Cartilage-bone Plugs Research Articles

Effects of in vivo fatigue-induced microdamage on local subchondral bone strains

Biomechanical strain is a major stimulus of subchondral bone (SCB) tissue adaptation in joints but may also lead to initiation and propagation of microcracks, highlighting the importance of quantifying the intratissue strain in subchondral bone. In the present study, we used micro computed tomography (μCT) imaging, mechanical testing, and digital image correlation (DIC) techniques to evaluate the biomechanical strains in equine SCB under impact compression applied through the articular surface. We aimed to investigate the effects of in vivo accumulated microdamage in equine SCB on the distribution of mechanical impact strain through the articular cartilage. Under the applied strain of 2.0 ± 0.1% (mean ± standard deviation, n=15) to the articular surface of cartilage-bone plugs, the overall thickness of the SCB developed eSCBOverall = 0.7 ± 0.2% in all specimens. Contours of high strains in specimens without microdamage (NDmg) aligned parallel to the cartilage-bone interface with peak tensile, ϵt, and compressive, ϵc, strains of 0.5 ± 0.3% and 1.2 ± 0.4%, respectively at the time of peak compression (n=7). In damaged specimens (Dmg), contours of high strains aligned with the cracks in the imaged plane with peak strains of ϵt= 1.2 ± 0.8% and ϵc= 3.5 ± 2.2%, respectively (n=7). Microdamage was the main predictor of the normalised compressive and tensile strains across the SCB thickness. Results of multivariable analyses revealed presence of microdamage, distance from the articular surface and TMD were the main predictors of normalised compressive and tensile strain. Strain was greater in the superficial bone, particularly for specimens with microdamage. In vivo fatigue-induced microdamage is an important predictor of local subchondral bone strains.

Full-Field Strain Uncertainties and Residuals at the Cartilage-Bone Interface in Unstained Tissues Using Propagation-Based Phase-Contrast XCT and Digital Volume Correlation.

A deeper understanding of the cartilage-bone mechanics is fundamental to unravel onset and progression of osteoarthritis, enabling better diagnosis and treatment. The aim of this study is therefore to explore the capability of X-ray computed (XCT) phase-contrast imaging in a lab-based system to enable digital volume correlation (DVC) measurements of unstained cartilage-bone plugs from healthy adult bovines. DVC strain uncertainties were computed for both articular cartilage and mineralized tissue (calcified cartilage and subchondral bone) in the specimens at increasing propagation distances, ranging from absorption up to four times (4× such effective distance. In addition, a process of dehydration and rehydration was proposed to improve feature recognition in XCT of articular cartilage and mechanical properties of this tissue during the process were assessed via micromechanical probing (indentation), which was also used to determine the effect of long X-ray exposure. Finally, full-field strain from DVC was computed to quantify residual strain distribution at the cartilage-bone interface following unconfined compression test (ex situ). It was found that enhanced gray-scale feature recognition at the cartilage-bone interface was achieved using phase-contrast, resulting in reduced DVC strain uncertainties compared to absorption. Residual strains up to ~7000 µε in the articular cartilage were transferred to subchondral bone via the calcified cartilage and micromechanics revealed the predominant effect of long phase-contrast X-ray exposure in reducing both stiffness and hardness of the articular cartilage. The results of this study will pave the way for further development and refinement of the techniques, improving XCT-based strain measurements in cartilage-bone and other soft-hard tissue interfaces.

Dynamic shear behavior of mandibular condylar cartilage is dependent on testing direction

Little information is available on the direction-dependency of shear behavior in mandibular condylar cartilage. Therefore, we tested the hypothesis that such a dependency of the dynamic shear properties is present in mandibular condylar cartilage. From each of 17 condyles, two cartilage–bone plugs were dissected and tested in a simple shear sandwich configuration under a compressive strain of 10%. Sinusoidal shear strain (frequency range: 0.01–10 Hz) was applied in the medio-lateral or antero-posterior direction with an amplitude of 1.0%, 2.0%, and 3.0%. The magnitudes of the dynamic shear moduli, as calculated from the resulting shear stress, were found to increase with applied frequency and the shear strain amplitude. The values of ∣ G*∣, G′ and G″ for a medio-laterally applied shear were about 20–33% of those in the antero-posterior shear, although the loss tangent (elasticity/viscosity ratio) was almost the same. In conclusion, the present results clearly show the direction-dependent characteristic of the mandibular condylar cartilage in dynamic shear.

Biomechanical response of condylar cartilage‐on‐bone to dynamic shear

Shear stress can result in fatigue, damage, and irreversible deformation of the mandibular condylar cartilage. However, little information is available on its dynamic properties in shear. We tested the hypothesis that the dynamic shear properties of the condylar cartilage depend on the frequency and amplitude of shear strain. Ten porcine mandibular condyles were used for dynamic shear tests. Two cartilage-bone plugs were dissected from each condyle and tested in a simple shear sandwich configuration under a compressive strain of 10%. Sinusoidal shear strain was applied with an amplitude of 1.0, 2.0, and 3.0% and a frequency range between 0.01 and 10 Hz. The magnitudes of the shear dynamic moduli were found to be dependent on the frequency and the shear strain amplitude. They increased with shear strain. tan delta ranged from 0.2 to 0.4, which means that the cartilage is primarily elastic in nature and has a small but not negligible viscosity. In conclusion, the present results show that the shear behavior of the mandibular condylar cartilage is dependent on the frequency and amplitude of the applied shear strain. The observed shear characteristics suggest a significant role of shear strain on the interstitial fluid flow within the cartilage.

The effect of decalcification on the microstructure of articular cartilage assessed by 2H double quantum filtered spectroscopic MRI

2H quadrupolar splitting of deuterated water molecules is a sensitive measure of the order and density of the collagen fibers in articular cartilage. In the calcified zone, near the bone, two pairs of quadrupolar split satellites were previously observed. To examine whether the large splitting observed originates from the presence of calcium ions and hydroxyapatite, one-dimensional 2H single and double quantum filtered spectroscopic imaging were performed on articular cartilage-bone plugs before and after decalcification. After decalcification, the magnitude splitting of the two pairs of satellites did not change and orientation dependency was kept. However, the intensity of the large splitting was greatly enhanced. According to these results the two pairs of satellites do not stem from the presence of calcium ions and hydroxyapatite but originate from the presence of two groups of collagen fibers with different degrees of hydration. The enhanced intensity of the large splitting is attributed to an increased amount of water molecules that fill the void, resulting from the removal of hydroxyapatite, which resides near the fibers responsible for the large splitting. The quadrupolar splitting observed in the trabecular bone was not orientation-dependent, indicating a random orientation of the collagen fibers in that tissue.

T2 relaxation time measurements in osteoarthritis.

Quantification of changes in T(2) relaxation time, in human cartilage, with progression of osteoarthritis (OA), and evaluation of qualitative correlations with clinical evaluation, histology and polarized light microscopy (PLM). Cartilage-bone plugs were harvested from fresh cadaveric knees (n = 10) and specimens after surgical knee replacement (n = 2) at 12 locations, including lateral and medial sides of tibia, femora and patella. Magnetic resonance imaging was performed at 1.5 Tesla using a.2D spin echo sequence. Histological slices were assessed for OA severity through a grading scale based on combined histological and PLM results. T(2) values in clinically moderate OA were generally higher than in severe OA and normal cartilage. Significant association was established between normal and early OA subjects and T(2) variation, in the medial compartment of the knee (p < 0.05) but especially in the medial tibial cartilage (p < 0.00005). As expected, medial and lateral tibio-femoral compartments underwent more severe degeneration. Additionally, there were intracompartmental variation of the relaxation times and histological patterns, which demonstrate the underlying focal involvement of OA in the knee. Furthermore, T(2) values reflected OA pathogenesis with a positive correlation with histology grading scale. Finally, increased T(2) is correlated to histological degeneration of cartilage and may be a good marker for early OA in tibial articular cartilage.

Micro CT-based finite element analysis: an enhanced follow-up tool for bone remodelling around threaded implants

Objective: The objective of this study was to find a correlation between magnetic resonance (MR) appearance and biochemical composition of the normal articular cartilage by comparing the laminar aspects with the distribution of the two principal matrix components: proteoglycans and collagen.Design: T2-weighted MR microimages of porcine cartilage-bone plugs, excised from both the habitually loaded and habitually unloaded regions of the proximal end of the humerus, were obtained using a spin-echo sequence. Proteoglycans (PGs) were monitored by histology and by measuring the uronate and the sulfur content of the tissue; a histological method and the chemical determination of hydroxyproline were used for the evaluation of the collagen content.Results: The ‘loaded’ cartilage exhibited the expected MR laminar appearance whereas the ‘unloaded’ tissue appeared to be more homogeneous. The PG content in the ‘loaded’ cartilage, was found to be 2.4 times higher than in the habitually unloaded tissue, exhibiting an increasing trend from the articular surface to the bone. In the ‘unloaded’ cartilage the uronate distribution was more uniform with a higher concentration in the intermediate zone. The mean collagen content of both cartilage regions was found to be about 39% of the tissue dry weight. Histology and hydroxyproline distribution pattern showed that collagen was particularly concentrated at the surface and in a central zone of the ‘loaded’ cartilage whereas in the ‘unloaded’ tissue collagen was evident only at the surface. In accordance with the collagen distribution, transverse relaxation (T2) times in ‘loaded’ cartilage showed a minimum value at the articular surface and another minimum in a central region. On the contrary, the average T2value of the ‘unloaded’ tissue was high at the surface and decreased rapidly in the deeper zones.Conclusion: These results demonstrate that the MR appearance of articular cartilage correlates with the collagen content, but not with that of PGs, of the different zones. Other matrix components might, however, influence the MR appearance by contributing to the macromolecular organization of the tissue.

Heterogeneity of cartilage laminae in MR imaging.

The purpose of this study was to investigate the discrepancy in the number of laminae observed in magnetic resonance (MR) images of articular cartilage (the magic angle effect in MRI of cartilage). Microscopic MR imaging (muMRI) experiments were carried out at 14-micrometer pixel resolution on full-depth cartilage-bone plugs from several locations (central, intermediate, and peripheral) on the humeral heads of two young healthy beagles. When the articular surface of the plug was perpendicular to the direction of the magnetic field, the cartilage appeared to have two layers in the plugs from the central locations of the humeral head, three layers in the plugs from the greater tubercle side of the humeral head, and three or five layers in the plugs from the lesser tubercle side. This heterogeneity of cartilage laminae was observed within a single humeral head and was symmetrical about the median plane of the animal. This result suggests that some structural variations related to cartilage structure in various regions of load bearing may cause some unique laminar patterns seen in MRI of cartilage. This novel and new observation may resolve the controversy about whether cartilage appears as two or three layers in MR images. A comprehensive model for the collagen structure over a curved two-dimensional surface of a joint is suggested as a replacement of the classic three-zone model of fiber orientation in collagen. This heterogeneity of cartilage laminae is speculated to be related to the load-bearing status of the tissue in the joint. The ability to visualize such structural heterogeneity is important because of the direct connection between collagen structure and the mechanical characteristics of cartilage.

Heterogeneity of cartilage laminae in MR imaging

The purpose of this study was to investigate the discrepancy in the number of laminae observed in magnetic resonance (MR) images of articular cartilage (the magic angle effect in MRI of cartilage). Microscopic MR imaging (muMRI) experiments were carried out at 14-micrometer pixel resolution on full-depth cartilage-bone plugs from several locations (central, intermediate, and peripheral) on the humeral heads of two young healthy beagles. When the articular surface of the plug was perpendicular to the direction of the magnetic field, the cartilage appeared to have two layers in the plugs from the central locations of the humeral head, three layers in the plugs from the greater tubercle side of the humeral head, and three or five layers in the plugs from the lesser tubercle side. This heterogeneity of cartilage laminae was observed within a single humeral head and was symmetrical about the median plane of the animal. This result suggests that some structural variations related to cartilage structure in various regions of load bearing may cause some unique laminar patterns seen in MRI of cartilage. This novel and new observation may resolve the controversy about whether cartilage appears as two or three layers in MR images. A comprehensive model for the collagen structure over a curved two-dimensional surface of a joint is suggested as a replacement of the classic three-zone model of fiber orientation in collagen. This heterogeneity of cartilage laminae is speculated to be related to the load-bearing status of the tissue in the joint. The ability to visualize such structural heterogeneity is important because of the direct connection between collagen structure and the mechanical characteristics of cartilage.

MR-microscopic visualization of anisotropic internal cartilage structures using the magic angle technique.

NMR microscopic studies of articular cartilage at 7.1 T are presented. Using a special experimental design, T2-weighted spin-echo images of cartilage-bone plugs were taken under variable angles with respect to the static magnetic field B0 to visualize the angular-dependent representation of internal matrix structures mediated by the collagen network arrangement. To quantify the observed orientational effect in the MR images, exact measurements of the transverse relaxation time T2 were taken using the CPMG sequence. The NMR experiments show the strong influence of the cartilage orientation with respect to the static magnetic field on the inhomogeneous appearance of the articular cartilage in the MR image. Additionally performed polarization light microscopic investigations demonstrate the direct relation between the oriented collagenous structures and the anisotropic regions observed in the MR images. A simple cartilage matrix model derived from the experimental findings is proposed, and consequences for the clinical assessment of the articular joint are discussed.

Correlation between biochemical composition and magnetic resonance appearance of articular cartilage

Objective: The objective of this study was to find a correlation between magnetic resonance (MR) appearance and biochemical composition of the normal articular cartilage by comparing the laminar aspects with the distribution of the two principal matrix components: proteoglycans and collagen.Design: T2-weighted MR microimages of porcine cartilage-bone plugs, excised from both the habitually loaded and habitually unloaded regions of the proximal end of the humerus, were obtained using a spin-echo sequence. Proteoglycans (PGs) were monitored by histology and by measuring the uronate and the sulfur content of the tissue; a histological method and the chemical determination of hydroxyproline were used for the evaluation of the collagen content.Results: The ‘loaded’ cartilage exhibited the expected MR laminar appearance whereas the ‘unloaded’ tissue appeared to be more homogeneous. The PG content in the ‘loaded’ cartilage, was found to be 2.4 times higher than in the habitually unloaded tissue, exhibiting an increasing trend from the articular surface to the bone. In the ‘unloaded’ cartilage the uronate distribution was more uniform with a higher concentration in the intermediate zone. The mean collagen content of both cartilage regions was found to be about 39% of the tissue dry weight. Histology and hydroxyproline distribution pattern showed that collagen was particularly concentrated at the surface and in a central zone of the ‘loaded’ cartilage whereas in the ‘unloaded’ tissue collagen was evident only at the surface. In accordance with the collagen distribution, transverse relaxation (T2) times in ‘loaded’ cartilage showed a minimum value at the articular surface and another minimum in a central region. On the contrary, the average T2value of the ‘unloaded’ tissue was high at the surface and decreased rapidly in the deeper zones.Conclusion: These results demonstrate that the MR appearance of articular cartilage correlates with the collagen content, but not with that of PGs, of the different zones. Other matrix components might, however, influence the MR appearance by contributing to the macromolecular organization of the tissue.

Diffusion and relaxation mapping of cartilage-bone plugs and excised disks using microscopic magnetic resonance imaging.

Spatially resolved maps of proton self-diffusion coefficients (D) and relaxation times (T1 and T2) were obtained on cartilage-bone plug samples and on excised disks of canine cartilage at a transverse resolution of 30 microns, using microscopic magnetic resonance imaging (micro-MRI). Results are compared for excised disks of cartilage and intact cartilage-bone plugs. Correlations between the absolute water concentration, the self-diffusion coefficient and the T1 relaxation are reported. The diffusion coefficient is not a linear function of water concentration. The thickness of the disks is 600 microns, compared with the ca. 900 microns observed for the cartilage-bone plugs, presumably due to the absence of the interfacial or tidemark layer of interdigitated cartilage and bone in the former samples. Our results suggest that excised disks of cartilage are excellent models for the articular surface and the first 500 or so microns of tissue. The molecular parameters of spin-spin and spin-lattice relaxation times, as well as the water self-diffusion coefficient, are virtually identical in the two types of samples. However, the cartilage-bone plugs have the additional feature of permitting the study of the tidemark region, a region that likely plays a major role in the transmission of mechanical force.