Lori A. Setton

Compressive and shear properties of alginate gel: effects of sodium ions and alginate concentration.

The equilibrium and viscoelastic properties of alginate gel crosslinked with Ca2+ were determined as a function of alginate concentration and duration of exposure to physiologic concentrations of NaCl. Compressive and shear stress relaxation tests and oscillatory shear tests were performed to measure the material properties at two time periods after storage in NaCl compared to no NaCl exposure. The effect of concentration was determined by testing 1-3% alginate gel in a bath of physiological NaCl and CaCl2. After 15 h of exposure to NaCl, the compressive, equilibrium shear, and dynamic shear moduli decreased by 63, 84, and 90% of control values, respectively. The material properties exhibited no further changes after 7 days of exposure to NaCl. The loss angle and amplitude of the relaxation function in the shear also decreased, indicating less viscous behaviors in both dynamic and transient configurations. All moduli, but not the loss angle, significantly increased with increasing alginate concentration. The observed decrease in compressive and shear stiffness for alginate gel after exposure to Na+ was significant and indicated that physiological conditions will soften the gel over a time period of up to 7 days after gelation. The alginate gel retains significant solid-like behaviors, however, as measured by a loss angle of approximately 3 degrees. This study provides the first available data for material properties of alginate gel tested in physiological saline.

The biphasic poroviscoelastic behavior of articular cartilage: Role of the surface zone in governing the compressive behavior

Surface fibrillation of articular cartilage is an early sign of degenerative changes in the development of osteoarthritis. To assess the influence of the surface zone on the viscoelastic properties of cartilage under compressive loading, we prepared osteochondral plugs from skeletally mature steers, with and without the surface zone of articular cartilage, for study in the confined compression creep experiment. The relative contributions of two viscoelastic mechanisms, i.e. a flow-independent mechanism [Hayes and Bodine, J. Biomechanics 11, 407-419 (1978)], and a flow-dependent mechanism [Mow et al. J. biomech. Engng 102, 73-84 (1980)], to the compressive creep response of these two types of specimens were determined using the biphasic poroviscoelastic theory proposed by Mak. [J. Biomechanics 20, 703-714 (1986)]. From the experimental results and the biphasic poroviscoelastic theory, we found that frictional drag associated with interstitial fluid flow and fluid pressurization are the dominant mechanisms of load support in the intact specimens, i.e. the flow-dependent mechanisms alone were sufficient to describe normal articular cartilage compressive creep behavior. For specimens with the surface removed, we found an increased creep rate which was derived from an increased tissue permeability, as well as significant changes in the flow-independent parameters of the viscoelastic solid matrix. permeability, as well as significant changes in the flow-independent parameters of the viscoelastic solid matrix. From these tissue properties and the biphasic poroviscoelastic theory, we determined that the flow-dependent mechanisms of load support, i.e. frictional drag and fluid pressurization, were greatly diminished in cartilage without the articular surface. Calculations based upon these material parameters show that for specimens with the surface zone removed, the cartilage solid matrix became more highly loaded during the early stages of creep. This suggests that an important function of the articular surface is to provide for a low fluid permeability, and thereby serve to restrict fluid exudation and increase interstitial fluid pressurization. Thus, it is likely that with increasing severity of damage to the articular surface, load support in cartilage under compression shifts from the flow-dependent modes of fluid drag and pressurization to increased solid matrix stress. This suggests that it is important to maintain the integrity of the articular surface in preserving normal compressive behavior of the tissue and normal load carriage in the joint.

Degeneration affects the anisotropic and nonlinear behaviors of human anulus fibrosus in compression

Axial and radial specimens of non-degenerate and degenerate human anulus fibrosus (AF) were tested in confined compression to test the hypothesis that degeneration significantly affects the compressive properties of AF. Due to the highly oriented structure of AF, a secondary objective was to investigate anisotropic behaviors of AF in compression. Uniaxial swelling and stress relaxation experiments were performed on site-matched samples of anulus from the anterior outer region of L2-3 intervertebral discs. The experimental stress-relaxation behavior was modeled using the finite deformation biphasic theory and a finite-difference approximation scheme. Significant effects of degeneration but not orientation were detected for the reference stress offset, sigma(offset), and parameters describing the compressive stiffness (i.e. reference aggregate modulus, H(A0), and nonlinear stiffening coefficient, beta). Average values were 0.13+/-0.06 and 0.05+/-0.05 MPa for sigma(offset), 0.56+/-0.21 and 1.10+/-0.53 MPa for H(A0) and 2.13+/-1.48 and 0.44+/-0.61 for beta for all normal and degenerate specimens, respectively. No significant effect of degeneration or orientation were detected for either of the parameters describing the strain-dependent permeability (i.e. reference permeability, k0 and strain-dependent permeability coefficient, M) with average values for all specimens of 0.20+/-0.10 x 10(-15) m4/N-s and 1.18+/-1.30 for k0 and M, respectively. The loss of sigma(offset) was compensated with an elastic stiffening and change in the shape of the equilibrium stress-strain curve with H(A0) for degenerate tissues almost twice that of normal tissues and beta less than one sixth. The increase in reference elastic modulus with degeneration is likely related to an increase in tissue density resulting from the loss of water content. The significant effects of degeneration reported in this study suggested a shift in load carriage from fluid pressurization and swelling pressure to deformation of the solid matrix of the AF. The results also suggest that the highly organized and layered network of the anulus fibrosus, which gives rise to significant anisotropic effects in tension, does not play a major role in contributing to the magnitude of compressive stiffness or the mechanisms of fluid flow of the anulus in the confined compression configuration.

The Pericellular Matrix as a Transducer of Biomechanical and Biochemical Signals in Articular Cartilage

Abstract:  The pericellular matrix (PCM) is a narrow tissue region surrounding chondrocytes in articular cartilage, which together with the enclosed cell(s) has been termed the “chondron.” While the function of this region is not fully understood, it is hypothesized to have important biological and biomechanical functions. In this article, we review a number of studies that have investigated the structure, composition, mechanical properties, and biomechanical role of the chondrocyte PCM. This region has been shown to be rich in proteoglycans (e.g., aggrecan, hyaluronan, and decorin), collagen (types II, VI, and IX), and fibronectin, but is defined primarily by the presence of type VI collagen as compared to the extracellular matrix (ECM). Direct measures of PCM properties via micropipette aspiration of isolated chondrons have shown that the PCM has distinct mechanical properties as compared to the cell or ECM. A number of theoretical and experimental studies suggest that the PCM plays an important role in regulating the microenvironment of the chondrocyte. Parametric studies of cell–matrix interactions suggest that the presence of the PCM significantly affects the micromechanical environment of the chondrocyte in a zone‐dependent manner. These findings provide support for a potential biomechanical function of the chondrocyte PCM, and furthermore, suggest that changes in the PCM and ECM properties that occur with osteoarthritis may significantly alter the stress‐strain and fluid environments of the chondrocytes. An improved understanding of the structure and function of the PCM may provide new insights into the mechanisms that regulate chondrocyte physiology in health and disease.

Degeneration and aging affect the tensile behavior of human lumbar anulus fibrosus.

Studyb Design Samples of human lumbar (L3-L4) anulus fibrosus from four different anatomic sites (anterior outer, posterolateral outer, anterior inner, posterolateral inner), ranging from normal to severely degenerate, were studied in uniaxial tension and measured for water content. Objectives To evaluate the effects of aging and degeneration on the tensile properties and hydration of the anulus fibrosus in a site-specific manner, The relationship between hydration and parameters of the tenaile behavior were investigated. Summary of Background Data Degeneration and aging have been shown to be related to dramatic changes in the composition and structure of the anulus fibrosus. The associated changes in the tensile, compressive, and shear properties of the anulus fibrosus have not been documented. Numerical studies using finite element models have attempted to simulate the degenerative process by incorporating estimated mechanical properties meant to represent the degenerate anulus fibrosus. Their results present findings that suggest that altered material properties of the anulus fibrosus affect the mechanics of the entire intervertebral disc. Methods Samples of human lumbar anulus fibrosus were classified by grade of degeneration based on a morphologic grading scheme. Multiple layer anulus specimens from four sites in the disc were tested in uniaxial tension under quasistatic conditions in a physiologic saline bath. The tensile modulus, prission;s ratio failure stress and strain, the strain energy density to failure, and the corresponding hydration were determined for each test sample. Result The possons ratio, Failure stress, and strain energy density of the anulus fibrosus were found to be affected significantly by degeneration, with some evldence of a sensitivity of the tensile modulus to gradeof degeneration. All nmaterial properties were found to exhibit a significant and greater dependence on site within the disc than on degenerative grade. Weak corelations between aging and the Poissons ratio and strain energy density were observed. Water content of anulus fibrosus tissue was not affected by degeneration or again, although correlations with tensile properties fwere observed. Conclusions The dramatic changes in morphology, composition, and structure that occur in anulus fibrosus with agin and degeneration are accompanied by specific variations in the tensile properties, Which were generally small in magnitude. Position of the anulus fibrosus within the intertebral disc, particularly in the radial direction, appeared to be the most important variable affecting anulus fibrosus tensile properties. This dependence on position did not change with either aging or degeneration. Results from the present study may be useful in future finite element models to assess how altered material properties of the anulus fibrosus during degeneration and aging may affect the mechanics of the entire intervertebral disc.

Anisotropic and inhomogeneous tensile behavior of the human anulus fibrosus: experimental measurement and material model predictions.

The anulus fibrosus (AF) of the intervertebral disc exhibits spatial variations in structure and composition that give rise to both anisotropy and inhomogeneity in its material behaviors in tension. In this study, the tensile moduli and Poissons ratios were measured in samples of human AF along circumferential, axial, and radial directions at inner and outer sites. There was evidence of significant inhomogeneity in the linear-region circumferential tensile modulus (17.4+/-14.3 MPa versus 5.6+/-4.7 MPa, outer versus inner sites) and the Poissons ratio v21 (0.67+/-0.22 versus 1.6+/-0.7, outer versus inner), but not in the axial modulus (0.8+/-0.9 MPa) or the Poissons ratios V12 (1.8+/-1.4) or v13 (0.6+/-0.7). These properties were implemented in a linear an isotropic material model of the AF to determine a complete set of model properties and to predict material behaviors for the AF under idealized kinematic states. These predictions demonstrate that interactions between fiber populations in the multilamellae AF significantly contribute to the material behavior, suggesting that a model for th

Tensile properties of nondegenerate human lumbar anulus fibrosus

Study Design The in vitro tensile behavior of multiple‐layer samples of anulus fibrosus were investigated from nondegenerate intervertebral discs. Objectives To quantify the intrinsic tensile behavior of nondegenerate anulus fibrosus and the variations with position and age in the intervertebral disc. Summary of Background Data Tension is an important loading mode in the anulus fibrosus. The tensile behavior of single‐ and multiple‐layer samples of anulus fibrosus has been shown to vary with specimen orientation, position in the disc, and environmental conditions. Little is known of the changes in these site‐specific tensile properties of the anulus with aging or degeneration of the intervertebral disc. Methods Multiple‐layer specimens of anulus fibrosus were harvested with an orientation parallel to the circumference of the disc. Constant strain rate and uniaxial tensile tests were performed in 0.15 mol/l NaCl at slow strain rates to measure the intrinsic properties of the collagen‐proteoglycan matrix of the anulus fibrosus. The tensile modulus, failure stress, failure strain, and strain energy density were determined. Statistical analyses were done to evaluate regional and age‐related differences in these properties. Results Significant radial and circumferential variations in the intrinsic tensile properties of anular samples were detected. The anterior anulus fibrosus had larger values for tensile moduli and failure stresses than the posterolateral anulus. Also, the outer regions of the anulus had greater moduli and failure stresses and lower failure strains than the inner regions. Strain energy density did not vary significantly with region. Significant, but very weak, correlations were detected between tensile properties and age of the intervertebral disc. Conclusions The observed variations in tensile behavior of multiple‐layer anulus samples indicate that larger variations in tensile modulus and failure properties occur with radial position in the disc than from anterior to posterolateral regions. This pattern is likely related to site‐specific variations in the tensile properties of the single‐layer samples of anulus fibrosus lamellae and the organization of successive lamellae and their interactions. The results of the present study suggest that factors other than age, such as compositional and structural variations in the disc, are the most important determinants of tensile behavior of the anulus fibrosus.

Photocrosslinkable hyaluronan as a scaffold for articular cartilage repair.

Hyaluronan-based scaffolds are of interest for tissue-engineered cartilage repair due to an important role for hyaluronan in cartilage development and function. In this study, an in situ photocrosslinkable hyaluronan (HA-MA) was developed and evaluated as a scaffold for articular cartilage repair. Chondrocytes were encapsulated in crosslinked HA-MA and evaluated for their ability to synthesize cartilaginous matrix in vitro. The mechanical and physical properties of the crosslinked HA-MA hydrogels were similar to that of other hydrogels, with compressive and dynamic shear moduli of 0.6 and 0.3 kPa, respectively, and diffusion coefficients of 600–8000 μm2/s depending on molecular weight. Chondrocytes remained rounded in the HA-MA hydrogels in vitro, and accumulated significant amounts of cartilaginous matrix. Osteochondral defects filled with HA-MA were infiltrated with cells, appeared to integrate well with native tissue, and also accumulated substantial cartilaginous matrix by 2 weeks after surgery. In summary, photocrosslinkable HA-MA promoted the retention of the chondrocytic phenotype and cartilage matrix synthesis for encapsulated chondrocytes in vitro and accelerated healing in an in vivo osteochondral defect model.

Compressive mechanical properties of the human anulus fibrosus and their relationship to biochemical composition.

To enhance understanding of the biomechanical role of the intervertebral disc, the compressive properties and biochemical composition of nondegenerate samples of anulus fibrosus were determined as a function of radial position, region, and level. Because of the large swelling propensity of this tissue, a method was developed to test excised specimens while maintaining their in situ geometry and hydration. Using an analysis based on linear biphasic theory, the compressive modulus, hydraulic permeability, and isometric swelling pressure of the anulus fibrosus were determined and correlated with the tissue composition. The findings indicate that the anulus fibrosus is inhomogeneous, with regional and radial variations in both material properties and biochemical composition. The results of this study suggest that both structural and compositional factors may determine the mechanical behavior.

Proinflammatory cytokine expression profile in degenerated and herniated human intervertebral disc tissues

OBJECTIVE Prior reports document macrophage and lymphocyte infiltration with proinflammatory cytokine expression in pathologic intervertebral disc (IVD) tissues. Nevertheless, the role of the Th17 lymphocyte lineage in mediating disc disease remains uninvestigated. We undertook this study to evaluate the immunophenotype of pathologic IVD specimens, including interleukin-17 (IL-17) expression, from surgically obtained IVD tissue and from nondegenerated autopsy control tissue. METHODS Surgical IVD tissues were procured from patients with degenerative disc disease (n = 25) or herniated IVDs (n = 12); nondegenerated autopsy control tissue was also obtained (n = 8) from the anulus fibrosus and nucleus pulposus regions. Immunohistochemistry was performed for cell surface antigens (CD68 for macrophages, CD4 for lymphocytes) and various cytokines, with differences in cellularity and target immunoreactivity scores analyzed between surgical tissue groups and between autopsy control tissue regions. RESULTS Immunoreactivity for IL-4, IL-6, IL-12, and interferon-gamma (IFNgamma) was modest in surgical IVD tissue, although expression was higher in herniated IVD samples and virtually nonexistent in control samples. The Th17 lymphocyte product IL-17 was present in >70% of surgical tissue fields, and among control samples was detected rarely in anulus fibrosus regions and modestly in nucleus pulposus regions. Macrophages were prevalent in surgical tissues, particularly herniated IVD samples, and lymphocytes were expectedly scarce. Control tissue revealed lesser infiltration by macrophages and a near absence of lymphocytes. CONCLUSION Greater IFNgamma positivity, macrophage presence, and cellularity in herniated IVDs suggests a pattern of Th1 lymphocyte activation in this pathology. Remarkable pathologic IVD tissue expression of IL-17 is a novel finding that contrasts markedly with low levels of IL-17 in autopsy control tissue. These findings suggest involvement of Th17 lymphocytes in the pathomechanism of disc degeneration.