Mechanobiology of Cartilage Impact Via Real-Time Metabolic Imaging.

Cartilage loading is important in both structural and biological contexts, with overloading known to cause osteoarthritis (OA). Cellular metabolism, which can be evaluated through the relative measures of glycolysis and oxidative phosphorylation, is important in disease processes across tissues. Details of structural damage coupled with cellular metabolism in cartilage have not been evaluated. Therefore, the aim of this study was to characterize the time- and location-dependent metabolic response to traumatic impact loading in articular cartilage. Cartilage samples from porcine femoral condyles underwent a single traumatic injury that created cracks in most samples. Before and up to 30 min after loading, samples underwent optical metabolic imaging. Optical metabolic imaging measures the fluorescent intensity of byproducts of the two metabolic pathways, flavin adenine dinucleotide for oxidative phosphorylation and nicotinamide adenine dinucleotide ± phosphate for glycolysis, as well as the redox ratio between them. Images were taken at varied distances from the center of the impact. Shortly after impact, fluorescence intensity in both channels decreased, while redox ratio was unchanged. The most dramatic metabolic response was measured closest to the impact center, with suppressed fluorescence in both channels relative to baseline. Redox ratio varied nonlinearly as a function of distance from the impact. Finally, both lower and higher magnitude loading reduced flavin adenine dinucleotide fluorescence, whereas reduced nicotinamide adenine dinucleotide ± phosphate fluorescence was associated only with low strain loads and high contact pressure loads, respectively. In conclusion, this study performed novel analysis of metabolic activity following induction of cartilage damage and demonstrated time-, distance-, and load-dependent response to traumatic impact loading.

Multiscale Strain as a Predictor of Impact-Induced Fissuring in Articular Cartilage.

Mechanical damage is central to both initiation and progression of osteoarthritis (OA). However, specific causal links between mechanics and cartilage damage are incompletely understood, which results in an inability to predict failure. The lack of understanding is primarily due to the difficulty in simultaneously resolving the high rates and small length scales relevant to the problem and in correlating such measurements to the resulting fissures. This study leveraged microscopy and high-speed imaging to resolve mechanics on the previously unexamined time and length scales of interest in cartilage damage, and used those mechanics to develop predictive models. The specific objectives of this study were to: first, quantify bulk and local mechanics during impact-induced fissuring; second, develop predictive models of fissuring based on bulk mechanics and local strain; and third, evaluate the accuracy of these models in predicting fissures. To achieve these three objectives, bovine tibial cartilage was impacted using a custom spring-loaded device mounted on an inverted microscope. The occurrence of fissures was modulated by varying impact energy. For the first objective, during impact, deformation was captured at 10,000 frames per second and bulk and local mechanics were analyzed. For the second objective, data from samples impacted with a 1.2 mm diameter rod were fit to logistic regression functions, creating models of fissure probability based on bulk and local mechanics. Finally, for the third objective, data from samples impacted with a 0.8 mm diameter rod were used to test the accuracy of model predictions. This study provides a direct comparison between bulk and local mechanical thresholds for the prediction of fissures in cartilage samples, and demonstrates that local mechanics provide more accurate predictions of local failure than bulk mechanics provide. Bulk mechanics were accurate predictors of fissure for the entire sample cohort, but poor predictors of fissure for individual samples. Local strain fields were highly heterogeneous and significant differences were determined between fissured and intact samples, indicating the presence of damage thresholds. In particular, first principal strain rate and maximum shear strain were the best predictors of local failure, as determined by concordance statistics. These data provide an important step in establishing causal links between local mechanics and cartilage damage; ultimately, data such as these can be used to link macro- and micro-scale mechanics and thereby predict mechanically mediated disease on a subject-specific basis.

Finite element prediction of transchondral stress and strain in the human hip.

Cartilage fissures, surface fibrillation, and delamination represent early signs of hip osteoarthritis (OA). This damage may be caused by elevated first principal (most tensile) strain and maximum shear stress. The objectives of this study were to use a population of validated finite element (FE) models of normal human hips to evaluate the required mesh for converged predictions of cartilage tensile strain and shear stress, to assess the sensitivity to cartilage constitutive assumptions, and to determine the patterns of transchondral stress and strain that occur during activities of daily living. Five specimen-specific FE models were evaluated using three constitutive models for articular cartilage: quasilinear neo-Hookean, nonlinear Veronda Westmann, and tension-compression nonlinear ellipsoidal fiber distribution (EFD). Transchondral predictions of maximum shear stress and first principal strain were determined. Mesh convergence analysis demonstrated that five trilinear elements were adequate through the depth of the cartilage for precise predictions. The EFD model had the stiffest response with increasing strains, predicting the largest peak stresses and smallest peak strains. Conversely, the neo-Hookean model predicted the smallest peak stresses and largest peak strains. Models with neo-Hookean cartilage predicted smaller transchondral gradients of maximum shear stress than those with Veronda Westmann and EFD models. For FE models with EFD cartilage, the anterolateral region of the acetabulum had larger peak maximum shear stress and first principal strain than all other anatomical regions, consistent with observations of cartilage damage in disease. Results demonstrate that tension-compression nonlinearity of a continuous fiber distribution exhibiting strain induced anisotropy incorporates important features that have large effects on predictions of transchondral stress and strain. This population of normal hips provides baseline data for future comparisons to pathomorphologic hips. This approach can be used to evaluate these and other mechanical variables in the human hip and their potential role in the pathogenesis of osteoarthritis (OA).

Failure in articular cartilage: Finite element predictions of stress, strain, and pressure under micro-indentation induced fracture.

Articular cartilage is found at the distal end of long bones and is responsible for assisting in joint articulation. While articular cartilage has remarkable resistance to failure, once initially damaged, degeneration is nearly irreversible. Thus, understanding damage initiation is important. There are a few proposed mechanisms for articular cartilage failure initiation: (A) a single collagen fibril stress-based regime; (B) a rate-dependent regime captured by brittle failure at slow displacement rates (SDR) and ductile failure at fast displacement rates (FDR); and (C) a rate-dependent regime where failure is governed by pressurization fragmentation at SDR and governed by strain at FDR. The objective of this study was to use finite element (FE) models to provide evidence to support or refute these proposed failure mechanisms. Models were developed of microfracture experiments that investigated osmolarity (hypo-osmolar, normal osmolarity, and hyper-osmolar) and displacement rate (FDR and SDR) effects. Cartilage was modeled with a neo-Hookean ground matrix, strain-dependent permeability, nonlinear fibril reinforcement with viscoelastic fibril terms, and Donnan equilibrium swelling. Total stress, solid matrix stress, Lagrange strain, and fluid pressure were determined under the indenter tip at the moment of microfracture. Results indicated significant rate dependence across multiple outputs, which does not support (A) a single failure regime. Larger solid and fluid pressures at FDR than SDR did not support (C) a rate-dependent regime split by pressurization at SDR and strain at FDR. Consistent solid shear stresses at SDR and consistent third principal solid stresses at FDR support (B) the ductile-brittle failure regime. These findings help to shed light on the underlying mechanisms of articular cartilage failure, which have implications for the development of osteoarthritis.

Subchondral bone fatigue injury in the parasagittal condylar grooves of the third metacarpal bone in thoroughbred racehorses elevates site-specific strain concentration.

Condylar stress fracture of the distal end of the third metacarpal/metatarsal (MC3/MT3) bones is a major cause of Thoroughbred racehorse injury and euthanasia worldwide. Functional adaptation to exercise and fatigue damage lead to structural changes in the subchondral bone that include increased modeling (resulting in sclerotic bone tissue) and targeted remodeling repair (resulting in focal resorption spaces in the parasagittal groove). Whether these focal structural changes, as detectable by standing computed tomography (sCT), lead to elevated strain at the common site of condylar stress fracture has not been demonstrated. Therefore, the goal of the present study was to compare full-field three-dimensional (3D) strain on the distopalmar aspect of MC3 bone specimens with and without focal subchondral bone injury (SBI). Thirteen forelimb specimens were collected from racing Thoroughbreds for mechanical testing ex vivo and underwent sCT. Subsequently, full-field displacement and strain at the joint surface were determined using stereo digital image correlation. Strain concentration was observed in the parasagittal groove (PSG) of the loaded condyles, and those with SBI in the PSG showed higher strain rates in this region than control bones. PSG strain rate in condyles with PSG SBI was more sensitive to CT density distribution in comparison with condyles with no sCT-detectable injury. Findings from this study help to interpret structural changes in the subchondral bone due to fatigue damage and to assess risk of incipient stress fracture in a patient-specific manner.

Comparison of radiography and computed tomography for identification of third metacarpal structural change and associated assessment of condylar stress fracture risk in Thoroughbred racehorses.

BACKGROUND: Catastrophic injury has a low incidence but leads to the death of many Thoroughbred racehorses. OBJECTIVES: To determine sensitivity, specificity, and reliability for third metacarpal condylar stress fracture risk assessment from digital radiographs (DR) and standing computed tomography (sCT). STUDY DESIGN: Controlled ex vivo experiment. METHODS: A blinded set of metacarpophalangeal joint DR and sCT images were prepared from 31 Thoroughbreds. Four observers evaluated the condyles and parasagittal grooves (PSG) of the third metacarpal bone for the extent of dense bone and lucency/fissure and assigned a risk assessment grade for condylar stress fracture based on imaging features. Sensitivity and specificity for detection of subchondral structural changes in the condyles and PSG, and for risk assessment for condylar stress fracture were determined by comparison with a reference assessment based on sCT and joint surface examination. Agreement between observers and the reference assessment and reliability between observers were determined. Intra-observer repeatability was also assessed. RESULTS: Sensitivity for detection of structural change was lower than specificity for both imaging methods and all observers. For agreement with the reference assessment of structural change, correlation coefficients were generally below 0.5 for DR and 0.49-0.82 for sCT. For horses categorised as normal risk on reference assessment, observer assessment often agreed with the reference. Sensitivity for risk assessment was lower than specificity for all observers. For horses with a reference assessment of high risk of injury, observers generally underestimated risk. Diagnostic sensitivity of risk assessment was improved with sCT imaging, particularly for horses categorised as having elevated risk of injury from the reference assessment. Assessment repeatability and reliability was better with sCT than DR. MAIN LIMITATIONS: The ex vivo study design influenced DR image sets. CONCLUSIONS: Risk assessment through screening with diagnostic imaging is a promising approach to improve injury prevention in racing Thoroughbreds. Knowledge of sensitivity and specificity of fetlock lesion detection provides the critical guidance needed to improve racehorse screening programs. We found improved detection of MC3 subchondral structural change and risk assessment for condylar stress fracture with sCT ex vivo.

Synthesis and characterization of a matrix-metalloproteinase responsive silk-elastinlike protein polymer.

Silk-elastinlike protein polymers (SELPs) are recombinant polymers consisting of tandem repeats of silk (GAGAGS) and elastin (GVGVP) units. By modification of the length and composition of these repeats, the properties of SELP hydrogels can be controlled for specific applications including nucleic acid and virus delivery and tissue engineering. Here, the structure of SELPs is further modified to include a sequence that is sensitive to matrix-metalloproteinases (MMPs). MMPs are a ubiquitous family of extracellular matrix-modifying enzymes that are commonly associated with numerous vital processes. Increased levels of MMPs are found at high levels locally in many types of solid tumors. By modifying the SELP backbone with MMP-sensitive peptide sequences, a hydrogel that is degradable by MMPs was produced. The MMP-sensitivity of the polymer was examined by incubation with MMP-2 and MMP-9, which yielded complete cleavage of all full-length polymers by 36 hours and 48 hours, respectively, with no observable effect on unmodified SELP. Hydrogel sensitivity was tested by exposure to MMP-2 or MMP-9 for 2 weeks, during which samples were taken to analyze protein loss from the hydrogel and release of 100 nm fluorescent beads. Following the incubation period, hydrogels were tested in mechanical compression to examine the loss of hydrogel stiffness due to degradation. It was found that MMP-2 and MMP-9 caused 63% and 44% increased protein loss and 65% and 95% increased release from MMP-sensitive hydrogels, while the compressive modulus decreased by 41% and 29%. These results suggest the potential of MMP-responsive SELPs for localized delivery of bioactive agents where MMPs are overexpressed.

Subject-specific analysis of joint contact mechanics: application to the study of osteoarthritis and surgical planning.

Advances in computational mechanics, constitutive modeling, and techniques for subject-specific modeling have opened the door to patient-specific simulation of the relationships between joint mechanics and osteoarthritis (OA), as well as patient-specific preoperative planning. This article reviews the application of computational biomechanics to the simulation of joint contact mechanics as relevant to the study of OA. This review begins with background regarding OA and the mechanical causes of OA in the context of simulations of joint mechanics. The broad range of technical considerations in creating validated subject-specific whole joint models is discussed. The types of computational models available for the study of joint mechanics are reviewed. The types of constitutive models that are available for articular cartilage are reviewed, with special attention to choosing an appropriate constitutive model for the application at hand. Issues related to model generation are discussed, including acquisition of model geometry from volumetric image data and specific considerations for acquisition of computed tomography and magnetic resonance imaging data. Approaches to model validation are reviewed. The areas of parametric analysis, factorial design, and probabilistic analysis are reviewed in the context of simulations of joint contact mechanics. Following the review of technical considerations, the article details insights that have been obtained from computational models of joint mechanics for normal joints; patient populations; the study of specific aspects of joint mechanics relevant to OA, such as congruency and instability; and preoperative planning. Finally, future directions for research and application are summarized.

Morphological quantification of the maxillary canine tooth in the domestic dog (Canis lupus familiaris).

Canine tooth shape is known to vary with diet and killing behavior in wild animals and the relationship between form and function is driven in part by selective pressure. However, comparative investigation of the domestic dog (Canis lupus familiaris) is of interest. How do they compare to their wild counterparts? This study sought to quantify and characterize the morphology of the canine tooth in the domestic dog, and to provide a preliminary investigation into the variance in canine tooth morphology across individual dogs of varying breeds. Three-dimensional (3D) models generated from micro-computed tomography (µ-CT) studies of 10 mature maxillary canine teeth from the domesticated dog (Canis lupus familiaris) were used to quantify key morphological features and evaluate variance among dogs. Results show that, utilizing modern imaging and model building software, the morphology of the canine tooth can be comprehensively characterized and quantified. Morphological variables such as second moment of area and section modulus (geometrical parameters related to resistance to bending), as well as aspect ratio, ridge sharpness, cusp sharpness and enamel thickness are optimized in biomechanically critical areas of the tooth crown to balance form and function. Tooth diameter, second moment of area, section modulus, cross sectional area, tooth volume and length as well as enamel thickness are highly correlated with body weight. In addition, we found preliminary evidence of morphological variance across individual dogs. Quantification of these features provide insight into the balance of form and function of the canine tooth in wild and domesticated canids. In addition, results suggest that variance between dogs exist in some morphological features and most morphological features are highly correlated with body weight.

Effect of mineral diets on the development of cartilage material properties.

Vitamin D and minerals, including zinc (Zn) and manganese (Mn), are vital in the development of bones, but their roles in the development of articular cartilage material behavior are not well understood. In this study, articular cartilage material properties from a hypovitaminosis D porcine model were evaluated. Pigs were produced by sows fed vitamin D deficient diets during gestation and lactation, and the offspring were subsequently fed vitamin D deficient diets for 3 weeks during the nursery period. Pigs were then assigned to dietary treatment groups with inorganic minerals only or inorganic plus organic (chelated) minerals. Humeral heads were harvested from pigs at 24 weeks of age. Linear elastic modulus and dissipated energy were measured under compression to 15% engineering strain at 1 Hz. Anatomical location within the humeral head affected elastic modulus. Diet significantly affected linear modulus and dissipated energy. The largest modulus and highest energy dissipation was in the inorganic zinc and manganese group; the lowest modulus and the least energy dissipation was in the organic (chelated) zinc and manganese group. Pairwise results between the control group and all vitamin D deficient groups were not statistically significant. Overall, these results suggest that mineral availability during rapid growth subsequent to a vitamin-D deficiency during gestation and lactation had minimal effects on articular cartilage material properties in young growing pigs. Though not statistically significant, some of the numerical differences between mineral sources suggest the potential importance of mineral availability during cartilage formation and warrant further study.

Correlation of mineral density and elastic modulus of dog dentin using µ-CT and nanoindentation.

This study sought to 1) investigate the spatial distribution of mineral density of dog dentin using µ-CT and 2) characterize the relationship between the elastic modulus and mineral density of dog dentin using nanoindentation and µ-CT. Maxillary canine teeth of 10 mature dogs were scanned with a µ-CT then sectioned in the transverse and vertical planes and tested using nanoindentation. Spatial distribution of mineral density and elastic modulus was quantified. Results demonstrated significant spatial variation in mineral density and elastic modulus. Mineral density and elastic modulus generally increased from the dentin-pulp interface to the dentino-enamel junction and from the crown base to the crown tip. Significant site dependent correlations between mineral density and elastic modulus were determined (0.021 > R2 > 0.408). The results of this study suggest that while mineral density is a mediator of elastic modulus, other mediators such as collagen content may contribute to the mechanical behavior of dog dentin.

Effect of osmolarity and displacement rate on cartilage microfracture clusters failure into two regimes.

Articular cartilage is a poroviscoelastic (PVE) material with remarkable resistance to fracture and fatigue failure. Cartilage failure mechanisms and material properties that govern failure are incompletely understood. Because cartilage is partially comprised of negatively charged glycosaminoglycans, altering solvent osmolarity can influence PVE relaxations. Therefore, this study aims to use osmolarity as a tool to provide additional data to interpret the role of PVE relaxations and identify cartilage failure regimes. Cartilage fracture was induced using a 100 µm radius spheroconical indenter at controlled displacement rates under three different osmolarity solvents. Secondarily, contact pressure (CP) and strain energy density (SED) were estimated to cluster data into two failure regimes with an expectation maximization algorithm. Critical displacement, critical load, critical time, and critical work to fracture increased with increasing osmolarity at a slow displacement rate whereas no significant effect was observed at a fast displacement rate. Clustering provided two distinct failure regimes, with regime (I) at lower normalized thickness (contact radius divided by sample thickness), and regime (II) at higher normalized thickness. Varied CP and SED in regime (I) suggest that failure in the regime is strain-governed. Constant CP and SED in regime (II) suggests that failure in the regime is dominantly governed by stress. These regimes can be interpreted as ductile versus brittle, or using a pressurized fragmentation interpretation. These findings demonstrated fundamental failure properties and postulate failure regimes for articular cartilage.

Vitamin D deficiency and anatomical region alters porcine growth plate properties.

Ossification of growth plate cartilage mediates longitudinal extension of long bones. Biomechanical and biochemical disruptions of growth plate function may lead to abnormal bone growth. In humans and animals, severe dietary vitamin D deficiency can lead to rickets which features growth plate widening, resulting in abnormalities in growth. However, effects of marginal vitamin D deficiencies on growth plates are not well understood. The purpose of this study was to examine the effects of a vitamin D deficient diet in the 26-day nursery phase on mechanical properties (ultimate normal stress, ultimate shear stress, ultimate strain, and tangent modulus) of porcine growth plate. Standard uniaxial tensile tests were applied on bone-growth plate-bone sections and the total stress was decomposed into normal stress and shear stress. Ultimate shear stress and ultimate strain traits were lower in the vitamin D deficient group than in the control. Regional differences were observed in all four variables. Ultimate normal stress was higher in the anterior region, which was consistent with a previous study. Sex differences were detected in ultimate normal stress, which was higher in females than in males. Interestingly, the classical finding of growth plate widening seen in severe vitamin D deficiency was not observed in the pigs with marginal vitamin D deficiency utilized in this study.

Maturation- and degeneration-dependent articular cartilage metabolism via optical redox ratio imaging.

From the two metabolic processes in healthy cartilage, glycolysis has been associated with proliferation and oxidative phosphorylation (oxphos) with matrix synthesis. Recently, metabolic dysregulation was significantly correlated with cartilage degradation and osteoarthritis progression. While these findings suggest maturation predisposes cartilage to metabolic instability with consequences for tissue maintenance, these links have not been shown. Therefore, this study sought to address three hypotheses (a) chondrocytes exhibit differential metabolic activity between immaturity (0-4 months), adolescence (5-18 months), and maturity (>18 months); (b) perturbation of metabolic activity has consequences on expression of genes pertinent to cartilage tissue maintenance; and (c) severity of cartilage damage is positively correlated with glycolysis and oxphos activity as well as optical redox ratio in postadolescent cartilage. Porcine femoral cartilage samples from pigs (3 days to 6 years) underwent optical redox ratio imaging, which measures autofluorescence of NAD(P)H and FAD. Gene expression analysis and histological scoring was conducted for comparison against imaging metrics. NAD(P)H and FAD autofluorescence both demonstrated increasing intensity with age, while optical redox ratio was lowest in adolescent samples compared to immature or mature samples. Inhibition of glycolysis suppressed expression of Col2, Col1, ADAMTS4, and ADAMTS5, while oxphos inhibition had no effect. FAD fluorescence and optical redox ratio were positively correlated with histological degeneration. This study demonstrates maturation- and degeneration-dependent metabolic activity in cartilage and explores the consequences of this differential activity on gene expression. This study aids our basic understanding of cartilage biology and highlights opportunity for potential diagnostic applications.

Effects of solvent osmolarity and viscosity on cartilage energy dissipation under high-frequency loading.

Articular cartilage is a spatially heterogeneous, dissipative biological hydrogel with a high fluid volume fraction. Although energy dissipation is important in the context of delaying cartilage damage, the dynamic behavior of articular cartilage equilibrated in media of varied osmolarity and viscosity is not widely understood. This study investigated the mechanical behaviors of cartilage when equilibrated to media of varying osmolarity and viscosity. Dynamic moduli and phase shift were measured at both low (1 Hz) and high (75-300 Hz) frequency, with cartilage samples compressed to varied offset strain levels. Increasing solution osmolarity and viscosity both independently resulted in larger energy dissipation and decreased dynamic modulus of cartilage at both low and high frequency. Mechanical property alterations induced by varying osmolarity are likely due to the change in permeability and fluid volume fraction within the tissue. The effects of solution viscosity are likely due to frictional interactions at the solid-fluid interface, affecting energy dissipation. These findings highlight the significance of interstitial fluid on the energy dissipation capabilities of the tissue, which can influence the onset of cartilage damage.

Structure-function relationships in dog dentin.

Investigations into teeth mechanical properties provide insight into physiological functions and pathological changes. This study sought to 1) quantify the spatial distribution of elastic modulus, hardness and the microstructural features of dog dentin and to 2) investigate quantitative relationships between the mechanical properties and the complex microstructure of dog dentin. Maxillary canine teeth of 10 mature dogs were sectioned in the transverse and vertical planes, then tested using nanoindentation and scanning electron microscopy (SEM). Microstructural features (dentin area fraction and dentinal tubule density) and mechanical properties (elastic modulus and hardness) were quantified. Results demonstrated significant anisotropy and spatial variation in elastic modulus, hardness, dentin area fraction and tubule density. These spatial variations adhered to a consistent distribution pattern; hardness, elastic modulus and dentin area fraction generally decreased from superficial to deep dentin and from crown tip to base; tubule density generally increased from superficial to deep dentin. Poor to moderate correlations between microstructural features and mechanical properties (R2 = 0.032-0.466) were determined. The results of this study suggest that the other constituents may contribute to the mechanical behavior of mammalian dentin. Our results also present several remaining opportunities for further investigation into the roles of organic components (e.g., collagen) and mineral content on dentin mechanical behavior.

Microindentation Technique to Create Localized Cartilage Microfractures.

Articular cartilage is a multiphasic, anisotropic, and heterogeneous material. Although cartilage possesses excellent mechanical and biological properties, it can undergo mechanical damage, resulting in osteoarthritis. Thus, it is important to understand the microscale failure behavior of cartilage in both basic science and clinical contexts. Determining cartilage failure behavior and mechanisms provides insight for improving treatment strategies to delay osteoarthritis initiation or progression and can also enhance the value of cartilage as bioinspiration for material fabrication. To investigate microscale failure behavior, we developed a protocol to initiate fractures by applying a microindentation technique using a well-defined tip geometry that creates localized cracks across a range of loading rates. The protocol includes extracting the tissue from the joint, preparing samples, and microfracture. Various aspects of the experiment, such as loading profile and solvent, can be adjusted to mimic physiological or pathological conditions and thereby further clarify phenomena underlying articular cartilage failure. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Harvesting and dissection of the joint surfaces Basic Protocol 2: Preparation of samples for microindentation and fatigue testing Basic Protocol 3: Microfracture using microindentation Basic Protocol 4: Crack propagation under cyclic loading.

Bi-component T2 mapping correlates with articular cartilage material properties.

Non-invasive estimation of cartilage material properties is useful for understanding cartilage health and creating subject-specific computational models. Bi-component T2 mapping measured using Multi-Component Driven Equilibrium Single Shot Observation of T1 and T2 (mcDESPOT) is sensitive for detecting cartilage degeneration within the human knee joint, but has not been correlated with cartilage composition and mechanical properties. Therefore, the purpose of this study was to investigate the relationship between bi-component T2 parameters measured using mcDESPOT at 3.0 T and cartilage composition and mechanical properties. Ex-vivo patellar cartilage specimens harvested from five human cadaveric knees were imaged using mcDESPOT at 3.0 T. Cartilage samples were removed from the patellae, mechanically tested to determine linear modulus and dissipated energy, and chemically tested to determine proteoglycan and collagen content. Parameter maps of single-component T2 relaxation time (T2), the T2 relaxation times of the fast relaxing macromolecular bound water component (T2F) and slow relaxing bulk water component (T2S), and the fraction of the fast relaxing macromolecular bound water component (FF) were compared to mechanical and chemical measures using linear regression. FF was significantly (p < 0.05) correlated with energy dissipation and linear modulus. T2 was significantly (p ≤ 0.05) correlated with elastic modulus at 1 Hz and energy dissipated at all frequencies. There were no other significant (p = 0.13-0.97) correlations between mcDESPOT parameters and mechanical properties. FF was significantly (p = 0.04) correlated with proteoglycan content. There were no other significant (p = 0.19-0.92) correlations between mcDESPOT parameters and proteoglycan or collagen content. This study suggests that FF measured using mcDESPOT at 3.0 T could be used to non-invasively estimate cartilage proteoglycan content, elastic modulus, and energy dissipation.

Rate-dependent adhesion of cartilage and its relation to relaxation mechanisms.

Cartilage adhesion has been found to play an important role in friction responses in the boundary lubrication regime, but its underlying mechanisms have only been partially understood. This study investigates the rate dependence of adhesion from pre-to post-relaxation timescales of cartilage and its possible relation to relaxation responses of the tissue. Adhesion tests on cartilage were performed to obtain rate-dependent cartilage adhesion from relaxed to unrelaxed states and corresponding relaxation responses. The rate dependence of cartilage adhesion was analyzed based on experimental relaxation responses. Cartilage adhesion increased about 20 times from relaxed to unrelaxed states. This rate-dependent enhancement correlated well with the load relaxation responses in a characteristic time domain. These experimental results indicated that the degree of recovery (or relaxation) in the vicinity of contact during unloading governed the rate dependence of cartilage adhesion. In addition, the experimentally measured enhancement of adhesion was interpreted with the aid of computationally and analytically predicted adhesion trends in viscoelastic, poroviscoelastic, and cohesive contact models. Agreement between the experimental and predicted trends implied that the enhancement of cartilage adhesion originated from complex combinations of interfacial peeling and negative fluid pressure generated within the contact area during unloading. These findings enhance the current understanding of rate-dependent adhesion mechanisms explored within short time scales and thus could provide new insight into friction responses and stick-induced damage in cartilage.

Interfibrillar shear behavior is altered in aging tendon fascicles.

Tendon elongation involves both stretching and sliding between adjacent fascicles and fibers. Hence, age-related changes in tendon matrix properties may alter sliding behavior and thereby affect injury thresholds. The objective of this study was to investigate the effects of age on interfibrillar shear behavior in partial cut tendon fascicles. Cine microscopic imaging was used to track deformation patterns of intact and partial cut fascicles from mature (9 months, n = 10) and aged (32 months, n = 10) rat tail tendons. Finite element (FE) models coupled with experimental data provided insight into age-related changes in tissue constitutive properties that could give rise to age-dependent behavior. Intact fascicles from aged tendons exhibited a 28% lower linear region modulus and reduced toe region when compared to fascicles from mature tendons. Partial cut tendon fascicles consistently exhibited a shearing plane that extended longitudinally from the tip of the cut. Both mature and aged fascicles exhibited distinct failure that was observable in differential displacement across the shearing plane. However, aged fascicles exhibited 11-20% higher grip-to-grip strain at failure and tended to exhibit more variable and greater differential displacement at failure, when compared to mature fascicles. FE models suggest that this age-related change in shear behavior arises from a reduction in interfibrillar shear modulus with age. These data suggest that aging alters interfibrillar failure mechanisms and hence may contribute to the increased propensity for injury that is commonly seen in older tendons.