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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

Glycosaminoglycan depletion increases energy dissipation in articular cartilage under high-frequency loading.

High-frequency material behavior of cartilage at macroscopic lengths is not widely understood, despite a wide range of frequencies and contact lengths experienced in vivo. For example, cartilage at different stages of matrix integrity can experience high-frequency loading during traumatic impact, making high-frequency behavior relevant in the context of structural failure. Therefore, this study examined macroscopic dissipative and mechanical responses of intact and glycosaminoglycan (GAG)-depleted cartilage under previously unexplored high-frequency loading. These dynamic responses were complemented with the evaluation of quasi-static responses. A custom dynamic mechanical analyzer was used to obtain dynamic behavior, and stress relaxation testing was performed to obtain quasi-static behavior. Under high-frequency loading, cartilage energy dissipation increased with GAG depletion and decreased with strain; dynamic modulus exhibited opposite trends. Similarly, under quasi-static loading, equilibrium modulus and relaxation time of cartilage decreased with GAG depletion. The increased energy dissipation after GAG depletion under high-frequency loading was likely due to increased viscoelastic dissipation. These findings broaden our understanding of fundamental properties of cartilage as a function of solid matrix integrity in an unprecedented loading regime. They also provide a foundation for analyzing energy dissipation associated with cartilage failure induced by traumatic impact.

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.

Maturity-dependent cartilage cell plasticity and sensitivity to external perturbation.

OBJECTIVE: Articular cartilage undergoes biological and morphological changes throughout maturation. The prevalence of osteoarthritis in the aged population suggests that maturation predisposes cartilage to degradation and/or impaired regeneration, but this process is not fully understood. Therefore, the objective of this study was to characterize the cellular and genetic profile of cartilage, as well as biological plasticity in response to mechanical and culture time stimuli, as a function of animal maturity. METHODS/DESIGN: Porcine articular cartilage explants were harvested from stifle joints of immature (2-4 weeks), adolescent (5-6 months), and mature (1-5 years) animals. Half of all samples were subjected to a single compressive mechanical load. Loaded samples were paired with unloaded controls for downstream analyses. Expression of cartilage progenitor cell markers CD105, CD44, and CD29 were determined via flow cytometry. Expression of matrix synthesis genes Col1, Col2, Col10, ACAN, and SOX9 were determined via qPCR. Tissue morphology and matrix content were examined histologically. Post-loading assays were performed immediately and following 7 days in culture. RESULTS: CD105 and CD29 expression decreased with maturity, while CD44 expression was upregulated in cartilage from mature animals. Expression of matrix synthesis genes were generally upregulated in cartilage from mature animals, and adolescent animals showed the lowest expression of several matrix synthesizing genes. Culture time and mechanical loading analyses revealed greater plasticity to mechanical loading and culture time in cartilage from younger animals. Histology confirmed distinct structural and biochemical profiles across maturity. CONCLUSION: This study demonstrates differential, nonlinear expression of chondroprogenitor markers and matrix synthesis genes as a function of cartilage maturity, as well as loss of biological plasticity in aged tissue. These findings have likely implications for age-related loss of regeneration and osteoarthritis progression.

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.

Subject-specific geometry affects acetabular contact pressure during gait more than subject-specific loading patterns.

Finite element modeling (FEM) can predict hip cartilage contact mechanics. This study investigated how subject-specific boundary conditions and joint geometry affect acetabular cartilage contact mechanics using a multi-scale workflow. For two healthy subjects, musculoskeletal models calculated subject-specific hip kinematics and loading, which were used as boundary conditions for FEM. Cartilage contact mechanics were predicted using either generic or subject-specific FEM and boundary conditions. A subject-specific mesh resulted in a more lateral contact. Effects of subject-specific boundary conditions varied between both subjects. Results highlight the complex interplay between loading and kinematics and their effect on cartilage contact mechanics.

Understanding the Stiff-to-Compliant Transition of the Meniscal Attachments by Spatial Correlation of Composition, Structure, and Mechanics.

Recently, the scientific community has shown considerable interest in engineering tissues with organized compositional and structural gradients to mimic hard-to-soft tissue interfaces. This effort is hindered by an incomplete understanding of the construction of native tissue interfaces. In this work, we combined Raman microscopy and confocal elastography to map compositional, structural, and mechanical features across the stiff-to-compliant interface of the attachments of the meniscus in the knee. This study provides new insight into the methods by which biology mediates multiple orders of magnitude changes in stiffness over tens of microns. We identified how the nano- to mesoscale architecture mediates complex microscale transitional regions across the interface: two regions defined by chemical composition, five distinguished by structural features, and three mechanically distinct regions. We identified three major components that lead to a robust interface between a soft tissue and bone: mobile collagen fiber units, a continuous interfacial region, and a local stiffness gradient. This tissue architecture allows for large displacements of collagen fibers in the attachments, enabling meniscal movement without localizing strains to the soft tissue-to-bone interface. The interplay of these regions reveals a method relying on hierarchical structuring across multiple length scales to minimize stress concentrators between highly dissimilar materials. These insights inspire new design strategies for synthetic soft tissue-to-bone attachments and biomimetic material interfaces.

Cartilage-on-cartilage cyclic loading induces mechanical and structural damage.

Cartilage breaks down during mechanically-mediated osteoarthritis (OA). While previous research has begun to elucidate mechanical, structural and cellular damage in response to cyclic loading, gaps remain in our understanding of the link between cyclic cartilage loading and OA-like mechanical damage. Thus, the aim of this study was to quantify irreversible cartilage damage in response to cyclic loading. A novel in vitro model of damage through cartilage-on-cartilage cyclic loading was established. Cartilage was loaded at 1 Hz to two different doses (10,000 or 50,000 cycles) between -6.0 ±â€¯0.2 MPa and -10.3 ±â€¯0.2 MPa 1st Piola-Kirchhoff stress. After loading, mechanical damage (altered mechanical properties: elastic moduli and dissipated energy) and structural damage (surface damage and specimen thickness) were quantified. Linear and tangential moduli were determined by fitting the loading portion of the stress-strain curves. Dissipated energy was calculated from the area between loading and unloading stress-strain curves. Specimen thickness was measured both before and after loading. Surface damage was assessed by staining samples with India ink, then imaging the articular surface. Cyclic loading resulted in dose-dependent decreases in linear and tangential moduli, energy dissipation, thickness, and intact area. Collectively, these results show that cartilage damage can be initiated by mechanical loading alone in vitro, suggesting that cyclic loading can cause in vivo damage. This study demonstrated that with increased number of cycles, cartilage undergoes both tissue softening and structural damage. These findings are a first step towards characterizing the cartilage response to cyclic loading, which can ultimately provide important insight for delaying the initiation and slowing the progression of OA.