Enzymatic digestion does not compromise sliding-mediated cartilage lubrication.

Articular cartilage's remarkable low-friction properties are essential to joint function. In osteoarthritis (OA), cartilage degeneration (e.g., proteoglycan loss and collagen damage) decreases tissue modulus and increases permeability. Although these changes impair lubrication in fully depressurized and slowly slid cartilage, new evidence suggests such relationships may not hold under biofidelic sliding conditions more representative of those encountered in vivo. Our recent studies using the convergent stationary contact area (cSCA) configuration demonstrate that articulation (i.e., sliding) generates interfacial hydrodynamic pressures capable of replenishing cartilage interstitial fluid/pressure lost to compressive loading through a mechanism termed tribological rehydration. This fluid recovery sustains in vivo-like kinetic friction coefficients (µk<0.02 in PBS and <0.005 in synovial fluid) with little sensitivity to mechanical properties in healthy tissue. However, the tribomechanical function of compromised cartilage under biofidelic sliding conditions remains unknown. Here, we investigated the effects of OA-like changes in cartilage mechanical properties, modeled via enzymatic digestion of mature bovine cartilage, on its tribomechanical function during cSCA sliding. We found no differences in sliding-driven tribological rehydration behaviors or µk between naïve and digested cSCA cartilage (in PBS or synovial fluid). This suggests that OA-like cartilage retains sufficient functional properties to support naïve-like fluid recovery and lubrication under biofidelic sliding conditions. However, OA-like cartilage accumulated greater total tissue strains due to elevated strain accrual during initial load application. Together, these results suggest that elevated total tissue strains-as opposed to activity-mediated strains or friction-driven wear-might be the key biomechanical mediator of OA pathology in cartilage. STATEMENT OF SIGNIFICANCE: Osteoarthritis (OA) decreases cartilage's modulus and increases its permeability. While these changes compromise frictional performance in benchtop testing under low fluid load support (FLS) conditions, whether such observations hold under sliding conditions that better represent the joints' dynamic FLS conditions in vivo is unclear. Here, we leveraged biofidelic benchtop sliding experiments-that is, those mimicking joints' native sliding environment-to examine how OA-like changes in mechanical properties effect cartilage's natural lubrication. We found no differences in sliding-mediated fluid recovery or kinetic friction behaviors between naïve and OA-like cartilage. However, OA-like cartilage experienced greater strain accumulation during load application, suggesting that elevated tissue strains (not friction-driven wear) may be the primary biomechanical mediator of OA pathology.

Exogenous Collagen Crosslinking is Highly Detrimental to Articular Cartilage Lubrication.

Healthy articular cartilage is a remarkable bearing material optimized for near-frictionless joint articulation. Because its limited self-repair capacity renders it susceptible to osteoarthritis (OA), approaches to reinforce or rebuild degenerative cartilage are of significant interest. While exogenous collagen crosslinking (CXL) treatments improve cartilage's mechanical properties and increase its resistance to enzymatic degradation, their effects on cartilage lubrication remain less clear. Here, we examined how the collagen crosslinking agents genipin (GP) and glutaraldehyde (GTA) impact cartilage lubrication using the convergent stationary contact area (cSCA) configuration. Unlike classical configurations, the cSCA sustains biofidelic kinetic friction coefficients (µk) via superposition of interstitial and hydrodynamic pressurization (i.e., tribological rehydration). As expected, glutaraldehyde- and genipin-mediated CXL increased cartilage's tensile and compressive moduli. Although net tribological rehydration was retained after CXL, GP or GTA treatment drastically elevated µk. Both healthy and "OA-like" cartilage (generated via enzymatic digestion) sustained remarkably low µk in saline- (≤0.02) and synovial fluid-lubricated contacts (≤0.006). After CXL, µk increased up to 30-fold, reaching values associated with marked chondrocyte death in vitro. These results demonstrate that mechanical properties (i.e., stiffness) are necessary, but not sufficient, metrics of cartilage function. Furthermore, the marked impairment in lubrication suggests that CXL-mediated stiffening is ill-suited to cartilage preservation or joint resurfacing.

Comparative tribology II-Measurable biphasic tissue properties have predictable impacts on cartilage rehydration and lubricity.

Healthy articular cartilage supports load bearing and frictional properties unmatched among biological tissues and man-made bearing materials. Balancing fluid exudation and recovery under loaded and articulated conditions is essential to the tissue's biological and mechanical longevity. Our prior tribological investigations, which leveraged the convergent stationary contact area (cSCA) configuration, revealed that sliding alone can modulate cartilage interstitial fluid pressurization and the recovery and maintenance of lubrication under load through a mechanism termed 'tribological rehydration.' Our recent comparative assessment of tribological rehydration revealed remarkably consistent sliding speed-dependent fluid recovery and lubrication behaviors across femoral condyle cartilage from five mammalian species (equine/horse, bovine/cow, porcine/pig, ovine/sheep, and caprine/goat). In the present study, we identified and characterized key predictive relationships among tissue properties, sliding-induced tribological rehydration, and the modulation/recovery of lubrication within healthy articular cartilage. Using correlational analysis, we linked observed speed-dependent tribological rehydration behaviors to cartilage's geometry and biphasic properties (tensile and compressive moduli, and permeability). Together, these findings demonstrate that easily measurable biphasic tissue characteristics (e.g., bulk tissue material properties, compressive strain magnitude, and strain rates) can be used to predict cartilage's rehydration and lubricating abilities, and ultimately its function in vivo. STATEMENT OF SIGNIFICANCE: In healthy cartilage, articulation recovers fluid lost to static loading thereby sustaining tissue lubricity. Osteoarthritis causes changes to cartilage composition, stiffness, and permeability associated with faster fluid exudation and presumably poorer frictional outcomes. Yet, the relationship between mechanical properties and fluid recovery during articulation/sliding remains unclear. Through innovative, high-speed benchtop sliding and indentation experiments, we found that cartilage's tissue properties regulate its exudation/hydration under slow sliding speeds but have minimal effect at high sliding speeds. In fact, cartilage rehydration appears insensitive to permeability and stiffness under high fluid load support conditions. This new understanding of the balance of cartilage exudation and rehydration during activity, based upon comparative tribology studies, may improve prevention and rehabilitation strategies for joint injuries and osteoarthritis.

The modes and competing rates of cartilage fluid loss and recovery.

Cartilage loses, recovers, and maintains its thickness, hydration, and biomechanical functions based on competing rates of fluid loss and recovery under varying joint-use conditions. While the mechanics and implications of load-induced fluid loss have been studied extensively, those of fluid recovery have not. This study isolates, quantifies, and compares rates of cartilage recovery from three known modes: (1) passive swelling - fluid recovery within a static unloaded contact area; (2) free swelling - unrestricted fluid recovery by an exposed surface; (3) tribological rehydration - fluid recovery within a loaded contact area during sliding. Following static loading of adult bovine articular cartilage to between 100 and 500 µm of compression, passive swelling, free swelling, and tribological rehydration exhibited average rates of 0.11 ± 0.04, 0.71 ± 0.15, and 0.63 ± 0.22 µm/s, respectively, over the first 100 s of recovery; for comparison, the mean exudation rate just prior to sliding was 0.06 ± 0.04 µm/s. For this range of compressions, we detected no significant difference between free swelling and tribological rehydration rates. However, free swelling and tribological rehydration rates, those associated with joint articulation, were ∼7-fold faster than passive swelling rates. While previous studies show how joint articulation prevents fluid loss indefinitely, this study shows that joint articulation reverses fluid loss following static loading at >10-fold the preceding exudation rate. These competitive recovery rates suggest that joint space and function may be best maintained throughout an otherwise sedentary day using brief but regular physical activity. STATEMENT OF SIGNIFICANCE: Cartilage loses, recovers, and maintains its thickness, hydration, and biomechanical functions based on competing rates of fluid loss and recovery under varying joint-use conditions. While load-induced fluid loss is extremely well studied, this is the first to define the competing modes of fluid recovery and to quantify their rates. The results show that the fluid recovery modes associated with joint articulation are 10-fold faster than exudation during static loading and passive swelling during static unloading. The results suggest that joint space and function are best maintained throughout an otherwise sedentary day using brief but regular physical activities.

Articular Cartilage Friction, Strain, and Viability Under Physiological to Pathological Benchtop Sliding Conditions.

In vivo, articular cartilage is exceptionally resistant to wear, damage, and dysfunction. However, replicating cartilage's phenomenal in vivo tribomechanics (i.e., high fluid load support, low frictions and strains) and mechanobiology on the benchtop has been difficult, because classical testing approaches tend to minimize hydrodynamic contributors to tissue function. Our convergent stationary contact area (cSCA) configuration retains the ability for hydrodynamically-mediated processes to contribute to interstitial hydration recovery and tribomechanical function via 'tribological rehydration'. Using the cSCA, we investigated how in situ chondrocyte survival is impacted by the presence of tribological rehydration during the reciprocal sliding of a glass counterface against a compressively loaded equine cSCA cartilage explant. When tribological rehydration was compromised during testing, by slow-speed sliding, 'pathophysiological' tribomechanical environments and high surface cell death were observed. When tribological rehydration was preserved, by high-speed sliding, 'semi-physiological' sliding environments and suppressed cell death were realized. Inclusion of synovial fluid during testing fostered 'truly physiological' sliding outcomes consistent with the in vivo environment but had limited influence on cell death compared to high-speed sliding in PBS. Subsequently, path analysis identified friction as a primary driver of cell death, with strain an indirect driver, supporting the contention that articulation mediated rehydration can benefit both the biomechanical properties and biological homeostasis of cartilage. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12195-021-00671-2.

Comparative Tribology: Articulation-induced rehydration of cartilage across species.

Articular cartilage is a robust tissue that facilitates load distribution and wear-free articulation in diarthrodial joints. These biomechanical capabilities are fundamentally tied to tissue hydration, whereby high interstitial fluid pressures and fluid load support facilitate the maintenance of low tissue strains and frictions. Our recent ex vivo studies of cartilage sliding biomechanics using the convergent stationary contact area (cSCA) configuration, first introduced by Dowson and colleagues, unexpectedly demonstrated that sliding alone can promote recovery of interstitial pressure and lubrication lost to static compression through a mechanism termed 'tribological rehydration.' Although exclusively examined in bovine stifle cartilage to date, we hypothesized that tribological rehydration, i.e., the ability to recover/modulate tissue strains and lubrication through sliding, is a universal behavior of articular cartilage. This study aimed to establish if, and to what extent, sliding-induced tribological rehydration is conserved in articular cartilage across a number of preclinical animal species/models and diarthrodial joints. Using a comparative approach, we found that articular cartilage from equine, bovine, ovine, and caprine stifles, and porcine stifle, hip, and tarsal joints all exhibited remarkably consistent sliding speed-dependent compression/strain recovery and lubrication behaviors under matched contact stresses (0.25 MPa). All cartilage specimens tested supported robust, tribological rehydration during high-speed sliding (>30 mm/s), which as a result of competitive recovery of interstitial lubrication, promoted remarkable decreases in kinetic friction during continuous sliding. The conservation of tribological rehydration across mammalian quadruped articular cartilage suggests that sliding-induced recovery of interstitial hydration represents an important tissue adaptation and largely understudied contributor to the biomechanics of cartilage and joints.

Detrimental effects of long sedentary bouts on the biomechanical response of cartilage to sliding.

Purpose/Aim: Epidemiological evidence suggests, contrary to popular mythos, that increased exercise/joint activity does not place articular cartilage at increased risk of disease, but instead promotes joint health. One explanation for this might be activity-induced cartilage rehydration; where joint articulation drives restoration of tissue hydration, thickness, and dependent tribomechanical outcomes (e.g., load support, stiffness, and lubricity) lost to joint loading. However, there have been no studies investigating how patterning of intermittent articulation influences the hydration and biomechanical functions of cartilage.Materials and Methods: Here we leveraged the convergent stationary contact area (cSCA) testing configuration and its unique ability to drive tribological rehydration, to elucidate how intermittency of activity affects the biomechanical functions of bovine stifle cartilage under well-controlled sliding conditions that have been designed to model a typical "day" of human joint activity.Results: For a fixed volume of "daily" activity (30 min) and sedentary time (60 min), breaking up intermittent activity into longer and less-frequent bouts (corresponding to longer continuous sedentary periods) resulted in the exposure of articular cartilage to markedly greater strains, losses of interstitial pressure, and friction coefficients.Conclusions: These results demonstrated that the regularity of ex vivo activity regimens, specifically the duration of sedentary bouts, had a dominant effect on the biomechanical functions of articular cartilage. In more practical terms, the results suggest that brief but regular movement patterns (e.g., every hour) may be biomechanically preferred to long and infrequent movement patterns (e.g., a long walk after a sedentary day) when controlling for daily activity volume (e.g., 30 min).