Introduction

The prevalence of symptomatic osteoarthritis (OA) in the knee is 11–11% compared to 3.4–4.4% in the ankle. In addition to this, 70% of ankle arthritis is post-traumatic while the vast majority of knee arthritis is primary OA. Several reports have previously implicated biochemical differences in extracellular matrix composition between these joint cartilages; however, it is unknown whether there is an inherent difference in their transcriptome and how this might affect their respective functionality under load, inflammatory environment etc. Therefore, we have analysed the transcriptome of ankle and knee cartilage chondrocytes to determine whether this could account for the lower prevalence and altered aetiology of ankle OA.

The lifetime prevalence of symptomatic osteoarthritis at the knee is 50% osteoarthritis of the ankle occurs in only 1% of the population. This variation in prevalence has been hypothesised to result from the differential responsiveness of the joint cartilages to catabolic stimuli.

Human cartilage explants were taken from the talar domes (n=12) and the femoral condyles (n=7) following surgical amputation. Explants were cultured in the presence of either a combination of high concentration cytokines (TNFα, OSM, IL-1α) to resemble a post traumatic environment or low concentration cytokines to resemble a chronic osteoarthritic joint. Cartilage breakdown was measured by the percentage loss of Sulphated glycosaminoglycan (sGAG) from the explant to the media during culture. Expression levels of the pro-inflammatory molecules nitric oxide and prostaglandin E₂ were also measured.

Significantly more sGAG was lost from knee cartilage exposed to TNFα (22.2% vs 13.2%, P=0.01) and TNFα in combination with IL-1α (27.5% vs 16.0%, P=0.02) compared to the ankle; low cytokine concentrations did not affect sGAG release. Significantly more PGE₂ was produced by knee cartilage compared to ankle cartilage however no significant difference in nitrite production was noted.

Cartilage from the knee and ankle has a divergent response to stimulation by pro-inflammatory cytokines, with high concentrations of TNFα alone, or in combination with IL-1α amplifying cartilage degeneration. This differential response may account for the high prevalence of knee arthritis compared to ankle OA and provide a future pharmacological target to treat post traumatic arthritis of the knee.

One reason why NICE (National Institute for Clinical Excellence) does not support operations by the NHS to heal hyaline cartilage lesions using a patients own cells is because there is no clear evidence to show that these operations are beneficial and cost-effective in the long term. Specifically, NICE identified a deficiency of high quality cartilage being produced in repaired joints. The presence of high quality cartilage is linked to long-lasting and functional repair of cartilage. The benchmark for quality, NICE stipulate, is repair cartilage that is stiff and strong and looks similar to the normal tissue surrounding it, i.e. mature hyaline articular cartilage.

Biopsy material from autologous cartilage implantation surgical procedures has the appearance of immature articular cartilage and is frequently a mixture of hyaline and fibrocartilage. Osteoarthritic cartilage, in its early stages, also exhibits characteristics of immature articular cartilage in that it expresses proteins found in embryonic and foetal developmental stages, and is highly cellular as evidenced through the presence of chondrocyte clusters. Therefore, an ability to modulate the phenotype and the structure of the extracellular matrix of articular cartilage could positively affect the course of repair and regeneration of articular cartilage lesions. In order to do this, the biochemical stimuli that induce the transition of an essentially unstructured amorphous cartilage mass (immature articular cartilage) to one that is highly structured and ordered, and biomechanically adapted to its particular function (mature articular cartilage) has to be identified.

We show for the first time, that fibroblast growth factor-2 and transforming growth factor beta-1 induce precocious maturation of immature articular cartilage. Our data demonstrates that it is possible to significantly enhance maturation of cartilage tissue using growth factor stimulation; consequently this may have applications in transplantation therapy, or through phenotypic modulation of osteoarthritic chondrocytes in diseased cartilage in order to stimulate growth and maturation of repair tissue.

Patient specific knee modelling has the potential to help understand the development of the mechanically induced degenerative disease, Osteoarthritis. A full joint contact model of the knee involves modelling the bones, ligaments, articular cartilage (AC) and meniscus, as well as, the kinematics and geometry of real joints. These finite element models will inevitably require great computational resource to run and it is desirable to find resource effective material model formulations which can accurately describe the mechanical behaviour of the soft tissues. Biphasic models (BIMs) have long been established as an effective formulation for modelling AC. However, the swelling behaviour caused by changes in the ionic phase is a major recovery mechanism and is neglected in the BIMs. It is therefore believed that BIMs alone are insufficient to fully describe the mechanical behaviour of AC. Instead, a thermal analogy method which is generically a BIM that includes the swelling behaviour has been thought to be suitable and has been validated against literature data using material parameters optimized to match the numerical and experimental results. To ensure the model is suitable for patient specific modelling where it will have the ability to reflect the individual AC material properties of the patients in the mechanical behaviour it predicts, two experiments have been planned and are currently being carried out using bovine AC. The first experiment is to investigate the diffusivity of the tissue in solutions of different molarity by measuring the change in tissue weight over time. Eleven explants are taken from the same bovine articular joint using a 6mm biopsy punch and are left in 10mM of PBS overnight to ensure ionic equilibrium has been reached before experiments are carried out. The explants are then placed in PBS solutions of molarities ranging from 0mM to 10mM and weighed at regular time intervals. In the final stage, the explants are then lyophilized and weighed for determining the volume of water in the tissues. Using Archimedes principle, the change in porosity of the tissue is found. A preliminary study has shown that explants submerged in a solution of 5mM has an approximately 4% change in weight after the first 24h and a further 1.73% change in the following 24h. Control specimens left in a solution of 10mM had a 0% change in weight. The second experiment is to carry out mechanical loading on the AC specimens while submerged in a solution of different ion concentrations. Experiments with various loading conditions are being investigated to explore their efficacy for validation. Preliminary compression tests have been carried out where steps of 1% strain was applied, giving a total of 10% strain. Between each step, strain was held constant until full relaxation has been achieved. The reaction force measured from the second experiment in conjunction with data collected from the first experiment will be compared to results predicted in the numerical model. This will allow the determination of whether thermal analogy is adequate or whether more complex triphasic models need to be considered. Furthermore, the development of these experimental methods will contribute to the validation of other AC material models in the future.

Introduction

Low back pain is a major public health problem in our society. Degeneration of intervertebral disc (IVD) appears to be the leading cause of chronic low-back pain [1]. Mechanical stimulations including compressive and tensional forces are directly implicated in IVD degeneration. Several studies have implicated the cytoskeleton in mechanotransduction [2, 3], which is important for communication and transport between the cells and extracellular matrix (ECM). However, the potential roles of the cytoskeletal elements in the mechanotransduction pathways in IVD are largely unknown.