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.

Background

We reproduced a frequently cited study performed at our University Hospital that was published in the British Medical Journal in 1981 assessing the extent of “snow and ice” fractures during the winter period.

Osteoarthritis (OA)

is the most common arthritic condition. OA causes joint pain, loss of mobility and significantly affects the quality of life for the affected individual. The major burden to patients with arthritis is pain. However, often radiological joint destruction and the extent of pain do not correlate. This causes a dilemma for clinicians in advising timing for joint replacement surgery. In arthritis, concentrations of the neurotransmitter, glutamate is increased within the synovial fluid activating both peripheral pain mechanisms and pathological processes (1). Other pathological/pain related metabolites are also released into synovial fluid, which provides a real time snap shot of the joint pathology. We have tested the hypothesis that ‘The increased levels of pain and disease-related metabolites within human synovial fluids from arthritic joints can be detected and quantified ex vivo using high resolution 1H-NMR.’

Fractures of the shaft of the humerus are often treated conservatively in a hanging cast or a humeral brace. The conservative management of this fracture is often prolonged and quite uncomfortable for the patient. Some of the patients will need an operative fixation after a trial of conservative management.

We retrospectively looked at 72 consecutive patients with fractures of the shaft of the humerus that presented in our institution over a period of two years. The fracture pattern, treatment modality time to union and the number that needed operative fixation following a trial of conservative treatment was analysed. Of the 72 patients 4 were lost to follow-up. 45 patients had a 1.2.B or 1.2.C type of fracture and 23 had a 1.2.A type of fracture. 29 (41%) were successfully treated conservatively, 11 (16%) patients were operated as the primary procedure and 15 (22%) patients were operated due to delayed or non union. 13 (19%) patients were operated within 4 weeks of the fracture as their alignment was not acceptable on their weekly follow-up.

The average time to union in the patients treated conservatively was 22 weeks, while that of the patients treated primarily by open reduction and plating was 14 weeks (p-value<0.05). Patients who needed operation after initial conservative management required prolonged period of rehabilitation and union time was 32.2 weeks. At the time of fracture union 72% of the patients who had been treated conservatively had joint stiffness requiring physiotherapy, while only 18% of those who had an open reduction and internal fixation had stiffness and required physiotherapy. (p-value < 0.05).

In conclusion careful consideration should be given before it is decided to treat this fracture conservatively especially in the case of 1.2.A fracture pattern.

Background: Acute traumatic rupture of the Tendo-Achilles is a common problem. Review of the published literature shows a conflict between surgical and non-surgical management. Primarily, surgical treatment has repeatedly been shown to have a lower re-rupture rate compared with conservative treatment. However, it is associated with a higher rate of other complications, principally infective soft tissue complications. Debate concerning the method and duration of conservative treatment regimens also remains.

Methods: We aimed to assess the effect of knee position on the gap in acute tendo-Achilles ruptures. Our null hypothesis was there is no difference in tendon gap with variation of knee position. Patients with a history and examination consistent with an acute rupture were collected prospectively. They were imaged by the same consultant musculoskeletal consultant radiologist using ultrasound. Tendon gap was measured with the foot in neutral and full tolerable equinus with the knee both flexed to 90° and fully extended. A pilot study of 10 patients was used to provide data for a power calculation and consequently a total of 25 patients were recruited.

Results: All patients had their clinical diagnosis confirmed and tendon gap measured in each of the 4 positions. A paired t-test was used to compare mean differences between each position. This confirmed there was no significant difference in the tendon gap with the knee flexed or extended providing the foot is in full equinus. This may have considerable implications on the rationale behind conservative treatment and splinting of acute tendo-Achilles ruptures.

We aimed to determine the reliability, accuracy and consequently the clinical role of digital templating in the pre-operative work up for total knee arthroplasty patients.

With the increasing use of digital radiology images, analogue templating may soon be defunct. Digital templating is a more recent development and its role is yet to be determined.

Ten pre-operative digital radiographs were templated by four independent observers. Inter-observer and intra-observer reliability was assessed using the kappa measure of concordance. Subsequently, 40 consecutive total knee arthroplasty patients underwent pre-operative digital templating. This was a blinded process by a consultant surgeon not involved with the operation. Each patient underwent TKR using the PFC Sigma System sized intra-operatively, without the operating surgeon having knowledge of the pre-operative templating result. Comparison was made between the pre-operative digital templates and the blinded intra-operative sizing.

For both the femoral and tibial templating there was good to very good inter- and intra-observer agreement. For the femoral component the templating was correct in 47.5% (± 1 size difference 97.5%). The tibial templating was correct in 55% (± 1 size difference 100%).

The inter- and intra-observer reliability of digital templating process has been shown to be acceptable but the correlation between digital templating and the actual size implanted is poor. Our series shows a similar accuracy to the published data on analogue templating for the same implant. Like analogue templating, its clinical role remains uncertain and its poor correlation to the actual implant sizes limits its usefulness.

We aimed to determine the reliability, accuracy and the clinical role of digital templating in the pre-operative work-up for total knee replacement.

Initially a sample of ten pre-operative digital radiographs were templated by four independent observers to determine the inter- and intra-observer reliability of the process. Digital templating was then performed on the radiographs of 40 consecutive patients undergoing total knee replacement by a consultant surgeon not involved with the operation, who was blinded to the size of the implant inserted. The Press Fit Condylar Sigma Knee system was used in all the patients. The size of the implant as judged by templating was then compared to that of the size used.

Good inter- and intra-observer agreement was demonstrated for both femoral and tibial templating. However, the correct size of the implant was predicted in only 48% of the femoral and 55% of the tibial components. Albeit reproducible, digital templating does not currently predict the correct size of component often enough to be of clinical benefit.

Introduction: Monoclonal antibodies (mAbs) recognizing linear sulphation motifs in keratan sulphate (KS) were first developed in the early 1980’s. Over the years, ELISAs using 5-D-4 or other related anti-KS mAbs have been used in many studies monitoring increased cartilage aggrecan degradation with the onset of degenerative joint diseases. However, whilst these studies have in general been useful for monitoring some aspects of disease progression (usually in parallel with other biomarker assays), many longitudinal studies have shown efficacy in only the transient (early, mid or late) stages of the degenerative joint disease process. During the onset of degenerative joint disease, the pathological tissue attempts to repair/regenerate the cartilage, the chondrocytes thus synthesizing cartilage aggrecan with KS substitution [and chondroitin sulphate (CS) isomer composition] that is more like that found in developing or immature cartilage. This immature cartilage aggrecan contains much less KS substitution with shorter chain size and less linear sulphation motifs. Thus, during the different stages of degenerative joint disease progression one would expect to find variable changes in different linear sulphation epitopes present in the serum or synovial fluids. The aim of this study was to investigate the use of several monoclonal antibodies that recognise different sulphation epitopes [high sulphation (5-D-4), low sulphation (1-B-4) and KS-stubs (BKS-1)] to see if patterns of their expression could be used to distinguish different stages of degenerative joint disease. We have also developed ELISAs using mAbs recognising the KS-proteoglycans, keratocan (Ker 1) and lumican (Lum 1) for their quantification as potential biomarkers of osteoarthritis.

Methods: Competitive ELISAs were developed using monoclonal antibodies (mAbs) 5-D-4, 1B4, BKS-1, Ker-1 and Lum-1. Bovine corneal KS-proteoglycans pre-treated with keratanase were used as both the coating antigen and “standard” antigen on the same ELISA plate. Blood, synovial fluid and cartilage samples (surgical waste) obtained from patients undergoing arthroplasty with different Kellgren & Lawrence grades were analysed.

Results and Discussion: 5-D-4 and BKS-1 showed similar inhibition curves and relative 50% inhibition points. However, the curve obtained with 1B4 indicated lower relative expression of 1B4 epitope. Analysis of serum and synovial fluid sample with 5-D-4 mAb showed the presence of the epitope in both samples, but there was significantly less KS in serum than in the synovial fluid. Our results show that competitive ELISA for quantification of several different KS sulphation or “stub” epitopes and two KS-proteoglycans can all be quantified and compared using the same experimental conditions. These studies are ongoing as part of an Arthritis Research Campaign (UK) funded study. In addition the data indicates that keratocan and lumican are also increased in their expression with the progression of disease. Future studies will be performed in an attempt to quantify increased keratocan and lumican expression as potential biomarkers of degenerative joint disease.

Introduction: Kashin-Beck disease (KBD) is an endemic osteoarthropathy with pathological changes occurring in growth plate and articular cartilage in humans. It manifests as cartilage degeneration and necrosis. It is postulated that KBD is due to fungal mycotoxins infiltrating the diet and a regional selenium deficiency in the environment providing food sources in a broad belt across China. Previous work has established an in vitro system in which chondrocytes are cultured and an ex vivo cartilage graft is produced. Subjecting these chondrocytes to either selenium (SEL), Nivalenol (NIV) or in combination during the growth of the graft was found to alter the morphology of the cartilage graft. In addition, the quantity of the large aggregating proteoglycan, was significantly reduced in a dose dependent manner in the presence of Nivalenol. This study aimed to examine the composition of aggrecan from grafts grown in the presence of NIV or SEL alone, or in combination to better understand cellular and molecular mechanisms underlying the pathogenesis of KBD.

Methods: Chondrocytes (from 7 day old bovine cartilage) were seeded at high density in MilliCell filter inserts (12mm diameter; Millipore, MA). Cultures were maintained for 4 weeks in DMEM supplemented with 20% heat–inactivated FBS, ascorbate (100μg/ml) and TGFß2 (5ng/ml) or additionally supplemented with either SEL , NIV or both at concentrations of 0.01, 0.05 and 0.1μg/ml. Media was refreshed thrice weekly and later analysed. At 4 weeks the cartilage grafts were harvested, weighed and extracted in 4M guanidium chloride (with an inhibitor cocktail) for biochemical analysis of matrix molecules. Residues were papain digested. Glycosaminoglycan concentration was determined using the DMMB assay in all media samples, guanidine extracts and papain digests. Aggrecan and GAG composition was determined using Western blotting with a panel of antibodies recognising chondroitin sulphate (CS), keratan sulphate (KS) and protein core epitopes present in aggrecan.

Results: The total GAG synthesised in a 4week period was substantially reduced in chondrocytes cultured in the presence of NIV at 0.05 and 0.1μg/ml and to a lesser extent in those cultures exposed to the highest dose of SEL. However, the amount of GAG released into the media remained fairly constant within the treatment groups, but a marked reduction was apparent in the guanidine extracts of the cartilage grafts. Western blot analysis with a series of antibodies on guanidine extracted aggrecan showed no substantial changes in the core protein molecular weights however analysis demonstrated that KS was reduced in NIV treated cultures. Results also indicated that NIV treated cultures appeared to contain less CS substitutions on the aggrecan core protein.

Discussion: The GAG concentration data indicates that there is an inability of the GAG to remain within the cartilage grafts extracellular matrix. when treated with NIV. Western blot analysis indicates minor changes in the composition of the aggrecan in relation to protein core length and CS/KS side chain substitutions or length. Further work will investigate the proportion of aggrecan able to form high molecular weight aggregates, the metabolism of link protein and hyaluronan.