Immature bovine cartilage wear is due to fatigue failure from repetitive compressive forces and not reciprocating frictional forces.

OBJECTIVE: Wear of articular cartilage is not well understood. We hypothesize that cartilage wears due to fatigue failure in repetitive compression instead of reciprocating friction. DESIGN: This study compares reciprocating sliding of immature bovine articular cartilage against glass in two testing configurations: (1) a stationary contact area configuration (SCA), which results in static compression, interstitial fluid depressurization, and increasing friction coefficient during reciprocating sliding, and (2) a migrating contact area configuration (MCA), which maintains pressurization and low friction while producing repetitive compressive loading in addition to reciprocating sliding. Contact pressure, sliding duration, and sliding distance were controlled to be similar between test groups. RESULTS: SCA tests exhibited an average friction coefficient of µ=0.084±0.032, while MCA tests exhibited a lower average friction coefficient of µ=0.020±0.008 (p<10-4). Despite the lower friction, MCA cartilage samples exhibited clear surface damage with a significantly greater average surface deviation from a fitted plane after wear testing (Rq=0.125±0.095 mm) than cartilage samples slid in a SCA configuration (Rq=0.044±0.017 mm, p=0.002), which showed minimal signs of wear. Polarized light microscopy confirmed that delamination damage occurred between the superficial and middle zones of the articular cartilage in MCA samples. CONCLUSIONS: The greatest wear was observed in the group with lowest friction coefficient, subjected to cyclical instead of static compression, implying that friction is not the primary driver of cartilage wear. Delamination between superficial and middle zones implies the main mode of wear is fatigue failure under cyclical compression, not fatigue or abrasion due to reciprocating frictional sliding.

Toward understanding the role of cartilage particulates in synovial inflammation.

OBJECTIVE: Arthroscopy with lavage and synovectomy can remove tissue debris from the joint space and the synovial lining to provide pain relief to patients with osteoarthritis (OA). Here, we developed an in vitro model to study the interaction of cartilage wear particles with fibroblast-like synoviocytes (FLS) to better understand the interplay of cartilage particulates with cytokines on cells of the synovium. METHOD: In this study sub-10 µm cartilage particles or 1 µm latex particles were co-cultured with FLS ±10 ng/mL interleukin-1α (IL-1α) or tumor necrosis factor-α (TNF-α). Samples were analyzed for DNA, glycosaminoglycan (GAG), and collagen, and media samples were analyzed for media GAG, nitric oxide (NO) and prostaglandin-E2 (PGE2). The nature of the physical interaction between the particles and FLS was determined by microscopy. RESULTS: Both latex and cartilage particles could be phagocytosed by FLS. Cartilage particles were internalized and attached to the surface of both dense monolayers and individual cells. Co-culture of FLS with cartilage particulates resulted in a significant increase in cell sheet DNA and collagen content as well as NO and PGE2 synthesis compared to control and latex treated groups. CONCLUSION: The proliferative response of FLS to cartilage wear particles resulted in an overall increase in extracellular matrix (ECM) content, analogous to the thickening of the synovial lining observed in OA patients. Understanding how cartilage particles interface with the synovium may provide insight into how this interaction contributes to OA progression and may guide the role of lavage and synovectomy for degenerative disease.

Passage-dependent relationship between mesenchymal stem cell mobilization and chondrogenic potential.

OBJECTIVE: Galvanotaxis, the migratory response of cells in response to electrical stimulation, has been implicated in development and wound healing. The use of mesenchymal stem cells (MSCs) from the synovium (synovium-derived stem cells, SDSCs) has been investigated for repair strategies. Expansion of SDSCs is necessary to achieve clinically relevant cell numbers; however, the effects of culture passage on their subsequent cartilaginous extracellular matrix production are not well understood. METHODS: Over four passages of SDSCs, we measured the expression of cell surface markers (CD31, CD34, CD49c, CD73) and assessed their migratory potential in response to applied direct current (DC) electric field. Cells from each passage were also used to form micropellets to assess the degree of cartilage-like tissue formation. RESULTS: Expression of CD31, CD34, and CD49c remained constant throughout cell expansion; CD73 showed a transient increase through the first two passages. Correspondingly, we observed that early passage SDSCs exhibit anodal migration when subjected to applied DC electric field strength of 6 V/cm. By passage 3, CD73 expression significantly decreased; these cells exhibited cell migration toward the cathode, as previously observed for terminally differentiated chondrocytes. Only late passage cells (P4) were capable of developing cartilage-like tissue in micropellet culture. CONCLUSIONS: Our results show cell priming protocols carried out for four passages selectively differentiate stem cells to behave like chondrocytes, both in their motility response to applied electric field and their production of cartilaginous tissue.

Time and dose-dependent effects of chondroitinase ABC on growth of engineered cartilage.

Tissue engineering techniques have been effective in developing cartilage-like tissues in vitro. However, many scaffold-based approaches to cultivating engineered cartilage have been limited by low collagen production, an impediment for attaining native functional load-bearing tensile mechanical properties. Enzymatic digestion of glycosaminoglycans (GAG) with chondroitinase ABC (chABC) temporarily suppresses the construct's GAG content and compressive modulus and increases collagen content. Based on the promising results of these early studies, the aim of this study was to further promote collagen deposition through more frequent chABC treatments. Weekly dosing of chABC at a concentration of 0.15 U/mL resulted in a significant cell death, which impacted the ability of the engineered cartilage to fully recover GAG and compressive mechanical properties. In light of these findings, the influence of lower chABC dosage on engineered tissue (0.004 and 0.015 U/mL) over a longer duration (one week) was investigated. Treatment with 0.004 U/mL reduced cell death, decreased the recovery time needed to achieve native compressive mechanical properties and GAG content, and resulted in a collagen content that was 65 % greater than the control. In conclusion, the results of this study demonstrate that longer chABC treatment (one week) at low concentrations can be used to improve collagen content in developing engineered cartilage more expediently than standard chABC treatments of higher chABC doses administered over brief durations.

Shearing of synovial fluid activates latent TGF-ß.

OBJECTIVE: TGF-ß is synthesized in an inactive latent complex that is unable to bind to membrane receptors, thus unable to induce a cellular biological response until it has been activated. In addition to activation by chemical mediators, recent studies have demonstrated that mechanical forces may activate latent TGF-ßvia integrin-mediated cellular contractions, or mechanical shearing of blood serum. Since TGF-ß is present in synovial fluid in latent form, and since normal diarthrodial joint function produces fluid shear, this study tested the hypothesis that the native latent TGF-ß1 of synovial fluid can be activated by shearing. DESIGN: Synovial fluid from 26 bovine joints and three adult human joints was sheared at mean shear rates up to 4000 s(-1) for up to 15 h. RESULTS: Unsheared synovial fluid was found to contain high levels of latent TGF-ß1 (4.35 ± 2.02 ng/mL bovine, 1.84 ± 0.89 ng/mL human; mean ± radius of 95% confidence interval) and low amounts (<0.05 ng/mL) of the active peptide. Synovial fluid concentrations of active TGF-ß1 increased monotonically with shear rate and shearing duration, reaching levels of 2.64 ± 1.22 ng/mL for bovine and 0.60 ± 0.39 ng/mL for human synovial fluid. Following termination of shearing, there was no statistical change in these active levels over the next 8 h for either species, demonstrating long-term stability of the activated peptide. The unsheared control group continued to exhibit negligible levels of active TGF-ß1 at all times. CONCLUSIONS: Results confirmed the hypothesis of this study and suggest that shearing of synovial fluid might contribute an additional biosynthetic effect of mechanical loading of diarthrodial joints.

Continuum mixture models of biological growth and remodeling: past successes and future opportunities.

Biological growth processes involve mass exchanges that increase, decrease, or replace material that constitutes cells, tissues, and organs. In most cases, such exchanges alter the structural makeup of the material and consequently affect associated mechanobiological responses to applied loads. Given that the type and extent of changes in structural integrity depend on the different constituents involved (e.g., particular cytoskeletal or extracellular matrix proteins), the continuum theory of mixtures is ideally suited to model the mechanics of growth and remodeling. The goal of this review is twofold: first, to highlight a few illustrative examples that show diverse applications of mixture theory to describe biological growth and/or remodeling; second, to identify some open problems in the fields of modeling soft-tissue growth and remodeling.

Perspectives on biological growth and remodeling.

The continuum mechanical treatment of biological growth and remodeling has attracted considerable attention over the past fifteen years. Many aspects of these problems are now well-understood, yet there remain areas in need of significant development from the standpoint of experiments, theory, and computation. In this perspective paper we review the state of the field and highlight open questions, challenges, and avenues for further development.

Response of engineered cartilage to mechanical insult depends on construct maturity.

UNLABELLED: Injury to articular cartilage leads to degenerative changes resulting in a loss of mechanical and biochemical properties. In engineered cartilage, the injury response of developing constructs is unclear. OBJECTIVE: To characterize the cellular response of tissue-engineered constructs cultured in chemically-defined medium after mechanical insult, either by compression-induced cracking, or by cutting, as a function of construct maturity. METHODS: Primary immature bovine articular chondrocytes (4-6 weeks) were encapsulated in agarose hydrogel (2%, 30 millioncells/mL) and cultured in chemically-defined medium supplemented with Transforming growth factor (TGF)-ß3 (10ng/mL, first 2 weeks). At early (5 days) and late (35 days) times in culture, subsets of constructs were exposed to mechanical overload to produce a crack in the tissue or were exposed to a sharp wound with a perpendicular cut. Constructs were returned to culture and allowed to recover in static conditions. Mechanical and biochemical properties were evaluated at 2-week intervals to day 70, and cellular viability was assessed at 2-week intervals to day 85. RESULTS: Constructs injured early in culture recovered their mechanical stiffness back to control values, regardless of the mode of injury. Later in culture, when constructs exhibited properties similar to those of native cartilage, compression-induced cracking catastrophically damaged the bulk matrix of the tissue and resulted in permanent mechanical failure with persistent cell death. No such detrimental outcomes were observed with cutting. Biochemical content was similar across all groups irrespective of mode or time of injury. CONCLUSIONS: Unlike native cartilage, engineered cartilage constructs exhibit a reparative capacity when the bulk integrity of the developing tissue is preserved after injury.

Mechanical properties of bovine articular cartilage under microscale indentation loading from atomic force microscopy.

Atomic force microscopy (AFM) techniques have been increasingly used for investigating the mechanical properties of articular cartilage. According to the previous studies reporting the microscale Young's modulus under AFM indentation tests, the Hertz contact model has been employed with a sharp conical tip indenter. However, the non-linear microscale behaviour of articular cartilage could not be resolved by the standardized Hertz analysis using small and sharp atomic force microscope tips. Therefore, the objective of this study was to evaluate the microscale Young's modulus of articular cartilage more accurately through a non-Hertzian approach with a spherical tip of 5 microm diameter, and to characterize its microscale mechanical behaviour. This methodology adopted in the present study was proved by the consistent values between the microscale (2 per cent, about 9.3 kPa; 3 per cent, about 17.5kPa) and macroscale (2 per cent, about 8.3kPa; 3 per cent, about 18.3kPa) Young's moduli for 2 per cent and 3 per cent agarose gel (n = 100). Therefore, the microscale Young's modulus evaluated in this study is representative of more accurate measurements of cartilage stiffness at the 600 nm deformation level and corresponds to approximately 30.9 kPa (n = 100). Furthermore, on this level of the microscale deformation, articular cartilage showed depth-dependent and frequency-independent behaviour under AFM indentation loading. These findings reveal the microscale mechanical behaviour of articular cartilage more accurately and can be employed further to design microscale structures of chondrocyte-seeded scaffolds and tissue-engineered cartilage by evaluating their microscale properties.

Investigation of the frictional response of osteoarthritic human tibiofemoral joints and the potential beneficial tribological effect of healthy synovial fluid.

OBJECTIVE: This study tests the hypothesis that the natural progression of osteoarthritis (OA) in human joints leads to an increase in the friction coefficient. This hypothesis is based on the expectation that the wear observed in OA may be exacerbated by higher friction coefficients. A corollary hypothesis is that healthy synovial fluid (SF) may help mitigate the increase in the friction coefficient in diseased joints. DESIGN: The friction coefficient of human tibiofemoral joints with varying degrees of OA was measured in healthy bovine SF and physiological buffered saline (PBS). Two testing configurations were adopted, one that promotes sustained cartilage interstitial fluid pressurization to investigate the effectiveness of this mechanism with advancing OA, and another that allows interstitial fluid pressure to subside to investigate the effectiveness of boundary lubrication. RESULTS: Seven specimens were visually staged to be normal or mildly degenerated (stages< or =2 on a scale of 1 to 4) and nine others had progressive degeneration (stages>2 and< or =3). No statistical differences were found in the friction coefficient with increasing OA, whether in migrating or stationary contact area configurations; however, the friction coefficient was significantly lower in SF than PBS in both configurations. CONCLUSIONS: The friction coefficient of human tibiofemoral cartilage does not necessarily increase with naturally increasing OA, for visual stages ranging from 1 to 3. This outcome may be explained by the fact that interstitial fluid pressurization is not necessarily defeated by advancing degeneration. This study also demonstrates that healthy SF decreases the friction coefficient of OA joints relative to PBS.

Scaffold degradation elevates the collagen content and dynamic compressive modulus in engineered articular cartilage.

OBJECTIVE: It was hypothesized that controlled, scaffold removal in engineered cartilage constructs would improve their collagen content and mechanical properties over time in culture. DESIGN: Preliminary experiments characterized the effects of agarase on cell-free agarose disks and cartilage explants. Immature bovine chondrocytes were encapsulated in agarose, cultured to day 42, and incubated with 100 units/mL agarase for 48 h. After treatment, constructs were cultured to day 91. The compressive Young's modulus and dynamic modulus of the constructs were determined every 2 weeks and immediately after agarase treatment. Post-mechanical testing, constructs were processed for biochemistry and histology. RESULTS: Agarase treatment on explants had no detrimental effect on the cartilage matrix. Treatment applied to engineered constructs on day 42 did not affect DNA or collagen content. Agarase treatment decreased tissue GAG content (via GAG loss to the media) and Young's modulus, both of which recovered to control values over time in culture. By day 91 agarase-treated constructs possessed approximately 25% more DNA, approximately 60% more collagen, and approximately 40% higher dynamic modulus compared to untreated controls. CONCLUSIONS: Scaffold degradation increased construct collagen content and dynamic mechanical properties, affirming the experimental hypothesis. The mechanism may lie in increased nutrient transport, increased space for collagen fibril formation, and cellular response to the loss of GAG with agarase treatment. The results highlight the role of the scaffold in retaining synthesized matrix during early and late tissue formation. This work also shows promise in developing an engineered tissue that may be completely free of scaffold material for clinical implantation.

Analysis of radial variations in material properties and matrix composition of chondrocyte-seeded agarose hydrogel constructs.

OBJECTIVE: To examine the radial variations in engineered cartilage that may result due to radial fluid flow during dynamic compressive loading. This was done by evaluating the annuli and the central cores of the constructs separately. METHOD: Chondrocyte-seeded agarose hydrogels were grown in free-swelling and dynamic, unconfined loading cultures for 42 days. After mechanical testing, constructs were allowed to recover for 1-2h, the central 3mm cores removed, and the cores and annuli were retested separately. Histological and/or biochemical analyses for DNA, glycosaminoglycan (GAG), collagen, type I collagen, type II collagen, and elastin were performed. Multiple regression analysis was used to determine the correlation between the biochemical and material properties of the constructs. RESULTS: The cores and annuli of chondrocyte-seeded constructs did not exhibit significant differences in material properties and GAG content. Annuli possessed greater DNA and collagen content over time in culture than cores. Dynamic loading enhanced the material properties and GAG content of cores, annuli, and whole constructs relative to free-swelling controls, but it did not alter the radial variations compared to free-swelling culture. CONCLUSION: Surprisingly, the benefits of dynamic loading on tissue properties extended through the entire construct and did not result in radial variations as measured via the coring technique in this study. Nutrient transport limitations and the formation of a fibrous capsule on the periphery may explain the differences in DNA and collagen between cores and annuli. No differences in GAG distribution may be due to sufficient chemical signals and building blocks for GAG synthesis throughout the constructs.

Influence of decreasing nutrient path length on the development of engineered cartilage.

OBJECTIVE: Chondrocyte-seeded agarose constructs of 4mm diameter (2.34 mm thickness) develop spatially inhomogeneous material properties with stiffer outer edges and a softer central core suggesting nutrient diffusion limitations to the central construct region [Guilak F, Sah RL, Setton LA. Physical regulation of cartilage metabolism. In: Mow VC, Hayes WC, Eds. Basic Orthopaedic Biomechanics, Philadelphia 1997;179-207.]. The effects of reducing construct thickness and creating channels running through the depth of the thick constructs were examined. METHODS: In Study 1, the properties of engineered cartilage of 0.78 mm (thin) or 2.34 mm (thick) thickness were compared. In Study 2, a single nutrient channel (1 mm diameter) was created in the middle of each thick construct. In Study 3, the effects of channels on larger 10 mm diameter, thick constructs were examined. RESULTS: Thin constructs developed superior mechanical and biochemical properties than thick constructs. The channeled constructs developed significantly higher mechanical properties vs control channel-free constructs while exhibiting similar glycosaminoglycan (GAG) and collagen content. Collagen staining suggested that channels resulted in a more uniform fibrillar network. Improvements in constructs of 10 mm diameter were similarly observed. CONCLUSIONS: This study demonstrated that more homogeneous tissue-engineered cartilage constructs with improved mechanical properties can be achieved by reducing their thickness or incorporating macroscopic nutrient channels. Our data further suggests that these macroscopic channels remain open long enough to promote this enhanced tissue development while exhibiting the potential to refill with cell elaborated matrix with additional culture time. Together with reports that <3 mm defects in cartilage heal in vivo and that irregular holes are associated with clinically used osteochondral graft procedures, we anticipate that a strategy of incorporating macroscopic channels may aid the development of clinically relevant engineered cartilage with functional properties.

Mechanical and biochemical characterization of cartilage explants in serum-free culture.

Allografts of articular cartilage are both used clinically for tissue-transplantation procedures and experimentally as model systems to study the physiological behavior of chondrocytes in their native extracellular matrix. Long-term maintenance of allograft tissue is challenging. Chemical mediators in poorly defined culture media can stimulate cells to quickly degrade their surrounding extracellular matrix. This is particularly true of juvenile cartilage which is generally more responsive to chemical stimuli than mature tissue. By carefully modulating the culture media, however, it may be possible to preserve allograft tissue over the long-term while maintaining its original mechanical and biochemical properties. In this study juvenile bovine cartilage explants (both chondral and osteochondral) were cultured in both chemically defined medium and serum-supplemented medium for up to 6 weeks. The mechanical properties and biochemical content of explants cultured in chemically defined medium were enhanced after 2 weeks in culture and thereafter remained stable with no loss of cell viability. In contrast, the mechanical properties of explants in serum-supplemented medium were degraded by ( approximately 70%) along with a concurrent loss of biochemical content (30-40% GAG). These results suggest that long-term maintenance of allografts can be extended significantly by the use of a chemically defined medium.

Effects of sustained interstitial fluid pressurization under migrating contact area, and boundary lubrication by synovial fluid, on cartilage friction.

OBJECTIVE: This experimental study tests two hypotheses which address outstanding questions in cartilage lubrication: can the friction coefficient remain low under sustained physiological loading conditions? How effective is synovial fluid (SF) in the lubrication of articular cartilage? Based on theory, it is hypothesized that migrating contact areas can maintain elevated cartilage interstitial fluid pressurization, thus a low friction coefficient, indefinitely. It is also hypothesized that the beneficial effects of SF stem from boundary lubrication rather than fluid-film lubrication. DESIGN: Five experiments were conducted on immature bovine femoro-tibial joints, to compare the frictional response under migrating vs stationary contact areas; the frictional response in SF vs saline; the role of sliding velocity and the role of congruence on the friction coefficient. RESULTS: Migrating contact area could maintain a low friction coefficient under sustained physiological conditions of loading for at least 1 h. SF reduced the friction coefficient by a factor of approximately 1.5 relative to saline. However, interstitial fluid pressurization was far more effective, reducing the friction coefficient by a factor of approximately 60 relative to equilibrium (zero-pressure) conditions. It was confirmed that SF acts as a boundary lubricant. CONCLUSIONS: These results emphasize the importance of interstitial fluid pressurization on the frictional response of cartilage. They imply that the mechanical integrity of cartilage must be maintained to produce low friction in articular joints. The more limited effectiveness of SF implies that intra-articular injections of lubricants in degenerated joints may have only limited effectiveness on their tribological properties.

Amino acids supply in culture media is not a limiting factor in the matrix synthesis of engineered cartilage tissue.

Increased amino acid supplementation (0.5 x, 1.0 x, and 5.0 x recommended concentrations or additional proline) was hypothesized to increase the collagen content in engineered cartilage. No significant differences were found between groups in matrix content or dynamic modulus. Control constructs possessed the highest compressive Young's modulus on day 42. On day 42, compared to controls, decreased type II collagen was found with 0.5 x, 1.0 x, and 5.0 x supplementation and significantly increased DNA content found in 1.0 x and 5.0 x. No effects were observed on these measures with added proline. These results lead us to reject our hypothesis and indicate that the low collagen synthesis in engineered cartilage is not due to a limited supply of amino acids in media but may require a further stimulatory signal. The results of this study also highlight the impact that culture environment can play on the development of engineered cartilage.

The beneficial effect of delayed compressive loading on tissue-engineered cartilage constructs cultured with TGF-beta3.

OBJECTIVE: To determine whether the functional properties of tissue-engineered constructs cultured in a chemically-defined medium supplemented briefly with TGF-beta3 can be enhanced with the application of dynamic deformational loading. METHODS: Primary immature bovine cells (2-3 months old) were encapsulated in agarose hydrogel (2%, 30 x 10(6)cells/ml) and cultured in chemically-defined medium supplemented for the first 2 weeks with transforming growth factor beta 3 (TGF-beta3) (10 microg/ml). Physiologic deformational loading (1 Hz, 3 h/day, 10% unconfined deformation initially and tapering to 2% peak-to-peak deformation by day 42) was applied either concurrent with or after the period of TGF-beta3 supplementation. Mechanical and biochemical properties were evaluated up to day 56. RESULTS: Dynamic deformational loading applied concurrently with TGF-beta3 supplementation yielded significantly lower (-90%) overall mechanical properties when compared to free-swelling controls. In contrast, the same loading protocol applied after the discontinuation of the growth factor resulted in significantly increased (+10%) overall mechanical properties relative to free-swelling controls. Equilibrium modulus values reach 1306+/-79 kPa and glycosaminoglycan levels reach 8.7+/-1.6% w.w. during this 8-week period and are similar to host cartilage properties (994+/-280 kPa, 6.3+/-0.9% w.w.). CONCLUSIONS: An optimal strategy for the functional tissue engineering of articular cartilage, particularly to accelerate construct development, may incorporate sequential application of different growth factors and applied deformational loading.

The effect of finite compressive strain on chondrocyte viability in statically loaded bovine articular cartilage.

Recent studies have reported that certain regimes of compressive loading of articular cartilage result in increased cell death in the superficial tangential zone (STZ). The objectives of this study were (1) to test the prevalent hypothesis that preferential cell death in the STZ results from excessive compressive strain in that zone, relative to the middle and deep zones, by determining whether cell death correlates with the magnitude of compressive strain and (2) to test the corollary hypothesis that the viability response of cells is uniform through the thickness of the articular layer when exposed to the same loading environment. Live cartilage explants were statically compressed by approximately 65% of their original thickness, either normal to the articular surface (axial loading) or parallel to it (transverse loading). Cell viability after 12 h was compared to the local strain distribution measured by digital image correlation. Results showed that the strain distribution in the axially loaded samples was highest in the STZ (77%) and lowest in the deep zone (55%), whereas the strain was uniformly distributed in the transversely loaded samples (64%). In contrast, axially and transversely loaded samples exhibited very similar profiles of cell death through the depth, with a preferential distribution in the STZ. Unloaded control samples showed negligible cell death. Thus, under prolonged static loading, depth-dependent variations in chondrocyte death did not correlate with the local depth-dependent compressive strain, and the prevalent hypothesis must be rejected. An alternative hypothesis, suggested by these results, is that superficial zone chondrocytes are more vulnerable to prolonged static loading than chondrocytes in the middle and deep zones.

A theoretical analysis of water transport through chondrocytes.

Because of the avascular nature of adult cartilage, nutrients and waste products are transported to and from the chondrocytes by diffusion and convection through the extracellular matrix. The convective interstitial fluid flow within and around chondrocytes is poorly understood. This theoretical study demonstrates that the incorporation of a semi-permeable membrane when modeling the chondrocyte leads to the following findings: under mechanical loading of an isolated chondrocyte the intracellular fluid pressure is on the order of tens of Pascals and the transmembrane fluid outflow, on the order of picometers per second, takes several days to subside; consequently, the chondrocyte behaves practically as an incompressible solid whenever the loading duration is on the order of minutes or hours. When embedded in its extracellular matrix (ECM), the chondrocyte response is substantially different. Mechanical loading of the tissue leads to a fluid pressure difference between intracellular and extracellular compartments on the order of tens of kilopascals and the transmembrane outflow, on the order of a nanometer per second, subsides in about 1 h. The volume of the chondrocyte decreases concomitantly with that of the ECM. The interstitial fluid flow in the extracellular matrix is directed around the cell, with peak values on the order of tens of nanometers per second. The viscous fluid shear stress acting on the cell surface is several orders of magnitude smaller than the solid matrix shear stresses resulting from the ECM deformation. These results provide new insight toward our understanding of water transport in chondrocytes.

Removal of the superficial zone of bovine articular cartilage does not increase its frictional coefficient.

OBJECTIVE: To investigate the role of the superficial zone in regulating the frictional response of articular cartilage. This zone contains the superficial protein (SZP), a proteoglycan synthesized exclusively by superficial zone chondrocytes and implicated in reducing the friction coefficient of cartilage. DESIGN: Unconfined compression creep tests with sliding of cartilage against glass in saline were carried out on fresh bovine cylindrical plugs (slashed circle Ø6 mm, n=35) obtained from 16 bovine shoulder joints (ages 1-3 months). In the first two experiments, friction tests were carried out before and after removal of the superficial zone ( approximately 100 microm), in a control and treatment group, using two different applied load magnitudes (4.4 N and 22.2 N). In the third experiment, friction tests were conducted on intact surfaces and the corresponding microtomed deep zone of the same specimen. RESULTS: In all tests the friction coefficient exhibited a transient response, increasing from a minimum value (mu(min)) to a near-equilibrium final value (micro(eq)). No statistical change (P>0.5) was found in micro(min) before and after removal of the superficial zone in both experiments 1 and 2. However, micro(eq) was observed to decrease significantly (P<0.001) after removal of the surface zone. Results from the third experiment confirm that micro(eq) is even lower at the deep zone. Surface roughness measurements with atomic force microscopy (AFM) revealed an increase in surface roughness after microtoming. Immunohistochemical staining confirmed the presence of SZP in intact specimens and its removal in microtomed specimens. CONCLUSIONS: The topmost ( approximately 100 microm) superficial zone of articular cartilage does not have special properties which enhances its frictional response.