To investigate the significance of a cluster of cases of glenohumeral chondrolysis occuring following the intra-articular injection of methylene blue to assess rotator cuff integrity during open anterior acromioplasty. All cases of acromioplasty during the period 1999 to 2004 were reviewed to determine the incidence of chondrolysis with and without methylene blue injection. There was a significantly higher incidence of chondrolysis following intra-articular injection of methylene blue. The association of intra-articular methylene blue with chondrolysis has not been previously described in the literature. We conclude that methylene blue should not be used for intra-articular injection

The removal of cement debris at the time of primary and revision joint replacement has been facilitated through the introduction of coloured bone cements. Up to date, few studies have evaluated the effect of methilene blue dye on physical, mechanical and pharmacological properties of cements. In this light, we evaluated the effects of adding methylene blue to bone cement with or without antibiotics (gentamicin, vancomycin or both). The addition of methylene blue to plain cement significantly decreased its mean compression (95.4±3 MPa vs 100.1±6 MPa, p = 0.03) and bending (65.2±5 MPa vs 76.6±4 MPa, p < 0.001) strengths, mean setting time (570±4 seconds vs 775±11 seconds, p = 0.01), as well as its mean elastic modulus (2744±97 MPa vs 3281±110 MPa, p < 0.001). Bending resistance decreases after the supplementation of the coloured cement with vancomycin and gentamicin (55.7±4 MPa vs 65.2±5 MPa, p < 0.001). The release of antibiotics from the bone cement was significantly decreased by the methylene blue. Indeed, the release of gentamicin alone was 385.5±26 μg in comparison to 228.2±24 μg when the methylene blue was added (p < 0.001), while the release of gentamicin in combination with vancomycin was 613±25 μg vs 498.5±70 μg (p = 0.018) when the dye was added to the same formulation. With this study we demonstrated several theoretical disadvantages of the antibiotic-loaded bone cement coloured with methylene blue, although caution should be exercised in transferring our findings to the clinical context. Based on our findings, we do not recommend methylene blue supplementation of PMMA for routine clinical use

Introduction. The aim of this study was to investigate whether methylene blue dye, commonly used in sterile surgical marker pens, would have an effect on human chondrocyte viability, when cultured on a collagen membrane in-vitro. Methods. Bilayered collagen membranes were seeded in duplicate with 12 million human chondrocytes per ml and cultured for 24 hours under standard conditions. Group A consisted of a membrane marked with methylene blue ink on its smooth side, group B marked on its porous side, and group C acting as an unmarked control. At the end of the culture period the membranes were qualitatively analysed for cell survival by live/dead fluorescent staining under confocal microscopy. Results. The control membranes of group C showed normal ‘live’ staining of cells. A marked reduction in chondrocyte viability was seen in groups A and B, with a significant reduction in viability seen opposite to the ink mark on the smooth side in A, and practically no cell viability in the same position on the porous side in group B. Conclusions. Methylene blue dye in surgical marker pen ink has been shown to be cytotoxic to chondrocytes cultured on bilayered collagen membranes. Sterile surgical marker pens are commonly used in cartilage repair surgery to aid in the placement of periosteal patches or collagen membranes in autologous chondrocyte transplantation. Surgeons need to be aware of the potential harmful effects of methylene blue dye when using surgical marker pens in cell based regenerative therapies

Vertebral metastases are the most common type of malignant lesions of the spine. Although this tumour is still considered incurable and standard treatments are mainly palliative, the standard approach consists in surgical resection, which results in the formation of bone gaps. Hence, scaffolds, cements and/or implants are needed to fill the bone lacunae. Here, we propose a novel approach to address spinal metastases recurrence, based on the use of anti-tumour metallic-based nanostructured coatings. Moreover, for the first time, a gradient microfluidic approach is proposed for the screening of nanostructured coatings having anti-tumoral effect, to determine the optimal concentration of the metallic compound that permits selective toxicity towards tumoral cells. Coatings are based on Zinc as anti-tumour agent, which had been never explored before for treatment of bone metastases. The customized gradient generating microfluidic chip was designed by Autodesk Inventor and fabricated from a microstructured mould by using replica moulding technique. Microstructured mould were obtained by micro-milling technique. The chip is composed of a system of microfluidic channels generating a gradient of 6 concentrations of drug and a compartment with multiple arrays of cell culture chambers, one for each drug concentration. The device is suitable for dynamic cultures and in-chip biological assays. The formation of a gradient was validated using a methylene blue solution and the cell loading was successful. Preliminary biological data on 3D dynamic cultures of stromal cells (bone-marrow mesenchymal stem cells) and breast carcinoma cells (MDA-MB-231) were performed in a commercial microfluidic device. Results showed that Zn eluates had a selective cytotoxic effect for tumoral cells. Indeed, cell migration and cell replication of treated tumoral cells was inhibited. Moreover, the three-dimensionality of the model strongly affected the efficacy of Zn eluates, as 2D preliminary experiments showed a high cytotoxic effect of Zn also for stromal cells, thus confirming that traditional screening tests on 2D cultured cells usually lead to an overestimation of drug efficacy and toxicity. Based on preliminary data, the customized platform could be considered a major advancement in cancer drug screenings as it also allows the rapid and efficient screening of biomaterials having antitumor effect

Abstract. The lateral ligaments of the ankle composed of the anterior talofibular (ATFL), calcaneofibular (CFL) and posterior talofibular ligaments (PTFL), are amongst the most commonly injured ligaments of the human body. Although treatment methods have been explored exhaustively, healing outcomes remain poor with high rates of re-injury, chronic ankle instability and pain persisting. The introduction and application of tissue engineering methods may target poor healing outcomes and eliminate long-term complications, improving the overall quality of life of affected individuals. For any surgical procedure or tissue-engineered replacement to be successful, a comprehensive understanding of the complete anatomy of the native structure is essential. Knowledge of the dimensions of ligament footprints is vitally important for surgeons as it guides the placement of bone tunnels during repair. It is also imperative in tissue-engineered design as the creation of a successful replacement relies on a thorough understanding of the native anatomy and microanatomical structure. Several studies explore techniques to describe ligament footprints around the body, with limited studies describing in-depth footprint dimensions of the ATFL, CFL and PTFL. Techniques currently used to measure ligament footprints are complex and require resources which may not be readily available, therefore a new methodology may prove beneficial. Objectives. This study explores the application of a novel technique to assess the footprint of ankle ligaments through a straightforward inking method. This method aims to enhance surgical technique and contribute to the development of a tissue-engineered analogue based on real anatomical morphometric data. Methods. Cadaveric dissection of the ATFL, CFL and PTFL was performed on 12 unpaired fresh frozen ankles adhering to regulations of the Human Tissue (Scotland) Act. The ankle complex with attaching ligaments was immersed in methylene blue. Dissection of the proximal and distal entheses of each ligament was carried out to reveal the unstained ligament footprint. Images of each ligament footprint were taken, and the area, length and width of each footprint were assessed digitally. Results. The collective area of the proximal entheses of the ATFL, CFL and PTFL measures 142.11 ± 12.41mm2. The mean areas of the superior (SB) and inferior band (IB) of the distal enthesis of the ATFL measured 41.72 ± 5.01mm2 and 26.66 ± 3.12mm2 respectively. The footprint of the distal enthesis of the CFL measured 146.07 ± 14.05mm2, while the footprint of the distal PTFL measured 126.26 ± 8.88mm2. The proximal footprint of the ATFL, CFL and PTFL measured 11.06 ± 0.69mm, 7.87 ± 0.43mm and 10.52 ± 0.63mm in length and 8.66 ± 0.50mm, 9.10 ± 0.92mm and 14.41 ± 1.30mm in width on average. The distal footprint of the ATFL (SB), ATFL (IB), CFL and PTFL measured 10.92 ± 0.81 mm, 8.46 ± 0.46mm, 13.98 ± 0.93mm and 11.25 ± 0.95mm in length and 7.76 ± 0.59mm, 7.51 ± 0.64mm, 18.98 ± 1.15mm and 24.80 ± 1.25mm in width on average. Conclusions. This methodology provides an effective approach in the identification of the footprint of the lateral ligaments of the ankle to enhance surgical precision and accuracy in tissue-engineered design. Declaration of Interest. (b) declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported:I declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project

Introduction. Human Mesenchymal stem cells (hMSCs) are a promising source for articular cartilage repair. Unfortunately, under in vitro conditions, chondrogenically differentiated hMSCs have the tendency to undergo hypertrophy similar to growth plate chondrocytes. Retinoic acid (RA) signalling plays a key role in growth plate hypertrophy. Whilst RA agonists block chondrogenesis and foster hypertrophy during later stages, RAR inverse agonists (IA) enhance chondrogenesis when applied early in culture. Therefore, we hypothesized that treatment with RAR IA will attenuate hypertrophy in chondrogenically differentiated hMSCs. To test this hypothesis, we analysed early (initial chondrogenic differentiation) and late treatment (hypertrophy stage) of hMSCs with an RAR IA. Methods. Pellets of passage 2 hMSCs were formed in V-bottom well plates by centrifugation and pre-differentiated in a chemically defined medium containing 10ng/mL TGFß (CM+) for 14 days. Thereafter, pellets were cultured for an additional 14 days under 6 conditions: CM+, CM- (w/out TGFß), and hypertrophic medium (CM- with 25 ng/ml BMP 4, w/out dexamethasone). Each of these first three conditions was additionally supplemented with the RA receptor (RAR) inverse agonist BMS493 (BMS) at 2μM after 14 days of chondrogenic pre-differentiation. One additional BMP4 group was supplemented with BMS from the beginning of chondrogenic differentiation until day 14. The pellets were assessed for gene expression (Col 2, Col 10, Col 1 and MMP13) and histologically using dimethyl methylene blue (DMMB), alkaline phosphatase staining (ALP) and collagen II and X immunohistochemistry. Results. Hypertrophy was reduced by addition of BMS at day 14 and further reduced by addition from the beginning. BMS treatment resulted in smaller cells under hypertrophic conditions, higher collagen II content in chondrogenic groups and reduction in collagen X production and ALP activity in every condition. Gene expression data for hypertrophic markers, collagen X and MMP13, were upregulated under the influence of BMP4 but a distinct downregulation in MMP13 expression was shown upon addition of BMS during the late stage differentiation and further reduced upon addition during early stage chondrogenesis. Furthermore, Collagen X expression was reduced by early BMS treatment. Discussion. The treatment with the RAR IA, BMS, attenuated hypertrophic changes in chondrogenically differentiated hMSCs as demonstrated by histology, immunohistochemistry and PCR. These findings suggest an additional approach to attenuate hypertrophy in chondrogenically differentiated hMSCs. Current studies are exploring the timing and dose of BMS to most efficaciously prevent hypertrophy

The trapeziometacarpal joint (TMJ) is the most commonly involved arthritic joint in the hand and is often injected in the outpatient setting. This study assesses the accuracy of TMJ injections. Six pairs of thawed, fresh-frozen cadaveric upper limbs were placed in the anatomic position. The limbs were randomized to be injected by one of two clinicians (a senior and a junior orthopaedic trainee). The TMJ of these specimens was palpated and injected with 0.5mls aqueous jelly dyed with methylene blue. An independent investigator dissected the specimens and the location of the dye was recorded. A Posterior-Anterior radiograph was then taken to assess the bony anatomy of the joint and graded according to Eaton's classification. Dye was found inside the joint capsule in 10 (83%) of the 12 specimens. Using Fishers Exact test no significant difference was found between the 2 injectors (p=0.46). The 2 joints where the dye was extra-articular had grade III and IV arthritis, whereas all other joints were graded I. This study shows that good accuracy of TMJ injection can be achieved using palpation in the earlier stages of TMJ arthritis, when surface anatomy is accurate enough for an intra-articular injection. This is also when synovitis is more prevalent and injections are more relevant. However the failure rate of injections increases as the disease advances

It is nowadays widely recognized that patient satisfaction following knee arthroplasty strongly depends on ligament balancing. To obtain this balancing, the occurring ligament strain is assumed to play a crucial role. To measure this strain, a method is described in this paper that allows full field 3D evaluation of the strains. The latter is preferred over traditional measurement techniques, e.g. displacement transducers or strain gauges, as human soft tissue is not expected to deform uniformly due to its highly inhomogeneous and anisotropic properties. To facilitate full field strain measurements, the 3D digital image correlation (DIC) technique was adopted. This technique was previously validated by our research group on human tissue. First, a high contrast speckle pattern was applied on the sMCL. Therefore, the specimens are first coated with a small layer of methylene blue. Following, a random white speckle pattern is applied. During knee flexion, two cameras simultaneously take pictures of the deforming region at predefined flexion angles. Using dedicated software, the captured images are eventually combined and result in 3D full field strains and displacements. Using this method, the strain distribution was studied in six cadaveric knees during flexion extension movement. Therefore, the femur was rigidly fixed in a custom test rig. The tibia was left unconstrained, allowing the six degrees of freedom in the knee. A load was applied to all major muscles in physiological directions of each muscle by attaching a series of calibrated weights (Farahmand et al., J Orthop Res., 1998;16(1)). The direction of the pulling cables was controlled using a digital inclinometer for each specimen. As a result, a statically balanced muscle loading of the knee was obtained. From these cadaveric experiments, it is observed that on average the sMCL behaves isometrically between 0° and 90° of flexion. However, high regional differences in strain distribution are observed from the full field measurements. The proximal region of the sMCL experiences relatively high strains upon flexion. These strains are positive (tension) in the anterior part and negative (compression) in the posterior region. In contrast, the distal region remains approximately isometric upon knee flexion (see Figure 1). It is accordingly concluded that the sMCL behaves isometric, though large regional differences are observed. The proximal region experiences higher strains. Furthermore, the DIC technique provided valuable insights in the deformation of the sMCL. This technique will therefore be applied to study the impact of knee arthroplasty in the near future. Caption with figure 1: Full field strain distribution in the sMCL's longitudinal direction for specimen in 45° (a) and 90° (b) of knee flexion

Introduction: Acetabular osteolysis is common behind cups with holes (the reported incidence is 9% to 36%). Fluid pressure has been implicated in the pathogenesis of osteolysis. Aim: To test the hypothesis that a polyethylene liner in a metal cup can act as a pump in vivo. Methods: This study was performed during revision surgery in six cases. The components were from several manufacturers. All were ingrown uncemented cups that had osteolytic lesions associated with holes in the cup. A cannula was placed through the capsule into the hip joint and another was placed through the periosteum and bone of the ilium into the osteolytic lesion above the ingrown cup. The continuity of these two spaces through the holes in the cup was confirmed by the injection of methylene blue. Pressure transducers were then connected to both cannulae. Measurements were taken while applying compression and distraction forces across the artificial hip joint. Results: Compression and distraction loads produced a rise (48mmHg) and a fall (35mmHg) respectively in the pressure in the osteolytic lesion but no change in the hip joint pressure, thereby, demonstrating a pumping action. After exposing the prosthesis we were able to demonstrate 1mm to 2mm of in-out excursion of the polyethylene liner in the metal cup, which may explain the mechanism of the pumping effect. Discussion: The polyethylene liner in the metal shell can act as a pump. Compression and distraction forces, such as occur in normal gait, produce changes in fluid pressure, which are transmitted through the holes and may cause osteolysis behind the cup

Purpose of the study: Surgical reconstruction procedures using the gracilus myocutaneous flap may be compromised if partial or total necrosis of the skin cover develops. The purpose of this study was to describe the anatomic blood supply to the gracilus muscle and the corresponding skin cover in an attempt to better understand the arterial afferences to the skin and define the safest topography for a transferable zone of skin. Material and methods: We dissected the thighs of human adult cadavers to detail the vascular bundles feeding the gracilus muscle. Selective injections of methylene blue were used in the main gracilus pedicle; the area of skin colored was then measured (greatest and smallest diameter) for computation of the surface area using an imaging software. Selective injection of a colored latex fluid enabled description of the perforating vessels between the muscle body and the skin cover. Results: Preliminary results of nine dissections showed that the main blood supply of the gracilus muscle arose from the deep femoral artery (n=8 dissections) or the common femoral artery (n=1 dissection) then penetrated the muscle 90.55 mm below the pubis with a mean diameter of 1.32 mm. The muscle was fed by one to four accessory arteries. The skin cover was stained in all cases, the area involved lying over the proximal and mid thirds of the muscle. The surface area was irregular, the mean length being 127.5 mm and the mean width 91.66 mm. The computed surface area was 88.08 cm. 2. on average (range 58–120.95 cm. 2. ). Each muscle had two to six perforating vessels issuing from the opposite side of the main pedicle and comprised within a 48 mm long segment before dividing at the subcutaneous level. A mathematical model correlated the skin surface area to the number of perforating arteries. Discussion and conclusion: Our findings suggest it would be possible to determine the surface area of skin transferable with a gracilus muscle flap based on high-frequency duplex-Doppler assessment of the number and position of the perforating arising from the muscle and feeding the skin surface. The linear distribution of the surfaces measured as a function of the number of perforating arteries suggests that more reliable conditions for gracilus myocutaneous flap harvesting could be proposed to minimize the risk of cutaneous necrosis

INTRODUCTION. Appropriate, well characterized animal models remain essential for preclinical research. This study investigated a relevant animal model for cancellous bone defect healing. Three different defect diameters of fixed depth were compared in both skeletally immature and mature sheep. This ovine model allows for the placement of four confined cancellous defects per animal. METHODS. Defects were surgically created and placed in the cancellous bone of the medial distal femoral and proximal tibial epiphyses (See Figure 1). All defects were 25 mm deep, with defect diameters of 8, 11, and 14 mm selected for comparison. Defects sites were flushed with saline to remove any residual bone particulate. The skeletally immature and mature animals corresponded to 18 month old and 5 year old sheep respectively. Animals were euthanized at 4 weeks post-operatively to assess early healing. Harvested sites were graded radiographically. The percentage of new bone volume within the total defect volume (BV/TV) was quantified through histomorphometry and μ-CT bone morphometry. Separate regions of interest were constructed within the defect to assess differences in BV/TV between periosteal and deep bone healing. Defect sites were PMMA embedded, sectioned, and stained with basic fuschin and methylene blue for histological evaluation. RESULTS. The animals tolerated the surgery well, with no incidence of fractures within the four weeks. Healing of the defects progressed via endochondral ossification, with none of the defects being completely healed within the 4 week time point. Bone volume fraction (BV/TV) significantly decreased with an increasing defect diameter. Actual bone volume (BV), however, increased with defect diameter, suggesting a correlation between biological response and severity of injury. Three distinct healing regions were found to exist within the defect and along the axis of the defect, with significant differences detected in the BV/TV between adjacent regions. Histologically, the 5 year old animals appeared to have decreased osteoblast activity, and lower osteocyte density within the newly formed woven bone. On occasion, the defects were found to intersect the tibial growth plate in the 18 month old animals, with bone replacing the proliferating chondrocyte zone (See Figure 2). Additionally, the 14 mm defect was not able to be placed in the tibia of sheep due to the possibility of the defect entering the tibial intramedullary (IM) canal, and the lack of cancellous bone between the tibial plateau and IM canal. Both these issues considerably affect this model and should be avoided. CONCLUSION. The surgical placement of 11 mm diameter defects in the proximal tibial and distal femoral epiphyses of skeletally mature sheep presents a suitable large animal model to study early healing of cancellous bone defects. This refined model allows for the placement of four separate non-healing defects within a single sheep, and allows for the possibility to reduce animal numbers required to obtain information

Aims

Steroid injections are used for subacromial pain syndrome and can be administered via the anterolateral or posterior approach to the subacromial space. It is not currently known which approach is superior in terms of improving clinical symptoms and function. This is the protocol for a randomized controlled trial (RCT) to compare the clinical effectiveness of a steroid injection given via the anterolateral or the posterior approach to the subacromial space.

Aims

Staphylococcus aureus is a major cause of septic arthritis, and in vitro studies suggest α haemolysin (Hla) is responsible for chondrocyte death. We used an in vivo murine joint model to compare inoculation with wild type S. aureus 8325-4 with a Hla-deficient strain DU1090 on chondrocyte viability, tissue histology, and joint biomechanics. The aim was to compare the actions of S. aureus Hla alone with those of the animal’s immune response to infection.

Aims

Minimally manipulated cells, such as autologous bone marrow concentrates (BMC), have been investigated in orthopaedics as both a primary therapeutic and augmentation to existing restoration procedures. However, the efficacy of BMC in combination with tissue engineering is still unclear. In this study, we aimed to determine whether the addition of BMC to an osteochondral scaffold is safe and can improve the repair of large osteochondral defects when compared to the scaffold alone.

Aims

Extracellular matrix (ECM) and its architecture have a vital role in articular cartilage (AC) structure and function. We hypothesized that a multi-layered chitosan-gelatin (CG) scaffold that resembles ECM, as well as native collagen architecture of AC, will achieve superior chondrogenesis and AC regeneration. We also compared its in vitro and in vivo outcomes with randomly aligned CG scaffold.

Objectives

An experimental rabbit model was used to test the null hypothesis, that there is no difference in new bone formation around uncoated titanium discs compared with coated titanium discs when implanted into the muscles of rabbits.

The April 2012 Wrist & Hand Roundup³⁶⁰ looks at releasing the trigger finger, function in the osteoarthritic hand, complex regional pain syndrome, arthroscopic ligamentoplasty for the injured scapholunate ligament, self-concept and upper limb deformities in children, wrist arthroscopy in children, internal or external fixation for the fractured distal radius, nerve grafting, splinting the PIPJ contracture, and finding the stalk of a dorsal wrist ganglion