Fast and accurate identification of pathogens causing periprosthetic joint infections (PJI) is essential to initiate effective antimicrobial treatment. Culture-based approaches frequently yield false negative results, despite clear signs of infection. This may be due to the use of general growth media, which do not mimic the conditions at site of infection. Possible alternative approaches include DNA-based techniques, the use of In this study, 120 synovial fluid samples were included, aspirated from patients with clinical signs of PJI. For these samples microbiology data (obtained in the clinical microbiology lab using standard procedures) and next generation sequencing (NGS) data, were available. The samples were incubated in the SSF medium at different oxygen levels (21% O2, 3% O2 and 0% O2) for 10 days. Every 24h, the presence of growth was checked. From positive samples, cultures were purified on Columbia blood agar and identified using MALDI-TOF. In parallel, heat produced by metabolically active microorganisms present in the samples was measured using ITC (calScreener, Symcel), (96h at 37°C, in SSF, BHI and thioglycolate). From the resulting thermograms the ‘time to activity’ could be derived. The accuracy and time to detection were compared between the different detection methods.Aim
Methods
There is growing evidence that bacteria encountered in periprosthetic joint infections (PJI) form surface-attached biofilms on prostheses, as well as biofilm aggregates embedded in synovial fluid and tissues. However, models allowing the investigation of these biofilms and the assessment of their antimicrobial susceptibility in physiologically relevant conditions are currently lacking. To address this, we developed a synthetic synovial fluid (SSF) model and we validated this model in terms of growth, aggregate formation and antimicrobial susceptibility testing, using multiple PJI isolates. 17 PJI isolates were included, belonging to Aim
Methods
The aetiologies of common degenerative spine, hip, and knee pathologies are still not completely understood. Mechanical theories have suggested that those diseases are related to sagittal pelvic morphology and spinopelvic-femoral dynamics. The link between the most widely used parameter for sagittal pelvic morphology, pelvic incidence (PI), and the onset of degenerative lumbar, hip, and knee pathologies has not been studied in a large-scale setting. A total of 421 patients from the Cohort Hip and Cohort Knee (CHECK) database, a population-based observational cohort, with hip and knee complaints < 6 months, aged between 45 and 65 years old, and with lateral lumbar, hip, and knee radiographs available, were included. Sagittal spinopelvic parameters and pathologies (spondylolisthesis and degenerative disc disease (DDD)) were measured at eight-year follow-up and characteristics of hip and knee osteoarthritis (OA) at baseline and eight-year follow-up. Epidemiology of the degenerative disorders and clinical outcome scores (hip and knee pain and Western Ontario and McMaster Universities Osteoarthritis Index) were compared between low PI (< 50°), normal PI (50° to 60°), and high PI (> 60°) using generalized estimating equations.Aims
Methods
Cellular therapies play an important role in tendon tissue engineering with tenocytes being described as the most prominent cell population if available in large numbers. However, in vitro expansion of tenocytes in standard culture leads to phenotypic drift and cellular senescence. Recent work suggests that maintenance of tenogenic phenotype in vitro can be achieved by recapitulating different aspects of the native tendon microenvironment. One approach used to modulate the in vitro microenvironment and enhance extracellular matrix (ECM) deposition is macromolecular crowding (MMC). MMC is based on the addition of inert macromolecules to the culture media mimicking the dense extracellular matrix. In addition, as tendon has been described to be a relatively avascular and hypoxic tissue and low oxygen tension can stimulate collagen synthesis and cross-linking, we venture to assess the synergistic effect of MMC and low oxygen tension on human tenocyte phenotype maintenance by enhancing synthesis and deposition of tissue-specific ECM. Human tendons were kindly provided from University Hospital Galway, after obtaining appropriate licenses, ethical approvals and patient consent. Afterwards, tenocytes were extracted using the migration method. Experiments were conducted at passage three. Optimization of MMC conditions was assessed using 50 to 500 μg/ml carrageenan (Sigma Aldrich, UK). For variable oxygen tension cultures, tenocytes were incubated in a Coy Lab (USA) hypoxia chamber. ECM synthesis and deposition were assessed using SDS-PAGE (BioRad, UK) and immunocytochemistry (ABCAM, UK) analysis. Protein analysis for Scleraxis (ABCAM, UK) was performed using western blot. Gene analysis was conducted using a gene array (Roche, Ireland). Cell morphology was assessed using bright-field microscopy. All experiments were performed at least in triplicate. MINITAB (version 16, Minitab, Inc.) was used for statistical analysis. Two-sample t-test for pairwise comparisons and ANOVA for multiple comparisons were conducted SDS-PAGE and immunocytochemistry analysis demonstrated that human tenocytes treated with the optimal MMC concentration at 2% oxygen tension showed increased synthesis and deposition of collagen type I, the major component of tendon ECM. Moreover, immunocytochemistry for the tendon-specific ECM proteins collagen type III, V, VI and fibronectin illustrated enhanced deposition when cells were treated with MMC at 2% oxygen tension. In addition, protein analysis revealed elevated dexpression of the tendon-specific protein Sclearaxis, while a detailed gene analysis revealed upregulation of tendon-related genes and downregulation of trans-differentiation markers again when cells cultured with MMC at 2% oxygen tension. Finally, low oxygen tension and MMC did not affect the metabolic activity, proliferation and viability of human tenocytes. Collectively, results suggest that the synergistic effect of MMC and low oxygen tension can accelerate the formation of ECM-rich substitutes, which stimulates tenogenic phenotype maintenance. Currently, the addition of substrate aligned topography together with MMC and hypoxia is being investigated in this multifactorial study for the development of an implantable device for tendon regeneration.
Tissue engineering by self-assembly is a technique that consists of growing cells on surfaces made of thermoresponsive polymers, that allow the production of contiguous cell sheets by simply lowering the temperature below the polymer's low critical solution temperature. In this approach cell-cell junctions and deposited extracellular matrix (ECM) remain intact, which provides a better cell localisation at the site of injury. However, these systems lack the possibility to fabricate multi-layered and three-dimensional cell sheets that would better recapitulate native tissues. Moreover, the fabrication of ECM-rich cell sheets would be highly desirable. This limitation could be overcome by inducing macromolecular crowding (MMC) conditions. Herein we venture to fabricate electrospun thermoresponsive nanofibres to sustain the growth and detachment of ECM-rich tissue substitutes in the presence of a MMC microenvironment. A copolymer of 85% poly-N-isopropylacrylamide and 15% N-tert-butylacrylamide (pNIPAAm/NTBA) were used for all experiments. To create aligned nanofibers, the polymer was electrospun and collected on a mandrel rotating at 2000 rpm. Human adipose derived stem cells (hADSC) were treated with media containing macromolecular crowders to enhance matrix deposition. Cell viability and morphology were assessed, and immunocytochemistry was conducted in order to estimate matrix deposition and composition. Adipogenic, osteogenic and chondrogenic assays were performed both with and without the presence of MMC. Non-invasive cell detachment was enabled by decreasing the temperature of culture to 10 °C for 20 minutes. The electrospinning process resulted in the production of pNIPAm/NTBA fibres in the diameter range from 1 to 2 µm and an overall alignment of 80%. Cell viability, proliferation and metabolic activity revealed that hADSCs were able to grow on the thermoresponsive scaffold. The cells were able to detach as an intact cell sheet in presence of MMC. Moreover, it was demonstated that MMC, by a volume extrusion effect, enhances Collagen type I deposition, which is one of the main components of the ECM. Histological analysis revealed that in the presence of MMC the cells were able to self-assembled into three dimensional multi-layers. The cells were able to differentiate towards the osteogenic and adipogenic lineage in the presence of MMC. Interestingly we were able to fabricate three-dimensional chondrogenic cell sheet both with and without MMC. Collectively the pNIPAm/NTBA thermoresponsive fibres were able to sustain the growth and the detachment of ECM-rich multi-layered cell sheets. The pNIPAm/NTBA fibres were able to successfully sustain growth and detachment of ECM-rich tissue equivalents. We believe that replacement, repair and restoration of tissue function can be accomplished best using cells that create their own tissue-specific extracellular matrix with a precision and stoichiometric efficiency still unmatched by man-made devices.
Deep venous thrombosis (DVT) is a potentially serious complication after total hip (THA) and knee (TKA) arthroplasty, traditionally justifying aggressive prophylaxis with low molecular weight heparin (LMWH) or direct oral anticoagulants (DOA) at the cost of an increased risk of bleeding. However, fast-track procedures might reduce the DVT risk and decrease the cost-benefit ratio of the current recommendations. The objective of this study was to compare thrombotic and bleeding risk in an unselected population of elective THA and TKA with a fast-track procedure. A series of 1,949 patients were analyzed prospectively. There were 1,136 women and 813 men, with a mean age of 70 years. In particular, 16% were previously treated by antiplatelet agents and 8% by anticoagulants. All patients followed a fast-track procedure including early walking within 24 hours of surgery, and 80% of patients returned home after surgery, with a mean length of stay of 3 days (THA) or 4 days (TKA). The occurrence of a thromboembolic event or hemorrhagic complication has been identified.Introduction
MATERIAL - METHODS
Prosthetic joint infection (PJI) remains the most severe complication of arthroplasty. Failure of intensive, long-term antibiotic treatment for PJI often requires removal of the implant. Antibiotic failure is thought to be caused by biofilm and persister formation. Novel anti-biofilm and anti-persister strategies are urgently needed. Here, we investigated the effects of several antimicrobial peptides on the bacteria within antibiotic-treated biofilms in an On polystyrene, a mature (7 day-old) methicillin-resistant Aims
Methods
Rotational or axial alignment is an important concept in total knee surgery. Malrotation of the femoral component can lead to patellofemoral maltracking, pain and stiffness. In reconstruction surgery of the knee, achievement of correct rotation is even more difficult because of the lack of anatomical landmarks. The linea aspera is often the only remaining landmark, but its reliability is questionable. Can custom-made 3D-guides help with rotational alignment of the knee after a wide resection of the distal femur?Introduction
Goal of research
Orthopedic implant related surgical site infection (SSI) is a severe complication which represents an important challenge concerning to its treatment. Therefore, gram-negative orthopedic infections have recently become a global concern. Retrospective study through searching of the SCIH (infection control service) database, concerning to the year 2016 and 2017. Cases selected were those of implant placement clean surgeries (osteosynthesis or prosthetic placement) which evolved with SSI and Gram-negative bacterial growth in bone tissue or periprosthetic cultures.Aim
Method
Obese patients are not only more likely to receive total joint arthroplasty, but are also more prone to postoperative complications. The most severe complication is a prosthetic joint infection (PJI), occurring two to four times more often in severely obese patients (BMI ≥ 35kg/m2) compared to non-obese patients. This higher risk for PJI may be attributed to higher glucose levels in case of diabetes mellitus, diminished wound healing or inadequate antibiotic prophylaxis. To ultimately improve the prevention measures for this specific patient category, we aimed to describe the clinical and microbiological characteristics of early acute PJI in severely obese patients. We retrospectively evaluated patients with early acute PJI of the hip and knee treated with DAIR between 2006 and 2016 in three Dutch hospitals. According to protocol, cefazolin was administered as antibiotic prophylaxis during arthroplasty and adjusted to bodyweight. PJI was diagnosed using the criteria described by the Musculoskeletal Infection Society. Early acute PJI was defined as less than 21 days of symptoms and a DAIR performed within 90 days after index surgery. Several clinical and microbiological variables were collected and analyzed. Severe obesity was defined as a BMI ≥ 35kg/m2.Aim
Method
Debridement, antibiotics and implant retention (DAIR) is a widely used treatment modality for early acute prosthetic joint infection (PJI). A preoperative risk score was previously designed for predicting DAIR failure, consisting of chronic renal failure (K), liver cirrhosis (L), index surgery (I), cemented prosthesis (C) and C-reactive protein >115mg/L (KLIC). The aim of this study was to validate the KLIC score in an external cohort. We retrospectively evaluated patients with early acute PJI treated with DAIR between 2006 and 2016 in three Dutch hospitals. Early acute PJI was defined as less than 21 days of symptoms and DAIR performed within 90 days after index surgery. Failure was defined as the need for 1) second DAIR, 2) implant removal, 3) suppressive antimicrobial treatment or 4) infection-related death within 60 days after debridement.Aim
Method
Systemic metal ion monitoring (Co;Cr) has proven to be a useful screening tool for implant performance to detect failure at an early stage in metal-on-metal hip arthroplasty. Several clinical studies have reported elevated metal ion levels after total knee arthroplasty (TKA), with fairly high levels associated with rotating hinge knees (RHK) and megaprostheses1. In a knee simulator study, Kretzer2, demonstrated volumetric wear and corrosion of metallic surfaces. However, prospective in vivo data are scarce, resulting in a lack of knowledge of how levels evolve over time. The goal of this study was to measure serum Co and Cr levels in several types TKA patients prospectively, evaluate the evolution in time and investigate whether elevated levels could be used as an indicator for implant failure. The study was conducted at Ghent University hospital. 130 patients undergoing knee arthroplasty were included in the study, 35 patients were lost due to logistic problems. 95 patients with 124 knee prostheses had received either a TKA (primary or revision) (69 in 55 patients), a unicompartimental knee arthroplasty (7 UKA), a RHK (revision −7 in 6 patients) or a megaprosthesis (malignant bone tumours − 28 in 27 patients) (Fig 1). The TKA, UKA and RHK groups were followed prospectively, with serum Co and Cr ions measured preoperatively, at 3,6 and 12 months postoperatively. In patients with a megaprosthesis, metal ions were measured at follow-up (cross-sectional study design).Introduction
Patients and Methods
Failure of the polyethylene glenoid component is the most common complication of Total Shoulder Arthroplasty (TSA) and accounts for a majority of the unsatisfactory results after this procedure. Nowadays, most of the shoulder prostheses consist of metal on polyethylene bearing components. Repetitive contact between the metal ball and the polyethylene socket produces progressive abrasion of the implant if the moving part is made of polyethylene. Its debris may then lead to an active osteolysis and implant loosening. Failure of the glenoid component is often manifested clinically by pain, loss of function, and the presence of a clunking noise and leads to revision surgery. The use of ceramic balls aims at the reduction of this phenomenon. In many studies regarding knee and hip replacement it has been shown that the use of ceramic on polyethylene (CoP) is more beneficial in terms of polyethylene wear and failure, when compared to metal on polyethylene (MoP). Since a human shoulder is very different from a hip and a knee, it is not a self-centering, neither congruent joint. And its stability is provided by healthy muscles of the rotator cuff. We decided to compare CoP against MoP in semi- force controlled test setup. Where, for a given governing angular motion the translational motion was a function of contact (frictional) forces between the tested couple (humeral head and PE). This is to our knowledge the first study to address in direct comparison wear in TSA in semi force controlled test setup. Up today, there is no test standard for wear testing of TSA. A customised joint simulator was used to create worst-case scenario motion allowing for simulation of the muscles in two perpendicular axes: inferior – superior (I-S) and anterior – posterior (A-P). Were a governing angular motion (GAM) was the abduction – adduction (±30°) in I-S. A system of springs was created so that the I-S translation and the A-P rotation were a result of the GAM. The stiffens of the springs was tuned based on the MoP pair initial kinematic (1000 cycles) to result in: about 2mm I-S translation, and about ±10° A-P rotation. All samples were tested at the same test station in order to obtain maximal repeatability. Axial load was in range of 100N to 750 N. Three articulating couples for each material were tested for total of 2M cycles. Standard midterm gravimetric measurements were conducted at each 0.5 M cycles.Introduction
Materials and methods
External rotation of the femoral component is one factor that favors a satisfactory clinical result. New technologies have been developed to precisely implant the components of a total knee arthroplasty, including computer-assisted surgery (CAS) and patient-specific instruments (PSIs). The aim of this study was to compare the precision of CAS and PSIs when determining the orientation of the femoral component. A total of 65 patients operated on in 2008 with CAS had pre- and post-operative computed tomography (CT) in which the posterior condylar angle (PCA) was measured. The same pre- and post-operative measurements were performed for 27 patients operated on in 2010 with the assistance of PSI. For both populations, the antero-posterior femoral cuts were directed to implant the femoral component 3° of external rotation from the pre-operative posterior condylar line (PCL).Purpose
Methods
(1) to investigate the relationships between the bony contours of the knee and the Popliteus Tendon (PT) in the healthy knee and after implantation of a TKA and (2) to analyze the influence of implant sizing. With an apparently well-sized TKA, the position of the PT during knee flexion is modified compared with the preoperative situation.Goals of the study
Hypothesis
Assessment for and treatment of osteoporosis is recommended following hip fracture. All forms of osteoporosis treatment require an adequate calcium intake and normal vitamin D levels. This study assesses vitamin D levels in patients with hip fractures and describes guidelines on how to manage low vitamin D levels with high dose oral vitamin D3 (cholecalciferol). Circulating 25-hydroxyvitamin D levels were measured in consecutive patients with a hip fracture over an 18 month period. Substitution therapy with high dose oral cholecalciferol was started in 2 selected cohorts; one group received substitution therapy for 3 days, the second group for 7 days.Introduction
Materials and methods
