chondroitinase ABC treatment and (b) a sequential enzymatic treatment with chondroitinase ABC, trypsin, and Streptomyces hyaluronidase. We found that the viscoelastic creep response of all cartilage specimens may be divided into two distinct phases: an initial phase (less than 15 s), characterized by a rapid increase in strain following load application, and a late phase (15 s less than or equal to t less than 25,000 s), characterized by a more gradual increase in strain. A major finding of this study is that the kinetics of the creep response is greatly influenced by the glycosaminoglycan content of the tissue. For untreated and control specimens, the initial response comprises about 50% of the total strain, while for chondroitinase ABC and sequentially extracted specimens, the initial response comprises up to 83% of the total strain. Furthermore, most untreated and control specimens did not reach equilibrium within the 25,000 s test period, while enzymatically digested specimens often reached equilibrium in less than 100 s. Thus, we conclude that through their physical restraints on collagen, the bulk of proteoglycan present in the tissue acts to retard fibrillar reorganization and alignment under tensile loading, thereby effectively preventing sudden extension of the collagen network. In contrast, the results of our slow constant-rate uniaxial tension experiment show that essentially complete extraction of proteoglycan glycosaminoglycans does not affect the intrinsic tensile stiffness and strength of cartilage specimens or the collagen network in a significant manner. Hence, an important function of the bulk proteoglycans (i.e., the large aggregating type) in cartilage is to retard the rate of stretch and alignment when a tensile load is suddenly applied. This mechanism may be useful in protecting the cartilage collagen network during physiological situations, where sudden impact forces are imposed on a joint.

Compressive stress-relaxation behavior of bovine growth plate may be described by the nonlinear biphasic theory.

The compressive behavior of the bovine distal femoral growth plate was studied in vitro. Strain-rate controlled, compression stress-relaxation experiments were performed on cylindrical bone-growth plate-bone specimens from the interior and periphery of the growth plate. The questions addressed in this study were (a) Can the nonlinear biphasic theory, one with strain-dependent permeability, be used to represent the compressive stress-relaxation behavior of bovine growth plate? (b) How do different assumptions concerning the permeabilities of the chondro-osseous interfaces influence the inferred material properties of the growth plate? and (c) Are there any differences in these properties between the periphery and the interior of the growth plate? Intrinsic biphasic material properties--aggregate modulus (HA), Poisson's ratio (v), and nonlinear strain-dependent permeability coefficients (ko and M)--were calculated from the compression stress-relaxation data with use of a finite element model and a least squares curve-fitting procedure. To verify this constitutive model for the growth plate, an independent set of finite element analyses was performed with use of the determined intrinsic biphasic properties, and comparisons were made between these finite element predictions and two additional sets of experimental data subsequently obtained for the same specimens with use of two slower rates of compression. Excellent agreement was achieved between these finite element predictions and the latter two sets of data. The aggregate modulus was found to be insensitive to the permeability of the chondro-osseous interface. The permeability coefficients were very sensitive to, and the Poisson's ratio was only slightly sensitive to the interface permeability condition. Therefore, the periphery of the growth plate is more compliant and permeable than the interior.

Mechanical properties of canine articular cartilage are significantly altered following transection of the anterior cruciate ligament.

The compressive, tensile, and swelling properties of articular cartilage were studied at two time periods following transection of the anterior cruciate ligament in the knee of greyhound dogs. An experimental protocol was designed to quantify the essential equilibrium and biphasic material properties of cartilage in tension, compression, and shear, as well as the parameters of isometric swelling behavior. All properties were measured at several sites to elicit differences between sites of frequent and less frequent contact. Hydration was determined at each site and was compared with the material properties of cartilage from corresponding sites. There were extensive changes in all compressive, tensile, and swelling properties of cartilage after transection of the anterior cruciate ligament. Twelve weeks after surgery, the intrinsic moduli were reduced significantly in compression (approximately 24% of control values), tension (approximately 64%), and shear (approximately 24%), and the hydraulic permeability was elevated significantly (approximately 48%). Significant increases in hydration (approximately 9%) also were observed, as well as a strong correlation of hydration with hydraulic permeability. The pattern of these changes was not found to differ with site in the joint, but significant differences were observed in the magnitude of change for cartilage from the femoral groove and the femoral condyle. The pattern and extent of changes in the material properties following transection of the anterior cruciate ligament indicate that altered loading of the joint severely compromises the overall mechanical behavior of articular cartilage. The observed loss of matrix stiffness in compression, tension, and shear is associated with increases in the deformation of the solid matrix, a diminished ability to resist swelling, and the increase in hydration observed in this study. The increased swelling and elevated water content were related directly to the increase in hydraulic permeability; this suggests an associated loss of fluid pressurization as the load support mechanism in the degenerated cartilage. Without a successful mechanism for repair, damage to the solid matrix may progress and lead to further degenerative changes in the biochemistry, morphology, and mechanical behavior of articular cartilage.

Mechanical and biochemical changes in the superficial zone of articular cartilage in canine experimental osteoarthritis.

The changes in the tensile mechanical properties and biochemical composition of the superficial zone of articular cartilage were examined in a canine model of early osteoarthritis generated by transection of the anterior cruciate ligament. Sixteen weeks following ligament transection, the tensile stiffness of the articular cartilage was decreased by 44% and the ion-induced stress relaxation of the tissue was increased by 57% compared with the contralateral control. Biochemical analyses indicated that the water content of the experimental tissue was increased by 13%, which was reflected as an apparent 37% decrease in the proteoglycan content and a 36% decrease in the collagen content (expressed per wet weight). The hydroxypyridinium crosslink density was decreased in the experimental tissue by 11%. A significant negative correlation was found between the ion-induced stress relaxation and the hydroxypyridinium crosslink density in both control tissue (R = -0.56) and experimental tissue (R = -0.70). No correlation was noted between the tensile stiffness and the biochemical composition of the tissue. These results suggest that, in the superficial zone of articular cartilage, the structure of the tissue may play a more important role than the composition in the determination of its mechanical properties. A major event observed in the model of early osteoarthritis appears to be the disruption and remodeling of the collagen network in the superficial zone of the articular cartilage.

Interspecies comparisons of in situ intrinsic mechanical properties of distal femoral cartilage.

We measured the in situ biomechanical properties of knee joint cartilage from five species (bovine, canine, human, monkey, and rabbit) to examine the biomechanical relevance of animal models of human knee joint injuries and osteoarthritis. In situ biphasic creep indentation experiments were performed to simultaneously determine all three intrinsic material coefficients (aggregate modulus, Poisson's ratio, and permeability) of the cartilage as represented by the linear KLM biphasic model. In addition, we also assessed the effects of load bearing on these intrinsic properties at "high" and "low" weight-bearing regions on the distal femur. Our results indicate that significant differences exist in some of these material properties among species and sites. The aggregate modulus of the anterior patellar groove within each species is the lowest among all sites tested, and the permeability of the patellar groove cartilage is the highest and does not vary among species. Similarly, the Poison's ratio in the patellar groove is the lowest in all species, except in the rabbit. These results lead to the conclusion that patellar groove cartilage can undergo greater and faster compression. Thus, under high compressive loads, the cartilage of the patellar groove surface can more rapidly compress to create a congruent patellofemoral joint articulation. For any given location, no differences were found in the aggregate modulus among all the species, and no correlation was found between aggregate modulus and thickness at the test site. Thus, in the process of selecting a suitable experimental animal model of human articular cartilage, it is essential to consider the significant interspecies differences of the mechanical properties.

In vivo effects of naproxen on composition, proteoglycan metabolism, and matrix metalloproteinase activities in canine articular cartilage.

Naproxen is a nonsteroidal anti-inflammatory drug commonly used in the clinical treatment of joint disease. In this study, its effect in vivo on the biochemical composition, metabolic activities, and metalloproteinase activities of normal canine articular cartilage was analyzed. The articular cartilage from the knee joints of dogs who had been given naproxen for 4 weeks to maintain a serum level of 40-50 micrograms/ml was examined. Control animals were given a placebo. Treatment with naproxen was not found to change the composition (water, collagen, and proteoglycan) of the articular cartilage. The culture studies of cartilage explants indicated that proteoglycan synthesis rates were unaffected by the treatment with naproxen but that proteoglycan release from the tissue was suppressed. Analysis of the cartilage for matrix metalloproteinase activities showed reduced activity of neutral matrix metalloproteinase by 80%, of collagenase by 40%, and of gelatinase by 87%, with no change in activity of acid metalloproteinase or of tissue inhibitor for metalloproteinase. These findings indicate that in vivo treatment with naproxen has the capacity to modulate catabolic activities in articular cartilage.

Compressive properties of the cartilaginous end-plate of the baboon lumbar spine.

The viscoelastic behavior of the cartilaginous end-plate of the baboon (Papio anubis) was studied in an experiment on compressive creep. Data were analyzed with the biphasic poroviscoelastic constitutive theory to assess the relative contributions of flow-dependent and flow-independent viscoelastic mechanisms to the observed creep behavior. Material coefficients describing the equilibrium compressive behavior (HA) and both flow-independent (c, tau 1, and tau 2) and flow-dependent (k) viscoelastic effects were determined for the end-plate by the curve-fitting of the theoretical solution to the experimental creep data. Biochemical analyses were performed to test for potential relationships between material properties and composition which may give rise to the viscoelastic behavior of the end-plate. The results indicate that the cartilaginous end-plate has a hydraulic permeability of 14.3 x 10(-14) m4/N-s, which is associated with rapid transport and pressurization of the interstitial fluid in response to loading and an increased emphasis on flow-independent viscoelastic effects. Biochemical analyses for water, sulfated glycosaminoglycan content, and hydroxyproline indicate that the end-plate of the baboon is compositionally similar to the cartilaginous end-plate in humans. Interpretation of the mechanical and compositional data suggests that fluid pressurization in the cartilaginous end-plate may be important in the maintenance of a uniform stress distribution across the boundary between vertebral body and intervertebral disc.

Shear mechanical properties of human lumbar annulus fibrosus.

Function, failure, and remodeling of the intervertebral disc are all related to the stress and strain fields in the tissue and may be calculated by finite element models with accurate material properties, realistic geometry, and appropriate boundary conditions. There is no comprehensive study in the literature investigating the shear material properties of the annulus fibrosus. This study obtained shear material properties of the annulus fibrosus and tested the hypothesis that these properties are affected by the amplitude and frequency of shearing, applied compressive stress, and degenerative state of the tissue. Cylindrical specimens with an axial orientation from seven nondegenerated and six degenerated discs were tested in torsional shear under dynamic and static conditions. Frequency sweep experiments over a physiological range of frequencies (0.1-100 rad/sec) at a shear strain amplitude of 0.05 rad were performed under three different axial compressive stresses (17.5, 25, and 35 kPa). At the largest compressive stress, shear strain sweep experiments (strain amplitude range: 0.005-0.15 rad at a frequency of 5 rad/sec) and transient stress-relaxation tests (shear strain range: 0.02-0.15 rad) were performed. The annulus fibrosus material was less stiff and more dissipative at larger shear strain amplitudes, stiffer at higher frequencies of oscillation, and stiffer and less dissipative at larger axial compressive stresses. The dynamic shear modulus, /G*/, had values ranging from 100 to 400 kPa, depending on the experimental condition and degenerative level. The shear behavior was also predominantly elastic, with values for the tangent of the phase angle (tandelta) ranging from 0.1 to 0.7. The annulus material also became stiffer and more dissipative with degenerative grade; however, this was not statistically significant. The results indicated that nonlinearities, compression/shear coupling, intrinsic viscoelasticity, and, to a lesser degree, degeneration all affect the shear material behavior of the annulus fibrosus, with important implications for load-carriage mechanisms in the intervertebral disc. These material complexities should be considered when choosing material constants for finite element models.