Contribution of collagen degradation and proteoglycan depletion to cartilage degeneration in primary and secondary osteoarthritis: an in silico study.

OBJECTIVES: Current experimental approaches cannot elucidate the effect of maladaptive changes on the main cartilage constituents during the degeneration process in osteoarthritis (OA). In silico approaches, however, allow creating 'virtual knock-out' cases to elucidate these effects in a constituent-specific manner. We used such an approach to study the main mechanisms of cartilage degeneration in different mechanical loadings associated with the following OA etiologies: (1) physiological loading of degenerated cartilage, (2) injurious loading of healthy intact cartilage and (3) physiological loading of cartilage with a focal defect. METHODS: We used the recently developed Cartilage Adaptive REorientation Degeneration (CARED) framework to simulate cartilage degeneration associated with primary and secondary OA (OA cases (1)-(3)). CARED incorporates numerical description of tissue-level cartilage degeneration mechanisms in OA, namely, collagen degradation, collagen reorientation, fixed charged density loss and tissue hydration increase following mechanical loading. We created 'virtual knock-out' scenarios by deactivating these degenerative processes one at a time in each of the three OA cases. RESULTS: In the injurious loading of intact and physiological loading of degenerated cartilage, collagen degradation drives degenerative changes through fixed charge density loss and tissue hydration rise. In contrast, the two later mechanisms were more prominent in the focal defect cartilage model. CONCLUSION: The virtual knock-out models reveal that injurious loading to intact cartilage and physiological loading to degenerated cartilage induce initial degenerative changes in the collagen network, whereas, in the presence of a focal cartilage defect, mechanical loading initially causes proteoglycans (PG) depletion, before changes in the collagen fibril network occur.

Machine learning augmented near-infrared spectroscopy: In vivo follow-up of cartilage defects.

OBJECTIVE: To assess the potential of near-infrared spectroscopy (NIRS) for in vivo arthroscopic monitoring of cartilage defects. METHOD: Sharp and blunt cartilage grooves were induced in the radiocarpal and intercarpal joints of Shetland ponies and monitored at baseline (0 weeks) and at three follow-up timepoints (11, 23, and 39 weeks) by measuring near-infrared spectra in vivo at and around the grooves. The animals were sacrificed after 39 weeks and the joints were harvested. Spectra were reacquired ex vivo to ensure reliability of in vivo measurements and for reference analyses. Additionally, cartilage thickness and instantaneous modulus were determined via computed tomography and mechanical testing, respectively. The relationship between the ex vivo spectra and cartilage reference properties was determined using convolutional neural network. RESULTS: In an independent test set, the trained networks yielded significant correlations for cartilage thickness (ρ = 0.473) and instantaneous modulus (ρ = 0.498). These networks were used to predict the reference properties at baseline and at follow-up time points. In the radiocarpal joint, cartilage thickness increased significantly with both groove types after baseline and remained swollen. Additionally, at 39 weeks, a significant difference was observed in cartilage thickness between controls and sharp grooves. For the instantaneous modulus, a significant decrease was observed with both groove types in the radiocarpal joint from baseline to 23 and 39 weeks. CONCLUSION: NIRS combined with machine learning enabled determination of cartilage properties in vivo, thereby providing longitudinal evaluation of post-intervention injury development. Additionally, radiocarpal joints were found more vulnerable to cartilage degeneration after damage than intercarpal joints.

Effect of centrifugal force on the development of articular neocartilage with bovine primary chondrocytes.

A lot has been invested into understanding how to assemble cartilage tissue in vitro and various designs have been developed to manufacture cartilage tissue with native-like biological properties. So far, no satisfactory design has been presented. Bovine primary chondrocytes are used to self-assemble scaffold-free constructs to investigate whether mechanical loading by centrifugal force would be useful in manufacturing cartilage tissue in vitro. Six million chondrocytes were laid on top of defatted bone disks placed inside an agarose well in 50-ml culture tubes. The constructs were centrifuged once or three times per day for 15 min at a centrifugal force of 771×g for up to 4 weeks. Control samples were cultured under the same conditions without exposure to centrifugation. The samples were analysed by (immuno)histochemistry, Fourier transform infrared imaging, micro-computed tomography, biochemical and gene expression analyses. Biomechanical testing was also performed. The centrifuged tissues had a more even surface covering a larger area of the bone disk. Fourier transform infrared imaging analysis indicated a higher concentration of collagen in the top and bottom edges in some of the centrifuged samples. Glycosaminoglycan contents increased along the culture, while collagen content remained at a rather constant level. Aggrecan and procollagen α1(II) gene expression levels had no significant differences, while procollagen α2(I) levels were increased significantly. Biomechanical analyses did not reveal remarkable changes. The centrifugation regimes lead to more uniform tissue constructs, whereas improved biological properties of the native tissue could not be obtained by centrifugation.

Functional effects of an interpenetrating polymer network on articular cartilage mechanical properties.

OBJECTIVE: Depletion of glycosaminoglycans (GAGs) and degradation of collagen network are early hallmarks of osteoarthritis (OA). Currently, there are no chondroprotective therapies that mitigate the loss of GAGs or effectively restore the collagen network. Recently, a novel polymeric cartilage supplement was described that forms a charged interpenetrating polymer network (IPN) reconstituting the hydrophilic properties of the extracellular matrix (ECM). To investigate the mechanism by which this hydrophilic IPN improves articular cartilage material properties, a finite element (FE) model is used to evaluate the IPN's effect on the fibrillar collagen network, nonfibrillar matrix, and interstitial fluid flow. METHODS: Bovine osteochondral plugs were degraded with chondroitinase ABC to selectively decrease GAG content. Samples were mechanically tested before and after IPN treatment using unconfined testing geometry and stress-relaxation protocol. Every measurement was modeled separately using a fibril-reinforced poroviscoelastic FE model. Measurement replication was achieved by optimizing the following model parameters: initial and strain-dependent fibril network modulus (Ef0, Efε, respectively), nonfibrillar matrix modulus (Enf), initial permeability (k0) and strain-dependent permeability factor (M). RESULTS: Based on the FE model results, treatment of native and GAG depleted cartilage with the hydrophilic IPN increases the ECM stiffness and impedes fluid flow. The IPN did not alter the stiffness of fibrillary network. Cartilage permeability and the strain-dependent permeability factor decreased with increasing IPN w/v%. CONCLUSIONS: The IPN reconstitutes cartilage material properties primarily by augmenting the hydrophilic ECM. This reinforcement of the solid phase also affects the fluid phase reestablishing low permeability.

Importance of Patella, Quadriceps Forces, and Depthwise Cartilage Structure on Knee Joint Motion and Cartilage Response During Gait.

In finite-element (FE) models of the knee joint, patella is often omitted. We investigated the importance of patella and quadriceps forces on the knee joint motion by creating an FE model of the subject's knee. In addition, depthwise strains and stresses in patellar cartilage with different tissue properties were determined. An FE model was created from subject's magnetic resonance images. Knee rotations, moments, and translational forces during gait were recorded in a motion laboratory and used as an input for the model. Three material models were implemented into the patellar cartilage: (1) homogeneous model, (2) inhomogeneous (arcadelike fibrils), and (3) random fibrils at the superficial zone, mimicking early stages of osteoarthritis (OA). Implementation of patella and quadriceps forces into the model substantially reduced the internal-external femoral rotations (versus without patella). The simulated rotations in the model with the patella matched the measured rotations at its best. In the inhomogeneous model, maximum principal stresses increased substantially in the middle zone of the cartilage. The early OA model showed increased compressive strains in the superficial and middle zones of the cartilage and decreased stresses and fibril strains especially in the middle zone. The results suggest that patella and quadriceps forces should be included in moment- and force-driven FE knee joint models. The results indicate that the middle zone has a major role in resisting shear forces in the patellar cartilage. Also, early degenerative changes in the collagen network substantially affect the cartilage depthwise response in the patella during walking.

Contrast enhanced imaging of human meniscus using cone beam CT.

OBJECTIVE: Meniscal injuries can lead to mechanical overloading of articular cartilage and eventually to knee osteoarthritis. The objective was to evaluate the potential of contrast enhanced computed tomography (CECT) to image contrast agent (CA) diffusion in human menisci with a clinical cone beam CT scanner. DESIGN: Isolated human menisci (n = 26) were imaged using magnetic resonance imaging (MRI) and CECT in situ. Diffusion of anionic CA into the meniscus was imaged for up to 30 h. The results of CECT were compared with water, collagen and proteoglycan (PG) contents, biomechanical properties, age and histological and MR images of the samples. RESULTS: Diffusion of CA required over 25 h to reach equilibrium. The CA partition (the CA concentration in the tissue divided by that in the bath) at the 40 min time point correlated significantly with that at the 30 h time point in both lateral (r = 0.706, P = 0.007) and medial (r = 0.669, P = 0.012) menisci. Furthermore, CA partition in meniscus after 30 h of diffusion agreed qualitatively with the distribution of PGs. CONCLUSION: The cross-sectional distribution of CA was consistent with that reported in a previous µCT study on bovine meniscus. The time required to reach diffusion equilibrium was found impractical for clinical applications. However, based on the present results, shorter delay between injection and imaging (e.g., 40 min) could be feasible in clinical diagnostics of meniscal pathologies.

Alterations in subchondral bone plate, trabecular bone and articular cartilage properties of rabbit femoral condyles at 4 weeks after anterior cruciate ligament transection.

OBJECTIVE: To quantify early osteoarthritic-like changes in the structure and volume of subchondral bone plate and trabecular bone and properties of articular cartilage in a rabbit model of osteoarthritis (OA) induced by anterior cruciate ligament transection (ACLT). METHODS: Left knee joints from eight skeletally mature New Zealand white rabbits underwent ACLT surgery, while the contralateral (CTRL) right knee joints were left unoperated. Femoral condyles were harvested 4 weeks after ACLT. Micro-computed tomography imaging was applied to evaluate the structural properties of subchondral bone plate and trabecular bone. Additionally, biomechanical properties, structure and composition of articular cartilage were assessed. RESULTS: As a result of ACLT, significant thinning of the subchondral bone plate (P < 0.05) was accompanied by significantly reduced trabecular bone volume fraction and trabecular thickness in the medial femoral condyle compartment (P < 0.05), while no changes were observed in the lateral compartment. In both lateral and medial femoral condyles, the equilibrium modulus and superficial zone proteoglycan (PG) content were significantly lower in ACLT than CTRL joint cartilage (P < 0.05). Significant alterations in the collagen orientation angle extended substantially deeper into cartilage from the ACLT joints in the lateral femoral condyle relative to the medial condyle compartment (P < 0.05). CONCLUSIONS: In this model of early OA, significant changes in volume and microstructure of subchondral bone plate and trabecular bone were detected only in the femoral medial condyle, while alterations in articular cartilage properties were more severe in the lateral compartment. The former finding may be associated with reduced joint loading in the medial compartment due to ACLT, while the latter finding reflects early osteoarthritic changes in the lateral compartment.

Estimation of articular cartilage properties using multivariate analysis of optical coherence tomography signal.

OBJECTIVE: The aim was to investigate the applicability of multivariate analysis of optical coherence tomography (OCT) information for determining structural integrity, composition and mechanical properties of articular cartilage. DESIGN: Equine osteochondral samples (N = 65) were imaged with OCT, and their total attenuation and backscattering coefficients (µt and µb) were measured. Subsequently, the Mankin score, optical density (OD) describing the fixed charge density, light absorbance in amide I region (Aamide), collagen orientation, permeability, fibril network modulus (Ef) and non-fibrillar matrix modulus (Em) of the samples were determined. Partial least squares (PLS) regression model was calculated to predict tissue properties from the OCT signals of the samples. RESULTS: Significant correlations between the measured and predicted mean collagen orientation (R(2) = 0.75, P < 0.0001), permeability (R(2) = 0.74, P < 0.0001), mean OD (R(2) = 0.73, P < 0.0001), Mankin scores (R(2) = 0.70, P < 0.0001), Em (R(2) = 0.50, P < 0.0001), Ef (R(2) = 0.42, P < 0.0001), and Aamide (R(2) = 0.43, P < 0.0001) were obtained. Significant correlation was also found between µb and Ef (ρ = 0.280, P = 0.03), but not between µt and any of the determined properties of articular cartilage (P > 0.05). CONCLUSION: Multivariate analysis of OCT signal provided good estimates for tissue structure, composition and mechanical properties. This technique may significantly enhance OCT evaluation of articular cartilage integrity, and could be applied, for example, in delineation of degenerated areas around cartilage injuries during arthroscopic repair surgery.

In vitro glycation of articular cartilage alters the biomechanical response of chondrocytes in a depth-dependent manner.

OBJECTIVE: To determine if increasing cartilage cross-links through in vitro glycation of cartilage explants can alter the biomechanical response of chondrocytes to compressive deformation. METHOD: Bovine osteochondral explants were either incubated with cell culture solution supplemented with (n = 7) or without (n = 7) ribose for 42 h in order to induce glycation. Deformation-induced changes in cell volume, dimensions and local tissue strains were determined through confocal laser scanning microscopy (CLSM) and the use of a custom built micro-compression device. Osteochondral explants were also utilized to demonstrate changes in depth-wise tissue properties, biomechanical tissue properties and cross-links such as pentosidine (Pent), hydroxylysyl pyridinoline (HP) and lysyl pyridinoline (LP). RESULTS: The ribose treated osteochondral samples experienced reduced cell volume deformation in the upper tissue zone by ∼ 8% (P = 0.005), as compared the control samples, through restricting cell expansion. In the deeper tissue zone, cell volume deformation was increased by ∼ 12% (P < 0.001) via the transmission of mechanical signals further into the tissue depth. Biomechanical testing of the ribose treated osteochondral samples demonstrated an increase in the equilibrium and dynamic strain dependent moduli (P < 0.001 and P = 0.008, respectively). The biochemical analysis revealed an increase in Pent cross-links (P < 0.001). Depth-wise tissue property analyses revealed increased levels of carbohydrate content, greater levels of fixed charge density and an increased carbohydrate to protein ratio from 6 to 16%, 55-100% and 72-79% of the normalized tissue thickness (from the surface), respectively, in the ribose-treated group (P < 0.05). CONCLUSION: In vitro glycation alters the biomechanical response of chondrocytes in cartilage differently in upper and deeper zones, offering possible insights into how aging could alter cell deformation behavior in cartilage.

Adiabatic rotating frame relaxation of MRI reveals early cartilage degeneration in a rabbit model of anterior cruciate ligament transection.

OBJECTIVE: To investigate the sensitivity of seven quantitative magnetic resonance imaging (MRI) parameters (adiabatic T1ρ, adiabatic T2ρ, continuous wave (CW) T1ρ, relaxation along a fictitious field (RAFF), T2 measured with adiabatic double echo (DE) and Carr-Purcell-Meiboom-Gill (CPMG) sequence, and T1 during off-resonance saturation [magnetization transfer (MT)]) to detect early osteoarthritic changes in a rabbit model of anterior cruciate ligament transection (ACLT). METHODS: ACLT was unilaterally induced in the knees of New Zealand White rabbits (n = 8) while contralateral joints served as controls. Femoral condyles of the joints were harvested 4 weeks post-ACLT. MRI was performed at 9.4 T. For reference, quantitative histology, Mankin grading and biomechanical measurements were conducted. RESULTS: Reference methods demonstrated early, superficial cartilage degeneration in the ACLT group, including significant loss of proteoglycans in both medial and lateral compartments, increased collagen fibril anisotropy in the lateral condyle and decreased biomechanical properties at both medial and lateral compartments. CW-T1ρ was prolonged in the lateral compartment of ACLT joints while adiabatic T1ρ and T2ρ detected degenerative changes in tissue in both lateral and medial condyles (P < 0.05). DE-T2 was significantly (P < 0.05) elevated only in the lateral compartment while CPMG-T2, MT or RAFF did not show a statistically significant difference between the groups. CONCLUSIONS: Adiabatic T1ρ and T2ρ relaxation times detected most sensitively early degenerative changes in cartilage 4 weeks post-ACLT in a rabbit model.

Site-dependent changes in structure and function of lapine articular cartilage 4 weeks after anterior cruciate ligament transection.

OBJECTIVE: The aim of this study was to investigate the site-dependent changes in the structure and function of articular cartilage in the lapine knee joint at a very early stage of osteoarthritis (OA), created experimentally by anterior cruciate ligament transection (ACLT). METHODS: Unilateral ACLT was performed in eight mature New Zealand white rabbits. ACL transected and contralateral (C-L) joints were prepared for analysis at 4 weeks after ACLT. Three rabbits with intact joints were used as a control group (CNTRL). Femoral groove, medial and lateral femoral condyles, and tibial plateaus were harvested and used in the analysis. Biomechanical tests, microscopy and spectroscopy were used to determine the biomechanical properties, composition and structure of the samples. A linear mixed model was chosen for statistical comparisons between the groups. RESULTS: As a result of ACLT, the equilibrium and dynamic moduli were decreased primarily in the femoral condyle cartilage. Up to three times lower moduli (P < 0.05) were observed in the ACLT group compared to the control group. Significant (P < 0.05) proteoglycan (PG) loss in the ACLT joint cartilage was observed up to a depth of 20-30% from the cartilage surface in femoral condyles, while significant PG loss was confined to more superficial regions in tibial plateaus and femoral groove. The collagen orientation angle was increased (P < 0.05) up to a cartilage depth of 60% by ACLT in the lateral femoral condyle, while smaller effects, but still significant, were observed at other locations. The collagen content was increased (P < 0.05) in the middle and deep zones of the ACLT group compared to the control group samples, especially in the lateral femoral condyle. CONCLUSION: Femoral condyle cartilage experienced the greatest structural and mechanical alterations in very early OA, as produced by ACLT. Degenerative alterations were observed especially in the superficial collagen fiber organization and PG content, while the collagen content was increased in the deep tissue of femoral condyle cartilage. The current findings provide novel information of the early stages of OA in different locations of the knee joint.

Experimental and numerical analysis of soft tissue stiffness measurement using manual indentation device--significance of indentation geometry and soft tissue thickness.

BACKGROUND: Indentation techniques haves been applied to measure stiffness of human soft tissues. Tissue properties and geometry of the indentation instrument control the measured response. METHODS: Mechanical roles of different soft tissues were characterized to understand the performance of the indentation instrument. An optimal instrument design was investigated. Experimental indentations in forearm of human subjects (N = 11) were conducted. Based on peripheral quantitative computed tomography imaging, a finite element (FE) model for indentation was created. The model response was matched with the experimental data. RESULTS: Optimized values for the elastic modulus of skin and adipose tissue were 130.2 and 2.5 kPa, respectively. The simulated indentation response was 3.9 ± 1.2 (mean ± SD) and 4.9 ± 2.0 times more sensitive to changes in the elastic modulus of the skin than to changes in the elastic modulus of adipose tissue and muscle, respectively. Skin thickness affected sensitivity of the instrument to detect changes in stiffness of the underlying tissues. CONCLUSION: Finite element modeling provides a feasible method to quantitatively evaluate the geometrical aspects and the sensitivity of an indentation measurement device. Systematically, the skin predominantly controlled the indentation response regardless of the indenter geometry or variations in the volume of different soft tissues.

Cell deformation behavior in mechanically loaded rabbit articular cartilage 4 weeks after anterior cruciate ligament transection.

OBJECTIVE: Chondrocyte stresses and strains in articular cartilage are known to modulate tissue mechanobiology. Cell deformation behavior in cartilage under mechanical loading is not known at the earliest stages of osteoarthritis. Thus, the aim of this study was to investigate the effect of mechanical loading on volume and morphology of chondrocytes in the superficial tissue of osteoarthritic cartilage obtained from anterior cruciate ligament transected (ACLT) rabbit knee joints, 4 weeks after intervention. METHODS: A unique custom-made microscopy indentation system with dual-photon microscope was used to apply controlled 2 MPa force-relaxation loading on patellar cartilage surfaces. Volume and morphology of chondrocytes were analyzed before and after loading. Also global and local tissue strains were calculated. Collagen content, collagen orientation and proteoglycan content were quantified with Fourier transform infrared microspectroscopy, polarized light microscopy and digital densitometry, respectively. RESULTS: Following the mechanical loading, the volume of chondrocytes in the superficial tissue increased significantly in ACLT cartilage by 24% (95% confidence interval (CI) 17.2-31.5, P < 0.001), while it reduced significantly in contralateral group tissue by -5.3% (95% CI -8.1 to -2.5, P = 0.003). Collagen content in ACLT and contralateral cartilage were similar. PG content was reduced and collagen orientation angle was increased in the superficial tissue of ACLT cartilage compared to the contralateral cartilage. CONCLUSIONS: We found the novel result that chondrocyte deformation behavior in the superficial tissue of rabbit articular cartilage is altered already at 4 weeks after ACLT, likely because of changes in collagen fibril orientation and a reduction in PG content.

Structure-function relationships in osteoarthritic human hip joint articular cartilage.

OBJECTIVES: It is currently poorly known how different structural and compositional components in human articular cartilage are related to their specific functional properties at different stages of osteoarthritis (OA). The objective of this study was to characterize the structure-function relationships of articular cartilage obtained from osteoarthritic human hip joints. METHODS: Articular cartilage samples with their subchondral bone (n = 15) were harvested during hip replacement surgeries from human femoral necks. Stress-relaxation tests, Mankin scoring, spectroscopic and microscopic methods were used to determine the biomechanical properties, OA grade, and the composition and structure of the samples. In order to obtain the mechanical material parameters for the samples, a fibril-reinforced poroviscoelastic model was fitted to the experimental data obtained from the stress-relaxation experiments. RESULTS: The strain-dependent collagen network modulus (E(f)(ε)) and the collagen orientation angle exhibited a negative linear correlation (r = -0.65, P < 0.01), while the permeability strain-dependency factor (M) and the collagen content exhibited a positive linear correlation (r = 0.56, P < 0.05). The nonfibrillar matrix modulus (E(nf)) also exhibited a positive linear correlation with the proteoglycan content (r = 0.54, P < 0.05). CONCLUSION: The study suggests that increased collagen orientation angle during OA primarily impairs the collagen network and the tensile stiffness of cartilage in a strain-dependent manner, while the decreased collagen content in OA facilitates fluid flow out of the tissue especially at high compressive strains. Thus, the results provide interesting and important information of the structure-function relationships of human hip joint cartilage and mechanisms during the progression of OA.

Computed tomography detects changes in contrast agent diffusion after collagen cross-linking typical to natural aging of articular cartilage.

OBJECTIVE: The effect of threose-induced collagen cross-linking on the mechanical and diffusive properties of cartilage was investigated in vitro. In particular, we investigated the potential of Contrast Enhanced Computed Tomography (CECT) to detect changes in articular cartilage after increased collagen cross-linking, which is an age-related phenomenon. METHODS: Osteochondral plugs (Ø=6.0 mm, n=28) were prepared from intact bovine patellae (n=7). Two of the four adjacent samples, prepared from each patella, were treated with threose to increase the collagen cross-linking, while the other two specimen served as paired controls. One sample pair was mechanically tested and then mechanically injured using a material testing device. Contrast agent [ioxaglate (Hexabrix™)] diffusion was imaged in the other specimen pair for 25 h using CECT. Water fraction, collagen and proteoglycan content, collagen network architecture and the amount of cross-links [hydroxylysyl pyridinoline (HP), lysyl pyridinoline (LP) and pentosidine (Pent)] of the samples were also determined. RESULTS: Cartilage collagen cross-linking, both Pent and LP, were significantly (P<0.001) increased due to threose treatment. CECT could detect the increased cross-links as the contrast agent penetration and the diffusion flux were significantly (P<0.05) lower in the threose treated than in untreated samples. The equilibrium modulus (+164%, P<0.05) and strain dependent dynamic modulus (+47%, P<0.05) were both significantly greater in the threose treated samples than in reference samples, but there was no association between the initial dynamic modulus and the threose treatment. The water fraction, proteoglycan and collagen contents, as well as collagen architecture, were not significantly altered by the threose treatment. CONCLUSIONS: To conclude, the CECT technique was found to be sensitive at detecting changes in cartilage tissue due to increased collagen cross-linking. This is important since increased cross-linking has been proposed to be related to the increased injury susceptibility of tissue.

Comparison of material models for anterior cruciate ligament in tension: from poroelastic to a novel fibril-reinforced nonlinear composite model.

Computational models of the knee joint are useful for evaluating stresses and strains within the joint tissues. However, the outcome of those models is sensitive to the material model and material properties chosen for ligaments, the collagen reinforced tissues connecting bone to bone. The purpose of this study was to investigate different compositionally motivated material models and further to develop a model that can accurately reproduce experimentally measured stress-relaxation data of bovine anterior cruciate ligament (ACL). Tensile testing samples were extracted from ACLs of bovine knee joints (N = 10) and subjected to a three-step stress-relaxation test at the toe region. Data from the experiments was averaged and one average finite element model was generated to replicate the experiment. Poroelastic and different fibril-reinforced poro(visco)elastic material models were applied, and their material parameters were optimized to reproduce the experimental force-time response. Material models with only fluid flow mediated relaxation were not able to capture the stress-relaxation behavior (R2 = 0.806, 0.803 and 0.938). The inclusion of the viscoelasticity of the fibrillar network improved the model prediction (R2 = 0.978 and 0.976), but the complex stress-relaxation behavior was best captured by a poroelastic model with a nonlinear two-relaxation-time strain-recruited viscoelastic fibrillar network (R2 = 0.997). The results suggest that in order to replicate the multi-step stress-relaxation behavior of ACL in tension, the fibrillar network formulation should include the complex nonlinear viscoelastic phenomena.

12 Degrees of Freedom Muscle Force Driven Fibril-Reinforced Poroviscoelastic Finite Element Model of the Knee Joint.

Accurate knowledge of the joint kinematics, kinetics, and soft tissue mechanical responses is essential in the evaluation of musculoskeletal (MS) disorders. Since in vivo measurement of these quantities requires invasive methods, musculoskeletal finite element (MSFE) models are widely used for simulations. There are, however, limitations in the current approaches. Sequentially linked MSFE models benefit from complex MS and FE models; however, MS model's outputs are independent of the FE model calculations. On the other hand, due to the computational burden, embedded (concurrent) MSFE models are limited to simple material models and cannot estimate detailed responses of the soft tissue. Thus, first we developed a MSFE model of the knee with a subject-specific MS model utilizing an embedded 12 degrees of freedom (DoFs) knee joint with elastic cartilages in which included both secondary kinematic and soft tissue deformations in the muscle force estimation (inverse dynamics). Then, a muscle-force-driven FE model with fibril-reinforced poroviscoelastic cartilages and fibril-reinforced poroelastic menisci was used in series to calculate detailed tissue mechanical responses (forward dynamics). Second, to demonstrate that our workflow improves the simulation results, outputs were compared to results from the same FE models which were driven by conventional MS models with a 1 DoF knee, with and without electromyography (EMG) assistance. The FE model driven by both the embedded and the EMG-assisted MS models estimated similar results and consistent with experiments from literature, compared to the results estimated by the FE model driven by the MS model with 1 DoF knee without EMG assistance.

Depth-wise progression of osteoarthritis in human articular cartilage: investigation of composition, structure and biomechanics.

OBJECTIVE: Osteoarthritis (OA) is characterized by the changes in structure and composition of articular cartilage. However, it is not fully known, what is the depth-wise change in two major components of the cartilage solid matrix, i.e., collagen and proteoglycans (PGs), during OA progression. Further, it is unknown how the depth-wise changes affect local tissue strains during compression. Our aim was to address these issues. METHODS: Data from the previous microscopic and biochemical measurements of the collagen content, distribution and orientation, PG content and distribution, water content and histological grade of normal and degenerated human patellar articular cartilage (n=73) were reanalyzed in a depth-wise manner. Using this information, a composition-based finite element (FE) model was used to estimate tissue function solely based on its composition and structure. RESULTS: The orientation angle of collagen fibrils in the superficial zone of cartilage was significantly less parallel to the surface (P<0.05) in samples with early degeneration than in healthy samples. Similarly, PG content was reduced in the superficial zone in early OA (P<0.05). However, collagen content decreased significantly only at the advanced stage of OA (P<0.05). The composition-based FE model showed that under a constant stress, local tissue strains increased as OA progressed. CONCLUSION: For the first time, depth-wise point-by-point statistical comparisons of structure and composition of human articular cartilage were conducted. The present results indicated that early OA is primarily characterized by the changes in collagen orientation and PG content in the superficial zone, while collagen content does not change until OA has progressed to its late stage. Our simulation results suggest that impact loads in OA joint could create a risk for tissue failure and cell death.

EMG-Assisted Muscle Force Driven Finite Element Model of the Knee Joint with Fibril-Reinforced Poroelastic Cartilages and Menisci.

Abnormal mechanical loading is essential in the onset and progression of knee osteoarthritis. Combined musculoskeletal (MS) and finite element (FE) modeling is a typical method to estimate load distribution and tissue responses in the knee joint. However, earlier combined models mostly utilize static-optimization based MS models and muscle force driven FE models typically use elastic materials for soft tissues or analyze specific time points of gait. Therefore, here we develop an electromyography-assisted muscle force driven FE model with fibril-reinforced poro(visco)elastic cartilages and menisci to analyze knee joint loading during the stance phase of gait. Moreover, since ligament pre-strains are one of the important uncertainties in joint modeling, we conducted a sensitivity analysis on the pre-strains of anterior and posterior cruciate ligaments (ACL and PCL) as well as medial and lateral collateral ligaments (MCL and LCL). The model produced kinematics and kinetics consistent with previous experimental data. Joint contact forces and contact areas were highly sensitive to ACL and PCL pre-strains, while those changed less cartilage stresses, fibril strains, and fluid pressures. The presented workflow could be used in a wide range of applications related to the aetiology of cartilage degeneration, optimization of rehabilitation exercises, and simulation of knee surgeries.

Comparison of water, hydroxyproline, uronic acid and elastin contents of bovine knee ligaments and patellar tendon and their relationships with biomechanical properties.

Mechanical material properties of ligaments originate from their biochemical composition and structural organization. However, it is not yet fully elucidated how biochemical contents vary between knee ligaments and patellar tendon (PT) and how they relate with mechanical properties. The purpose of this study was to compare water, collagen, proteoglycan and elastin contents between bovine knee ligaments and PT and correlate them with tensile material properties. Hydroxyproline (collagen), uronic acid (proteoglycan) and elastin contents per wet and dry weights were measured using colorimetric biochemical methods for bovine knee ligament and PT samples (n = 10 knees). Direct comparison and correlation with multiple linear regression were performed against biomechanical properties measured in our earlier study. Anterior cruciate ligament (ACL) and PT exhibited lower hydroxyproline content per wet weight compared with other ligaments (p < 0.05). Cruciate ligaments had higher uronic acid content per dry weight compared with collateral ligaments (p < 0.05). Posterior cruciate ligament had higher elastin content than ACL (p < 0.05). Higher hydroxyproline content per wet weight implied higher Young's modulus, strength and toughness. Quantitatively, higher elastin content per wet weight predicted higher toe region nonlinearity and Young's modulus whereas higher uronic acid content per dry weight predicted lower Young's modulus, yield stress and toughness. Differences between ligaments in biochemical composition highlight differences in their physiological function and loading regimes. As expected, collagen content showed similar trend with stiffness and strength. The predictive role of proteoglycan and elastin contents on the mechanical properties might indicate their important functional role in ligaments.