Risk factors for osteoarthritis include raised BMI and female gender. Whether these two factors influenced synovial gene expression was investigated using a triangulation and modelling strategy which generated 12 datasets of gene expression in synovial tissue from three knee pathologies with matching BMI groups, obese and overweight, and gender distributions. Intra-operative synovial biopsies were immersed in RNAlater at 4oC before storage at -80oC. Total RNA was extracted using RNAeasy with gDNA removal. Following RT- PCR and quality assessment, cDNA was applied to Affymetrix Clariom D microarray gene chips. Bioinformatics analyses were performed. Linear models were prepared in limma with gender and BMI factors incorporated sequentially for each pathology comparison, generating 12 models of probes differentially expressed at FDR p<0.05 and Bayes number, B>0. Data analysis of differently expressed genes utilized Ingenuity Pathway Analysis and Cytoscape with Cluego and Cytohubba plug-ins.Abstract
Introduction
Methodology
It is increasingly evident that synovium may play a larger role in the aetiology of osteoarthritis. We compared gene expression in whole tissue synovial biopsies from end-stage knee osteoarthritis and knee trauma patients with that of their paired explant cultures to determine how accurately cultured cells represent holistic synovial function. Synovial tissue biopsies were taken from 16 arthroplasty patients and 8 tibial plateau fracture patients with no osteoarthritis. Pairs of whole tissue fragments were either immediately immersed in RNAlater Stabilisation Solution at 4o C before transfer to -80o C storage until RNA extraction; or weighed, minced and cultured at 500mg tissues/5ml media in a humidified incubator at 37oC, 5% CO2. After sub-culturing total RNA was extracted using RNAeasy Plus Mini Kit with gDNA removal. Following RT-PCR and quality assessment, cDNA was applied to Affymetrix Clariom D microarray gene chips. Bioinformatics analyses were performed.Abstract
Introduction
Methodology
Articular cartilage degradation is a defining feature of osteoarthritis. Synovium is a reactive tissue with synovial villae, neoangiogenesis and intimal hyperplasia common to many joint pathologies. The consequences of cartilage debris in osteoarthritis impacting the synovial intima is not well understood. We analysed the immunohistology of synovium from 16 patients with osteoarthritis and 17 patients undergoing knee surgery for non-arthritic pathologies. This data was integrated with imaging and functional scores to correlate synovitis in osteoarthritis. Formalin-fixed paraffin embedded synovial biopsy sections were cut in serial sequence and processed for routine staining (H&E or CD3, CD68, CD20, Vimentin, vWF and PCNA IHC) using standardised Dako monoclonal mouse anti-human antibodies. Digital images scanned at x20 were evaluated for fragments of cartilage and aggregates of inflammatory cells. Clinical data (gender, BMI, KL grade, WOMAC & SF-12 scores) was aligned with histopathological data.Abstract
Introduction
Methodology
Synovitis impacts osteoarthritis symptomatology and progression. The transcription factors controlling synovial gene expression have not been described. This study analyses gene expression in synovium samples from 16 patients with osteoarthritis with 9 undergoing arthroscopic and 8 knee trauma surgery for non-arthritic pathologies. Intra-operative synovial biopsies were immersed in RNAlater at 4oC before storage at -80oC. Total RNA was extracted using RNAeasy. After purification, RT-PCR and quality assessment, cDNA was applied to Affymetrix Clariom D microarray gene chips. Bioinformatics analyses were performed. Linear models were prepared in limma with gender and BMI factors incorporated sequentially for each pathology comparison, generating 12 models of probes differentially expressed at FDR p<0.05 and Bayes number, B>0. Data analysis of differently expressed genes utilized Ingenuity Pathway Analysis and Cytoscape with Cluego and Cytohubba plug-ins.Abstract
Introduction
Methodology
In patients with neural disorders such as cerebral palsy, three-dimensional marker-based motion analysis has evolved to become a well standardized procedure with a large impact on the clinical decision-making process. On the other hand, in knee arthroplasty research, motion analysis has been little used as a standard tool for objective evaluation of knee joint function. Furthermore, in the available literature, applied methodologies are diverse, resulting in inconsistent findings [1]. Therefore we developed and evaluated a new motion analysis framework to enable standardized quantitative assessment of knee joint function. The proposed framework integrates a custom-defined motion analysis protocol with associated reference database and a standardized post-processing step including statistical analysis. Kinematics are collected using a custom-made marker set defined by merging two existing protocols and combine them with a knee alignment device. Following a standing trial, a star-arc hip motion pattern and a set of knee flexion/extension cycles allowing functional, subject-specific calibration of the underlying kinematic model, marker trajectories are acquired for three trials of a set of twelve motor tasks: walking, walking with crossover turn, walking with sidestep turn, stair ascent, stair descent, stair descent with crossover turn, stair descent with sidestep turn, trunk rotations, chair rise, mild squat, deep squat and lunge. This specific set of motor tasks was selected to cover as much as possible common daily life activities. Furthermore, some of these induce greater motion at the knee joint, thus improving the measurement-to-error ratio. Kinetics are acquired by integrating two forceplates in the walkway. Bilateral muscle activity of 8 major muscles is monitored with a 16 channel wireless electromyography (EMG) system. Finally, custom-built software with an associated graphical user interface was created for automated and flexible analysis of gait lab data, including repeatability analysis, analysis of specific kinematic, kinetic and spatiotemporal parameters and statistical comparisons.INTRODUCTION
MATERIALS AND METHODS
After total knee arthroplasty (TKA) with a PCL-retaining implant the location of the tibiofemoral contact point should be restored in order to obtain normal kinematics. The difficulty during surgery is to control this location since the position of the femur on the tibia cannot easily be measured from the back of the joint. Therefore, we developed a simple “spacer technique” to check the contact point indirectly in 90° flexion after all bone cuts are made by measuring the step-off between the distal cut of the femur and the anterior edge of the tibia with a spacer in place. The goal of this experiment was to investigate whether this new PCL balancing approach with the spacer technique created the correct contact point location. Nine fresh-frozen full leg cadaver specimens were used. After native testing, prototype components of a new PCL-retaining implant were implanted using navigation and a bone-referenced technique. After finishing the bone cuts of tibia and femur, the spacer was inserted in flexion and positioned on the anterior edge of the bony surface to measure the step-off. If necessary, an extra cut was made to balance the PCL. The specimen was mounted on the knee kinematics rig and a squat with constant vertical ankle force (130N) and constant medial and lateral hamstrings forces (50N) was performed between 30° and 130° of knee flexion. The trajectories of the reflective tibial and femoral markers were continuously recorded using six infrared cameras. The projections of the femoral condylar centers on the horizontal plane of the tibia were calculated and compared.Introduction
Methods
Proper positioning of the components of a knee prosthesis for obtaining post-operative knee joint alignment is vital to obtain good and long term performance of a knee replacement. Although the reasons for failure of knee arthroplasty have not been studied in depth, the few studies that have been published claim that as much as 25% of knee replacement failures are related to malpositioning or malalignment [x]. The use of patient-matched cutting blocks is a recent development in orthopaedics. In contrast to the standard cutting blocks, they are designed to fit the individual anatomy based on 3D medical images. Thus, landmarks and reference axes can be identified with higher accuracy and precision. Moreover, stable positioning of the blocks with respect to the defined axes is easier to achieve. Both may contribute to better alignment of the components. The objective of this study was to check the accuracy of femoral component orientation in a cadaver study using specimen-matched cutting blocks in six specimens; first for a bi-compartmental replacement, and then for a tri-compartmental replacement in the same specimen. Frames with infrared reflective spherical markers were fixed to six cadaveric femurs and helical CT scans were made. A bone surface reconstruction was created and the relevant landmarks for describing alignment were marked using 3D visualisation software (Mimics). The centres of the spherical markers were also determined. Based on the geometry of the articular surface and the position of the landmarks, custom-made cutting blocks were designed. One cutting block was prepared to guide implantation of a bi-compartmental device and another one to guide implantation of the femoral component of a total knee replacement. The knee was opened and the custom-made cutting block for the bi-compartmental implant was seated onto the surface. The block was used to make the anterior cut, after which it was removed and replaced with the conventional cutting block using the same pinning holes to ensure the same axial rotational alignment. The other cuts were made using the conventional cutting block and the bi-compartmental femoral component was implanted. Afterwards, a similar procedure was used to make the extra cuts for the total knee component. The position of the components with respect to the reflective markers was measured by locating three reference points and “painting” the articular surface with a wand with reflective markers. The position of all marker spheres was continuously recorded with four infrared cameras and Nexus software.Purpose
Materials and Methods
A comparative kinematic study was carried out on six cadaver limbs, comparing tibiofemoral kinematics in five different conditions: unloaded, under a constant 130 N ankle load with a variable quadriceps load, with and without a constant 50 N medial and lateral hamstrings load. Kinematics were described as translation of the projected centers of the medial (MFT) and lateral femoral condyles (LFT) in the horizontal plane of the tibia, and tibial axial rotation (TR) as a function of flexion angle. In passive conditions, the tibia rotated internally with increasing flexion, to an average of −16° (range −12/−20°, SD 3.0°). Between 0 – 40° flexion, the medial condyle translated forwards 4 mm (range 0.8/5.5 mm, SD 2.5 mm), followed by a gradual posterior translation, totaling −9 mm (range −5.8/−18.5 mm, SD 4.9 mm) between 40° – 140° flexion. The lateral femoral condyle translated posteriorly with increasing flexion completing −25 mm (range −22.6 – −28.2 mm, SD 2.5 mm). Dynamic, loaded measurements were carried out in a knee rig. Under a fixed ankle load of 130 N and variable quadriceps loading, tibial rotation was inverted, mean TR 4.7° (range −3.3°/11.8° SD 5.4°), MFT −0.5 mm (range = −4.3/2.4 mm, SD = 2.4 mm), LFT 3.3 mm (range = −3.6/10.6 mm, SD = 5.1 mm). As compared to the passive condition, all these excursions were significantly different: p=0.015, p=0.013, and p=0.011 for TR, MFT and LFT respectively. Adding medial and lateral hamstrings force of 50N each, reduced TR, MFT and LFT significantly as compared to the passive condition. In general, loading the knee with hamstrings and quadriceps reduces rotation and translation as compared to the passive condition. Lateral hamstring action is more influential on knee kinematics than medial hamstrings action.
