Direct measurement of osmotic pressure of glycosaminoglycan solutions by membrane osmometry at room temperature

Articular cartilage is a hydrated soft tissue composed of negatively charged proteoglycans fixed within a collagen matrix. This charge gradient causes the tissue to imbibe water and swell, creating a net osmotic pressure that enhances the tissue's ability to bear load. In this study we designed and utilized an apparatus for directly measuring the osmotic pressure of chondroitin sulfate, the primary glycosaminoglycan found in articular cartilage, in solution with varying bathing ionic strength (0.015 M, 0.15 M, 0.5 M, 1 M, and 2 M NaCl) at room temperature. The osmotic pressure (pi) was found to increase nonlinearly with increasing chondroitin sulfate concentration and decreasing NaCl ionic bath environment. Above 1 M NaCl, pi changes negligibly with further increases in salt concentration, suggesting that Donnan osmotic pressure is negligible above this threshold, and the resulting pressure is attributed to configurational entropy. Results of the current study were also used to estimate the contribution of osmotic pressure to the stiffness of cartilage based on theoretical and experimental considerations. Our findings indicate that the osmotic pressure resulting from configurational entropy is much smaller in cartilage (based on an earlier study on bovine articular cartilage) than in free solution. The rate of change of osmotic pressure with compressive strain is found to contribute approximately one-third of the compressive modulus (H(A)(eff)) of cartilage (Pi approximately H(A)(eff)/3), with the balance contributed by the intrinsic structural modulus of the solid matrix (i.e., H(A) approximately 2H(A)(eff)/3). A strong dependence of this intrinsic modulus on salt concentration was found; therefore, it appears that proteoglycans contribute structurally to the magnitude of H(A), in a manner independent of osmotic pressure.     

A microstructural model of elastostatic properties of articular cartilage in confined compression

A microstructural model of cartilage was developed to investigate the relative contribution of tissue matrix components to its elastostatic properties. Cartilage was depicted as a tensed collagen lattice pressurized by the Donnan osmotic swelling pressure of proteoglycans. As a first step in modeling the collagen lattice, two-dimensional networks of tensed, elastic, interconnected cables were studied as conceptual models. The models were subjected to the boundary conditions of confined compression and stress-strain curves and elastic moduli were obtained as a function of a two-dimensional equivalent of swelling pressure. Model predictions were compared to equilibrium confined compression moduli of calf cartilage obtained at different bath concentrations ranging from 0.01 to 0.50 M NaCl. It was found that a triangular cable network provided the most consistent correspondence to the experimental data. The model showed that the cartilage collagen network remained tensed under large confined compression strains and could therefore support shear stress. The model also predicted that the elastic moduli increased with increasing swelling pressure in a manner qualitatively similar to experimental observations. Although the model did not preclude potential contributions of other tissue components and mechanisms, the consistency of model predictions with experimental observations suggests that the cartilage collagen network, prestressed by proteoglycan swelling pressure, plays an important role in supporting compression.     

Confined compression and torsion experiments on a pHEMA gel in various bath concentrations

The constitutive behaviour of cartilaginous tissue is the result of complex interaction between electrical, chemical and mechanical forces. Electrostatic interactions between fixed charges and mobile ions are usually accounted for by means of Donnan osmotic pressure. Recent experimental data show, however, that the shear modulus of articular cartilage depends on ionic concentration even if the strain is kept constant. Poisson–Boltzmann simulations suggest that this dependence is intrinsic to the double-layer around the proteoglycan chains. In order to verify this premise, this study measures whether—at a given strain—this ionic concentration–dependent shear modulus is present in a polymerized hydroxy-ethyl-methacrylate gel or not. A combined 1D confined compression and torque experiment is performed on a thin cylindrical hydrogel sample, which is brought in equilibrium with, respectively, 1, 0.1 and 0.03 M NaCl. The sample was placed in a chamber that consists of a stainless steel ring placed on a sintered glass filter, and on top a sintered glass piston. Stepwise ionic loading was cascaded by stepwise 1D compression, measuring the total stress after equilibration of the sample. In addition, a torque experiment was interweaved by applying a harmonic angular displacement and measuring the torque, revealing the relation between aggregate shear modulus and salt concentration at a given strain.     

Elastic,permeability and swelling properties of human intervertebral disc tissues: A benchmark for tissue engineering

The aim of functional tissue engineering is to repair and replace tissues that have a biomechanical function, i.e., connective orthopaedic tissues. To do this, it is necessary to have accurate benchmarks for the elastic, permeability, and swelling (i.e., biphasic-swelling) properties of native tissues. However, in the case of the intervertebral disc, the biphasic-swelling properties of individual tissues reported in the literature exhibit great variation and even span several orders of magnitude. This variation is probably caused by differences in the testing protocols and the constitutive models used to analyze the data. Therefore, the objective of this study was to measure the human lumbar disc annulus fibrosus (AF), nucleus pulposus (NP), and cartilaginous endplates (CEP) biphasic-swelling properties using a consistent experimental protocol and analyses. The testing protocol was composed of a swelling period followed by multiple confined compression ramps. To analyze the confined compression data, the tissues were modeled using a biphasic-swelling model, which augments the standard biphasic model through the addition of a deformation-dependent osmotic pressure term. This model allows considering the swelling deformations and the contribution of osmotic pressure in the analysis of the experimental data. The swelling stretch was not different between the disc regions (AF: 1.28±0.16; NP: 1.73±0.74; CEP: 1.29±0.26), with a total average of 1.42. The aggregate modulus (Ha) of the extra-fibrillar matrix was higher in the CEP (390 kPa) compared to the NP (100 kPa) or AF (30 kPa). The permeability was very different across tissue regions, with the AF permeability (64 E⁻¹⁶ m⁴/N s) higher than the NP and CEP (~5.5 E⁻¹⁶ m⁴/N s). Additionally, a normalized time-constant (3000 s) for the stress relaxation was similar for all the disc tissues. The properties measured in this study are important as benchmarks for tissue engineering and for modeling the disc's mechanical behavior and transport.     

Confined compression experiments on bovine nucleus pulposus and annulus fibrosus: sensitivity of the experiment in the determination of compressive modulus and hydraulic permeability

The biphasic material properties for nucleus pulposus tissue in confined compression have not been reported previously, and are required for a better understanding of intervertebral disc function and to provide material properties for use in finite-element models. The aims of this study were to determine linear and non-linear material properties for nucleus pulposus and annulus fibrosus tissues in confined compression, to define the influence of swelling conditions on these properties, and to determine the changes in the compressive modulus and hydraulic permeability induced by the repetition of the stress-relaxation experiment after a return to swelling pressure equilibrium. Specimens from caudal bovine nucleus and annulus were tested in confined compression stress-relaxation experiments and analyzed to quantify the compressive modulus and hydraulic permeability using linear and non-linear biphasic models. Our results suggested the use of confined swelling pre-test condition and non-linear biphasic model, which provided the material parameters with lowest relative variance and water content most representative of physiological conditions. Smaller compressive modulus and higher hydraulic permeability were obtained for the nucleus (H(A0)=0.31+/-0.04 MPa, k(0)=0.67+/-0.09 x 10(-15)m(4)/Ns) than for the annulus (H(A0)=0.74+/-0.13 MPa, k(0)=0.23+/-0.19 x 10(-15)m(4)/Ns), with relatively weak non-linearities. Strains up to 20% resulted in material properties that were significantly altered upon retesting. These altered material properties are an effort to quantify non-recoverable damage that occurs in disc tissue and suggest that in vivo exposure of disc tissues to low strain-rate and high-deformation loading conditions which outpace biological repair may result in altered mechanical behaviors.     

Anisotropic strain-dependent material properties of bovine articular cartilage in the transitional range from tension to compression

Articular cartilage exhibits complex mechanical properties such as anisotropy, inhomogeneity and tension-compression nonlinearity. This study proposes and demonstrates that the application of compressive loading in the presence of osmotic swelling can be used to acquire a spectrum of incremental cartilage moduli (EYi) and Poisson's ratios (upsilon ij) from tension to compression. Furthermore, the anisotropy of the tissue can be characterized in both tension and compression by conducting these experiments along three mutually perpendicular loading directions: parallel to split-line (1-direction), perpendicular to split-line (2-direction) and along the depth direction (3-direction, perpendicular to articular surface), accounting for tissue inhomogeneity between the surface and deep layers in the latter direction. Tensile moduli were found to be strain-dependent while compressive moduli were nearly constant. The peak tensile (+) Young's moduli in 0.15M NaCl were E+Y1=3.1+/-2.3, E+Y2=1.3+/-0.3, E+Y3(Surface)=0.65+/-0.29 and E+Y3(Deep)=2.1+/-1.2 MPa. The corresponding compressive (-) Young's moduli were E-Y1=0.23+/-0.07, E-Y2=0.22+/-0.07, E-Y3(Surface)=0.18+/-0.07 and E-Y3(Deep)=0.35+/-0.11 MPa. Peak tensile Poisson's ratios were upsilon+12=0.22+/-0.06, upsilon+21=0.13+/-0.07, upsilon+31(Surface)=0.10+/-0.03 and upsilon+31(Deep)=0.20+/-0.05 while compressive Poisson's ratios were upsilon-12=0.027+/-0.012, upsilon-21=0.017+/-0.07, upsilon-31(Surface)=0.034+/-0.009 and upsilon-31(Deep)=0.065+/-0.024. Similar measurements were also performed at 0.015 M and 2 M NaCl, showing strong variations with ionic strength. Results indicate that (a) a smooth transition occurs in the stress-strain and modulus-strain responses between the tensile and compressive regimes, and (b) cartilage exhibits orthotropic symmetry within the framework of tension-compression nonlinearity. The strain-softening behavior of cartilage (the initial decrease in EYi with increasing compressive strain) can be interpreted in the context of osmotic swelling and tension-compression nonlinearity.     

Swelling and de-swelling kinetics of gelatin hydrogels in ethanol-water marginal solvent

Controlled osmotic swelling and de-swelling measurements have been performed on gelatin, a polyampholyte, hydrogels suspended in water-ethanol marginal solvent at room temperature (20 degrees C) where the alcohol concentration was changed from 0 to 100% (v/v). The change in gel mass was monitored as function of time until osmotic equilibrium was established with the surrounding solvent. It was observed that osmotic pressure of polymer-solvent mixing, pi(m)相似文献   

Compression loading in vitro regulates proteoglycan synthesis by tendon fibrocartilage.

The regulation of proteoglycan synthesis in a fibrocartilaginous tissue by mechanical loading was assessed in vitro. Discs of bovine tendon fibrocartilage were loaded daily with unconfined, cyclic, uniaxial compression (5 s/min, 20 min/day) and the synthesis of large and small proteoglycans was measured by incorporation of [35S]sulfate. All discs synthesized predominantly large proteoglycan when first placed in culture. After 2 weeks in culture nonloaded discs synthesized predominantly small proteoglycans whereas loaded discs continued to produce predominantly large proteoglycan. The turnover of 35S-labeled proteoglycan was not significantly altered by the compression regime. Increased synthesis of large proteoglycans was induced by a 4-day compression regime following 21 days of culture without compression. Inclusion of cytochalasin B during compression mimicked this induction. Autoradiography demonstrated that cell proliferation was minimal and confined to the disc edges whereas 35S-labeled proteoglycan synthesis occurred throughout the discs. These experiments demonstrate that mechanical compression can regulate synthesis of distinct proteoglycan types in fibrocartilage.     

The osmotic pressure of chondroitin sulphate solutions: experimental measurements and theoretical analysis

We used equilibrium dialysis to measure the osmotic pressure of chondroitin sulphate (CS) solutions as a function of their concentration and fixed charge density (FCD) and the ionic strength and composition of the solution. Osmotic pressure varied nonlinearly with the concentration of chondroitin sulphate and in 0.15 M NaCl at FCDs typical of uncompressed cartilage (approximately 0.4 mmol/g extrafibrillar H2O) was approximately 3 atmospheres. Osmotic pressure fell by 60% as solution ionic strength increased up to about 1 M, but remained relatively constant at higher ionic strengths. The ratio of Ca2+ to Na+ in the medium was a minor determinant of osmotic pressure. The data are compared with a theoretical model of the electrostatic contribution to osmotic pressure calculated from the Poisson-Boltzmann equation using a rod-in-cell model for CS. The effective radius of the polyelectrolyte rod is taken as a free parameter. The model qualitatively reproduces the non-linear concentration dependence, but underestimates the osmotic pressure by an amount that is independent of ionic strength. This difference, presumably arising from oncotic and entropic effects, is approximately 1/3 of the total osmotic pressure at physiological polymer concentrations and ionic strength.     

Three-dimensional inhomogeneous triphasic finite-element analysis of physical signals and solute transport in human intervertebral disc under axial compression

A 3D inhomogeneous finite-element model for charged hydrated soft tissues containing charged/uncharged solutes was developed and applied to analyze the mechanical, chemical, and electrical signals within the human intervertebral disc during an axial unconfined compression. The effects of tissue properties and boundary conditions on the physical signals and the transport of fluid and solute were investigated. The numerical simulation showed that, during disc compression, the fluid pressurization and the effective (von Misses) solid stress were more pronounced in the annulus fibrosus (AF) region near the interface between AF and nucleus pulposus (NP). In NP, the distributions of the fluid pressure, effective stress, and electrical potential were more uniform than those in AF. The electrical signals were very sensitive to fixed charge density. Changes in material properties of NP (water content, fixed charge density, and modulus) affected fluid pressure, electrical potential, effective stress, and solute transport in the disc. This study is important for understanding disc biomechanics, disc nutrition, and disc mechanobiology.     

Protective effect of sucrose and sodium chloride for Lactococcus lactis during sublethal and lethal high-pressure treatments

The bactericidal effect of hydrostatic pressure is reduced when bacteria are suspended in media with high osmolarity. To elucidate mechanisms responsible for the baroprotective effect of ionic and nonionic solutes, Lactococcus lactis was treated with pressures ranging from 200 to 600 MPa in a low-osmolarity buffer or with buffer containing 0.5 M sucrose or 4 M NaCl. Pressure-treated cells were characterized in order to determine viability, the transmembrane difference in pH (DeltapH), and multiple-drug-resistance (MDR) transport activity. Furthermore, pressure effects on the intracellular pH and the fluidity of the membrane were determined during pressure treatment. In the presence of external sucrose and NaCl, high intracellular levels of sucrose and lactose, respectively, were accumulated by L. lactis; 4 M NaCl and, to a lesser extent, 0.5 M sucrose provided protection against pressure-induced cell death. The transmembrane DeltapH was reversibly dissipated during pressure treatment in any buffer system. Sucrose but not NaCl prevented the irreversible inactivation of enzymes involved in pH homeostasis and MDR transport activity. In the presence 0.5 M sucrose or 4 M NaCl, the fluidity of the cytoplasmic membrane was maintained even at low temperatures and high pressure. These results indicate that disaccharides protect microorganisms against pressure-induced inactivation of vital cellular components. The protective effect of ionic solutes relies on the intracellular accumulation of compatible solutes as a response to the osmotic stress. Thus, ionic solutes provide only asymmetric protection, and baroprotection with ionic solutes requires higher concentrations of the osmolytes than of disaccharides.     

Protective Effect of Sucrose and Sodium Chloride for Lactococcus lactis during Sublethal and Lethal High-Pressure Treatments

The bactericidal effect of hydrostatic pressure is reduced when bacteria are suspended in media with high osmolarity. To elucidate mechanisms responsible for the baroprotective effect of ionic and nonionic solutes, Lactococcus lactis was treated with pressures ranging from 200 to 600 MPa in a low-osmolarity buffer or with buffer containing 0.5 M sucrose or 4 M NaCl. Pressure-treated cells were characterized in order to determine viability, the transmembrane difference in pH (ΔpH), and multiple-drug-resistance (MDR) transport activity. Furthermore, pressure effects on the intracellular pH and the fluidity of the membrane were determined during pressure treatment. In the presence of external sucrose and NaCl, high intracellular levels of sucrose and lactose, respectively, were accumulated by L. lactis; 4 M NaCl and, to a lesser extent, 0.5 M sucrose provided protection against pressure-induced cell death. The transmembrane ΔpH was reversibly dissipated during pressure treatment in any buffer system. Sucrose but not NaCl prevented the irreversible inactivation of enzymes involved in pH homeostasis and MDR transport activity. In the presence 0.5 M sucrose or 4 M NaCl, the fluidity of the cytoplasmic membrane was maintained even at low temperatures and high pressure. These results indicate that disaccharides protect microorganisms against pressure-induced inactivation of vital cellular components. The protective effect of ionic solutes relies on the intracellular accumulation of compatible solutes as a response to the osmotic stress. Thus, ionic solutes provide only asymmetric protection, and baroprotection with ionic solutes requires higher concentrations of the osmolytes than of disaccharides.     

Osmotic pressure of DNA solutions and effective diameter of the double helix

A simple osmometer with nuclear filters (polymer films with pores of a preset diameter) were used to measure the osmotic pressure of Col E1 plasmid DNA solutions in the concentration range of 1-4 mg/ml DNA. Linear and open circular DNA forms proved to have the same osmotic pressure within the experimental accuracy. The results of the measurements were used for calculating the second virial coefficient A2 of the solution of DNA segments and the effective chain diameter d eff in the ionic strength range of 10(-2)-0.1 M. As the ionic strength is lowered from 0.1 to 10(-2) M the effective diameter of DNA increases from 80 to 220 A. The results are in rather good agreement with theory and with other experimental data.     

The influence of the fixed negative charges on mechanical and electrical behaviors of articular cartilage under unconfined compression

Unconfined compression test has been frequently used to study the mechanical behaviors of articular cartilage, both theoretically and experimentally. It has also been used in explant and gel-cell-complex studies in tissue engineering. In biphasic and poroelastic theories, the effect of charges fixed on the proteoglycan macromolecules in articular cartilage is embodied in the apparent compressive Young's modulus and the apparent Poisson's ratio of the tissue, and the fluid pressure is considered to be the portion above the osmotic pressure. In order to understand how proteoglycan fixed charges might affect the mechanical behaviors of articular cartilage, and in order to predict the osmotic pressure and electric fields inside the tissue in this experimental configuration, it is necessary to use a model that explicitly takes into account the charged nature of the tissue and the flow of ions within its porous interstices. In this paper, we used a finite element model based on the triphasic theory to study how fixed charges in the porous-permeable soft tissue can modulate its mechanical and electrochemical responses under a step displacement in unconfined compression. The results from finite element calculations showed that: 1) A charged tissue always supports a larger load than an uncharged tissue of the same intrinsic elastic moduli. 2) The apparent Young's modulus (the ratio of the equilibrium axial stress to the axial strain) is always greater than the intrinsic Young's modulus of an uncharged tissue. 3) The apparent Poisson's ratio (the negative ratio of the lateral strain to the axial strain) is always larger than the intrinsic Poisson's ratio of an uncharged tissue. 4) Load support derives from three sources: intrinsic matrix stiffness, hydraulic pressure and osmotic pressure. Under the unconfined compression, the Donnan osmotic pressure can constitute between 13%-22% of the total load support at equilibrium. 5) During the stress-relaxation process following the initial instant of loading, the diffusion potential (due to the gradient of the fixed charge density and the associated gradient of ion concentrations) and the streaming potential (due to fluid convection) compete against each other. Within the physiological range of material parameters, the polarity of the electric potential depends on both the mechanical properties and the fixed charge density (FCD) of the tissue. For softer tissues, the diffusion effects dominate the electromechanical response, while for stiffer tissues, the streaming potential dominates this response. 6) Fixed charges do not affect the instantaneous strain field relative to the initial equilibrium state. However, there is a sudden increase in the fluid pressure above the initial equilibrium osmotic pressure. These new findings are relevant and necessary for the understanding of cartilage mechanics, cartilage biosynthesis, electromechanical signal transduction by chondrocytes, and tissue engineering.     

Validation and application of an intervertebral disc finite element model utilizing independently constructed tissue-level constitutive formulations that are nonlinear,anisotropic, and time-dependent

Finite element (FE) models are advantageous in the study of intervertebral disc mechanics as the stress–strain distributions can be determined throughout the tissue and the applied loading and material properties can be controlled and modified. However, the complicated nature of the disc presents a challenge in developing an accurate and predictive disc model, which has led to limitations in FE geometry, material constitutive models and properties, and model validation. The objective of this study was to develop a new FE model of the intervertebral disc, to validate the model?s nonlinear and time-dependent responses without tuning or calibration, and to evaluate the effect of changes in nucleus pulposus (NP), cartilaginous endplate (CEP), and annulus fibrosus (AF) material properties on the disc mechanical response. The new FE disc model utilized an analytically-based geometry. The model was created from the mean shape of human L4/L5 discs, measured from high-resolution 3D MR images and averaged using signed distance functions. Structural hyperelastic constitutive models were used in conjunction with biphasic-swelling theory to obtain material properties from recent tissue tests in confined compression and uniaxial tension. The FE disc model predictions fit within the experimental range (mean±95% confidence interval) of the disc?s nonlinear response for compressive slow loading ramp, creep, and stress-relaxation simulations. Changes in NP and CEP properties affected the neutral-zone displacement but had little effect on the final stiffness during slow-ramp compression loading. These results highlight the need to validate FE models using the disc?s full nonlinear response in multiple loading scenarios.     

Influence of a stationary magnetic field on water relations in lettuce seeds. Part I: theoretical considerations.

The theoretical calculation about the dependence of the ionic current density across the cellular membrane on the intensity of the magnetic field applied to cellular tissue is presented. This interaction induces changes in the magnitude of the ionic current density across the cellular membrane and in the ionic concentration, and it also causes alterations in the osmotic pressure and in the capacity of the cellular tissues to absorb water. The magnetic field dependence of the ionic current densities J(p) (B) (positive ions) and J(n) (B) (negative ions), the membrane conductivity sigma (B), the ionic concentration in both membrane sides c(B), the osmotic pressure pi (B), and the water uptake rate by seeds k(w) (B) are presented. The increase in water uptake rate due to the applied magnetic field may be the explanation of the recently reported increase in the germination speed of the seeds treated with stationary magnetic fields.     

Heterogeneity of heparan sulfate chains in a proteoglycan from bovine lung

A heparan sulfate proteoglycan from bovine lung gas-exchange tissue was isolated by extraction of the tissue with 4.0 M guanidine HCl in the presence of multiple protein inhibitors. The proteoglycan was purified by precipitation with cetylpyridinium chloride in 0.5 M KCl followed by CsCl isopycnic centrifugation (po = 1.45) in 4.0 M guanidine/HCl. Further purification was achieved by gel filtration on Sepharose CL-2B and by chromatography in DEAE-Sepharose CL-6B column. The proteoglycan had 14.9% protein and 22.4% uronate. Heparan sulfate chains from the proteoglycan were isolated after beta-elimination. Fractionation of heparan sulfate chains was achieved on Dowex-1 Cl- column, eluting with a stepwise increase in the concentration of NaCl, 1.0 to 2.0 M with 0.2 M increments. Of the total heparan sulfate recovered from the column, about 10% eluted by 1.2 M NaCl, 68% by 1.4 M NaCl, 18% by 1.6 M NaCl and 4% by 1.8 M NaCl. The fractions varied in their total and N-sulfate ester contents and iduronic acid to glucuronic acid ratios. The fraction that eluted from the Dowex-1 Cl- column at 1.6 M NaCl had the highest molecular weight, 37000, and the fraction that eluted at 1.8 M NaCl had the lowest molecular weight, 12000, as determined by gel filtration method, and the greatest sulfate content. The core protein, obtained by digestion of proteoglycan by heparan sulfate lyase, showed mostly a single band in SDS-polyacrylamide gel electrophoresis. The observations indicate a heterogeneity of the composition of heparan sulfate chains in the proteoglycan. This heterogeneity likely contributes to variations in biologic properties of different heparan sulfate proteoglycan preparations.     

A mathematical model of proximal tubule absorption

A previous model of the mechanisms of flow through epithelia was modified and extended to include hydrostatic and osmotic pressures in the cells and in the peritubular capillaries. The differential equations for flow and concentration in each region of the proximal tubule were derived. The equations were solved numerically by a finite difference method. The principal conclusions are: (i) Cell NaCl concentration remains essentially isotonic over the pressure variations considered; (ii) channel NaCl concentration varies only a few mosmol from isotonicity, and the hydrostatic and osmotic pressure differences across the cell wall are of the same order of magnitude; (iii) both reabsorbate osmolality and pressure-induced flow are relatively insensitive to the geometry of the system; (iv) a strong equilibrating mechanism exists in the sensitivity of the reabsorbate osmolality to luminal osmolality; this mechanism is far more significant than any other parameter change.     

Relationship between streaming potential and compressive stress in bovine intervertebral tissue

The intervertebral disc is formed by the nucleus pulposus (NP) and annulus fibrosus (AF), and intervertebral tissue contains a large amount of negatively charged proteoglycan. When this tissue becomes deformed, a streaming potential is induced by liquid flow with positive ions. The anisotropic property of the AF tissue is caused by the structural anisotropy of the solid phase and the liquid phase flowing into the tissue with the streaming potential. This study investigated the relationship between the streaming potential and applied stress in bovine intervertebral tissue while focusing on the anisotropy and loading location. Column-shaped specimens, 5.5 mm in diameter and 3 mm thick, were prepared from the tissue of the AF, NP and the annulus–nucleus transition region (AN). The loading direction of each specimen was oriented in the spinal axial direction, as well as in the circumferential and radial directions of the spine considering the anisotropic properties of the AF tissue. The streaming potential changed linearly with stress in all specimens. The linear coefficients k_e of the relationship between stress and streaming potential depended on the extracted positions. These coefficients were not affected by the anisotropy of the AF tissue. In addition, these coefficients were lower in AF than in NP specimens. Except in the NP specimen, the k_e values were higher under faster compression rate conditions. In cyclic compression loading the streaming potential changed linearly with compressive stress, regardless of differences in the tissue and load frequency.