Lectin-binding in normal and osteoarthrotic articular cartilage from STR/1N-mouse knee joints

Fluorescein-isothiocyanate (FITC) labeled lectins were used to study the distribution pattern of specific binding-sites in histological sections of normal and osteoarthrotic articular cartilage from the mouse knee joint. Male inbred mice of the STR/1N-strain develop spontaneous arthrotic articular cartilage lesions on the medial condyle of tibia and femur. The varus-deformity of the knee joint leads to a recurrent medial patellar luxation with osteoarthrotic defects on the medial part of the facies patellaris femoris. It was demonstrated that the lectin staining pattern of osteoarthrotic articular cartilage, especially on the facies patellaris femoris, was different from that of normal articular cartilage. The differences in lectin staining corresponded to those observed between normal and fibrillated articular cartilage from human patellae. The normal articular cartilage of the mouse knee joint possessed lectin binding-sites for Concanavalin A (ConA) and wheat germ agglutinin (WGA), but not for Ulex europaeus agglutinin (UEA), soy bean agglutinin (SBA) and peanut agglutinin (PNA). In addition to the completely changed distribution pattern of ConA and WGA in osteoarthrotic cartilage, SBA, PNA and UEA developed distinct staining patterns particular to the fibrillated areas of arthrotic cartilage. The increased lectin-binding to arthrotic articular cartilage may be due to unmasking of sugars in the course of bondage breakdown in fibrillated cartilage or the production of pathological glycoproteins. It is evident that lectins can demonstrate minute differences between normal and arthrotic cartilage and it is concluded, therefore, that lectins are sensitive and specific tools for the study of degenerative joint diseases.     

Composition of the medial and posterior articular nerves of the mouse knee joint

The number and the distribution of fiber size in the medial (MAN) and posterior (PAN) articular nerves of the mouse knee joint were studied by electron microscopy. The MAN contained 75 +/- 28 nerve fibers consisting of 63 +/- 24 unmyelinated and 12 +/- 6 myelinated fibers. The PAN was composed of 195 +/- 50 nerve fibers, namely 129 +/- 28 unmyelinated and 66 +/- 24 myelinated fibers. A skewed unimodal distribution of the unmyelinated nerve fiber diameters was seen in both nerves ranging from 0.1 to 1.2 microm with a maximum between 0.3 and 0.6 microm. The myelinated nerve fibers in the MAN ranged from 1 to 8 microm with a peak between 2 and 5 microm. In the PAN, their diameters ranged from 1 to 12 microm with a clearly visible peak at 4-5 microm and a plateau at 8-9 microm that may represent a second maximum. These data show that the knee joint innervation of the mouse is comparable to those of the cat and rat concerning the types of nerve fibers and the composition of the two nerves. However, in relation to the much smaller area of tissue to be innervated the total number of primary afferents is considerable smaller in the mouse.     

Fibroblast-like synovial cells from normal and inflamed knee joints differently affect the expression of pain-related receptors in sensory neurones: a co-culture study

Innervation of the joint with thinly myelinated and unmyelinated sensory nerve fibres is crucial for the occurrence of joint pain. During inflammation in the joint, sensory fibres show changes in the expression of receptors that are important for the activation and sensitization of the neurones and the generation of joint pain. We recently reported that both neurokinin 1 receptors and bradykinin 2 receptors are upregulated in dorsal root ganglion (DRG) neurones (the cell bodies of sensory fibres) in the course of acute and chronic antigen-induced arthritis in the rat. In this study, we begin to address mechanisms of the interaction between fibroblast-like synovial (FLS) cells and sensory neurones by establishing a co-culture system of FLS cells and DRG neurones. The proportion of DRG neurones expressing neurokinin 1 receptor-like immunoreactivity was not altered in the co-culture with FLS cells from normal joints but was significantly upregulated using FLS cells from knee joints of rats with antigen-induced arthritis. The proportion of DRG neurones expressing bradykinin 2 receptors was slightly upregulated in the presence of FLS cells from normal joints but upregulation was more pronounced in DRG neurones co-cultured with FLS cells from acutely inflamed joints. In addition, the expression of the transient receptor potential V1 (TRPV1) receptor, which is involved in inflammation-evoked thermal hyperalgesia, was mainly upregulated by co-culturing DRG neurones with FLS cells from chronically inflamed joints. Upregulation of neurokinin 1 receptors but not of bradykinin 2 and TRPV1 receptors was also observed when only the supernatant of FLS cells from acutely inflamed joint was added to DRG neurones. Addition of indomethacin to co-cultures inhibited the effect of FLS cells from acutely inflamed joints on neurokinin 1 receptor expression, suggesting an important role for prostaglandins. Collectively, these data show that FLS cells are able to induce an upregulation of pain-related receptors in sensory neurones and, thus, they could contribute to the generation of joint pain. Importantly, the influence of FLS cells on DRG neurones is dependent on their state of activity, and soluble factors as well as direct cellular contacts are crucial for their interaction with neurones.     

The optic nerve in the cat. II. Qualitative aspects of maturation in the adult cat

54 healthy cats were studied. In almost all cases, two types of degenerative signs could be seen: in the primary optic pathways, but also in the posterior funiculi and the spinal-cerebellar tracts, lesions of wide-diameter axons seen by their initial demyelination, accompanied by intense neuroglial reactions, and, on the other hand, a Wallerian degeneration of smaller axons situated in the axial part of the nerve. The percentage of these abnormalities must still be evaluated. Their eventual consequences and nature are discussed.     

Quantitative studies on the regeneration of myelinated nerve fibers. I. Variations in the number and the size of myelinated fibers in the nerves of normal rats]

The number and size of myelinated nerve fibers have been determined at standard levels in the nerve to medial head of right and left gastrocnemius muscles of 24 normal rats (11 males and 13 females). The mean values of all results were comparable on right and left sides. Thus, 271 +/- 5 myelinated nerve fibers were found in the right nerve and 272 +/- 4 in the left; their mean diameter were respectively 8.1 +/- 0.1 and 8.0 +/- 0.1 micron. There were 60.1% of large nerve fibers on the right side and 59,9% on the left, their mean diameters being 10.5 and 10.6 micron. Some variations occured in all these values, depending of the weight and sex of the animals. Nevertheless, the differences between both sides of a same rat were negligible and the histograms of both nerves could be superposed. Accordingly, in the operated animals, the contralateral nerve may be used as control.     

Comparison of two methods for calculating the frictional properties of articular cartilage using a simple pendulum and intact mouse knee joints

In attempts to better understand the etiology of osteoarthritis, a debilitating joint disease that results in the degeneration of articular cartilage (AC) in synovial joints, researchers have focused on joint tribology, the study of joint friction, lubrication, and wear. Several different approaches have been used to investigate the frictional properties of articular cartilage. In this study, we examined two analysis methods for calculating the coefficient of friction (μ) using a simple pendulum system and BL6 murine knee joints (n=10) as the fulcrum. A Stanton linear decay model (Lin μ) and an exponential model that accounts for viscous damping (Exp μ) were fit to the decaying pendulum oscillations. Root mean square error (RMSE), asymptotic standard error (ASE), and coefficient of variation (CV) were calculated to evaluate the fit and measurement precision of each model. This investigation demonstrated that while Lin μ was more repeatable, based on CV (5.0% for Lin μ; 18% for Exp μ), Exp μ provided a better fitting model, based on RMSE (0.165° for Exp μ; 0.391° for Lin μ) and ASE (0.033 for Exp μ; 0.185 for Lin μ), and had a significantly lower coefficient of friction value (0.022±0.007 for Exp μ; 0.042±0.016 for Lin μ) (p=0.001). This study details the use of a simple pendulum for examining cartilage properties in situ that will have applications investigating cartilage mechanics in a variety of species. The Exp μ model provided a more accurate fit to the experimental data for predicting the frictional properties of intact joints in pendulum systems.     

Role of hypoxia-inducible factor 1alpha in the integrity of articular cartilage in murine knee joints

Introduction  

Chondrocytes have to withstand considerable hypoxic conditions within the avascular articular cartilage. The present study investigated the effects of inhibiting or stabilizing hypoxia-inducible factor (HIF)-1α by 2-methoxyestradiol or dimethyloxaloylglycine on the progression of osteoarthritis in murine knee joints.     

Single cilia in the articular cartilage of the cat.

Single cilia (SC) in the articular cartilage of the cat are described with regard to structure, frequency and intracellular location. Comparison is made with their occurrence in the cartilage of other species or in other tissues; special consideration is given to their developmental stages. In the 1-year-old cat examined, about 20% of the cells showed SC with a predominantly intracellular course and some of them did not reach the extracellular space at all. Cross-sections of the proximal region of SC reveal the familiary 9+0 pattern; towards the distal end, the number of doublets and the diameter of the shaft are decreasing. It is concluded that most of the SC encountered are undergoing involution.     

Topographical variation of the elastic properties of articular cartilage in the canine knee

Equilibrium response of articular cartilage to indentation loading is controlled by the thickness (h) and elastic properties (shear modulus, mu, and Poisson's ratio, nu) of the tissue. In this study, we characterized topographical variation of Poisson's ratio of the articular cartilage in the canine knee joint (N=6). Poisson's ratio was measured using a microscopic technique. In this technique, the shape change of the cartilage disk was visualized while the cartilage was immersed in physiological solution and compressed in unconfined geometry. After a constant 5% axial strain, the lateral strain was measured during stress relaxation. At equilibrium, the lateral-to-axial strain ratio indicates the Poisson's ratio of the tissue. Indentation (equilibrium) data from our prior study (Arokoski et al., 1994. International Journal of Sports Medicine 15, 254-260) was re-analyzed using the Poisson's ratio results at the test site to derive values for shear and aggregate moduli. The lowest Poisson's ratio (0.070+/-0.016) located at the patellar surface of femur (FPI) and the highest (0.236+/-0.026) at the medial tibial plateau (TMI). The stiffest cartilage was found at the patellar groove of femur (micro=0.964+/-0.189MPa, H(a)=2.084+/-0. 409MPa) and the softest at the tibial plateaus (micro=0.385+/-0. 062MPa, H(a)=1.113+/-0.141MPa). Comparison of the mechanical results and the biochemical composition of the tissue (Jurvelin et al., 1988. Engineering in Medicine 17, 157-162) at the matched sites of the canine knee joint indicated a negative correlation between the Poisson's ratio and collagen-to-PG content ratio. This is in harmony with our previous findings which suggested that, in unconfined compression, the degree of lateral expansion in different tissue zones is related to collagen-to-PG ratio of the zone.     

Constituents and pH changes in protein rich hyaluronan solution affect the biotribological properties of artificial articular joints.

The relationship between the coefficient of friction and pH value or protein constituents of lubricating fluid, together with viscosity, were studied within a bearing surface model for artificial joint, ultra-high molecular weight polyethylene (UHMWPE) against stainless steel (SUS), using a mechanical spectrometer. Four lubricants were tested in this study: sodium hyaluronate (HA), HA with albumin, HA with gamma-globulin, and HA with (L)alpha-dipalmitoyl phosphatidylcholine ((L)alpha-DPPC). The coefficient of friction between UHMWPE and SUS in HA with albumin or HA with gamma-globulin varied from 0.035 to 0.070 depending on angular velocity and pH. The coefficient of friction in HA or HA with (L)alpha-DPPC varied from 0.023 to 0.045 depending on angular velocity and pH. The variation in pH for HA with albumin had a large effect on the coefficient of friction at low range of angular velocity with viscosity independence. The variation in pH for HA with gamma-globulin had a large effect on the coefficient of friction with viscosity dependence at high angular velocity. The addition of (L)alpha-DPPC showed a small effect on the coefficient of friction at low angular velocity. This study confirms that the presence of albumin in the lubricant promotes pH dependence and viscosity independence of the tribological properties at low speed while the presence of globulin promotes pH and viscosity independence at low speed and promotes pH and viscosity dependence at high speed in the lubrication of UHMWPE against SUS. This study supports the clinical hypothesis that the effect of constituents and pH changes in periprosthetic fluid for the lubrication is a clue toward resolving many complications after total joint replacement.