Post-natal vasculogenesis, the process by which vascular committed bone marrow stem cells or endothelial precursor cells migrate, differentiate and incorporate into the nacent endothelium and thereby contribute to physiological and pathological neurovascularisation, has stimulated much interest. Its contribution to neovascularisation of tumours, wound healing and revascularisation associated with ischaemia of skeletal and cardiac muscles is well established. We evaluated the responses of endothelial precursor cells in bone marrow to musculoskeletal trauma in mice. Bone marrow from six C57 Black 6 mice subjected to a standardised, closed fracture of the femur, was analysed for the combined expression of cell-surface markers stem cell antigen 1 (sca-1. +. ) and stem cell factor receptor, CD117 (c-kit. +. ) in order to identify the endothelial precursor cell population. Immunomagnetically-enriched sca-1. +. mononuclear cell (MNC. sca-1+. ) populations were then cultured and examined for functional vascular endothelial differentiation. Bone marrow MNC. sca-1+,c-kit+. counts increased almost twofold within 48 hours of the event, compared with baseline levels, before decreasing by 72 hours. Sca-1. +. mononuclear cell populations in culture from samples of bone marrow at 48 hours bound together Ulex Europus-1, and incorporated fluorescent 1,1′-dioctadecyl- 3,3,3,’3′-tetramethylindocarbocyanine perchlorate-labelled acetylated low-density lipoprotein intracellularily, both characteristics of mature endothelium. Our findings suggest that a systemic provascular response of bone marrow is initiated by musculoskeletal trauma. Its therapeutic manipulation may have implications for the potential enhancement of neovascularisation and the healing of fractures

Myostatin (GDF-8) is known to play an important role in muscle regeneration, and myostatin is also expressed during the early phases of fracture healing. In this study we used fluorescent immunohistochemistry to define the temporal and spatial localization of myostatin during muscle and bone repair following deep penetrant injury in a mouse model. We then used hydrogel delivery of exogenous myostatin in the same injury model to determine the effects of myostatin exposure on muscle and bone healing. Results show that while myostatin was constitutively expressed in the cytoplasm of intact skeletal muscle fibers, a pool of intense myostatin staining was observed amongst injured skeletal muscle fibers 12-24 hours post-surgery. Myostatin was also expressed in the soft callus chondrocytes 4 days following osteotomy. Hydrogel delivery of 10 or 100 ug/ml recombinant myostatin decreased fracture callus cartilage area relative to total callus area in a dose-dependent manner by 41% and 80% (p<0.05), respectively, compared to vehicle treatment. Myostatin treatment also dose-dependently decreased fracture callus total bone volume by 23% and 47% (p<0.05), with the higher dose of recombinant myostatin yielding the greatest decrease in callus bone volume. Finally, exogenous myostatin treatment caused a significant, dose-dependent increase in fibrous tissue formation in skeletal muscle. Together, these findings suggest that myostatin may inhibit bone repair after traumatic musculoskeletal injury through both autocrine (soft-callus chondrocytes) and paracrine (surrounding injured muscle fibers) mechanisms. Thus, early pharmacological inhibition of myostatin is likely to improve the regenerative potential of both muscle and bone following deep penetrant musculoskeletal injury

Fracture related infection remains a major challenge in musculoskeletal trauma surgery. Despite best practice, treatment strategies suffer from high failure rates due to antibiotic resistance and tolerance. Bacteriophages represent a promising alternative as they retain activity against such bacteria. However, optimal phage administration protocols remain unknown, although injectable hydrogels, loaded with phage and conventional antibiotics, may support conventional therapy. In this study we tested the activity of meropenem, and two newly isolated bacteriophages (ϕ9 and ϕ3) embedded within alginate-chitosan microbeads and a hydrogel. Antibiotic and phage stability and activity were monitored in vitro, over a period of 10 days. In vivo, the same material was tested in treatment of a 5-day old Pseudomonas aeruginosa infection of a tibial plate osteotomy in mice. Treatment involved debridement and 5 days of systemic antibiotic therapy plus: i- saline, ii-phages in saline, iii-phages and antibiotics loaded into a hydrogel (n=7 mice/group). To assess the efficacy of the treatments, the infection load was monitored during revision surgery with debridement of the infected tissue after 5,10 and 13 days (euthanasia) by CFU and PFU quantification. In vitro testing confirmed that the stability of meropenem and activity of ϕ9 and ϕ3, was not affected within the alginate beads or hydrogel over 10 days. The in vivo study showed that all mice receiving phages and antibiotics loaded into a hydrogel survived the infection with a reduction of the bacterial load in the soft tissue. Active phages could be recovered from the infected site at euthanasia (10. 4. PFU/g). The hydrogel loaded with bacteriophages and meropenem showed a positive result in locally reducing the infection load indicating a synergistic effect of the selected antimicrobials. Overall, our new strategy shows encouraging results for improving the treatment of antibiotic-resistant biofilm infections that are related to medical implants

Fracture related infections (FRI) are debilitating complications of musculoskeletal trauma surgery that can result in permanent functional loss or amputation. This study aims to determine risk factors associated with FRI treatment failure, allowing clinicians to optimise them prior to treatment and identify patients at higher risk. A major trauma centre database was retrospectively reviewed over a six-year period. Of the 102 patients identified with a FRI (66 male, 36 female), 29.4% (n=30) had acute infections (onset <6 weeks post-injury), 34.3% (n=35) had an open fracture. Open fractures were classified using Gustilo-Anderson (GA) classification (type 2:n=6, type 3A:n=16, type 3B:n=10, type 3C:n=3). Patients with periprosthetic infections of the hip and knee joint, those without prior fracture fixation, soft tissue infections, diabetic foot ulcers, pressure sore infections, patients who died within one month of injury, <12 months follow-up were excluded. FRI treatment failure was defined as either infection recurrence, non-union, or amputation. Lifestyle, clinical, and intra-operative data were documented via retrospective review of medical records. Factors with a P-value of p<0.05 in univariate analysis were included in a stepwise multivariate logistic regression model. FRI treatment failure was encountered in 35.3% (n=36). The most common FRI site was the femoral shaft (16.7%; n=17), and 15.7% (n=16) presented with signs of systemic sepsis. 20.6% (n=21) had recurrent infection, 9.8% (n=10) had non-union, and 4.9% (n=5) required an amputation. The mean age at injury was 49.71 years old. Regarding cardiovascular risk factors, 37 patients were current smokers (36.3%), 31 patients were diabetics (30.4%), and 32 patients (31.4%) were obese (BMI≥30.0). Average follow-up time was 2.37 (range: 1.04-5.14) years. Risk factors for FRI treatment failure were BMI>30, GA type 3c, and implant retention. Given that FRI treatment in 35.3% (36/102) ended up in failure, clinicians need to take into account the predictive variables analysed in this study, and implement a multidisciplinary team approach to optimise these factors. This study could aid clinicians to redirect efforts to improve high risk patient management, and prompt future studies to trial adjuvant technologies for patients at higher risk of failure

Abstract. Objectives. Musculoskeletal injuries are the leading contributor to disability globally, yet current treatments do not offer complete restoration of the tissue. This has resulted in the exploration of novel interventions based on tissue engineering as a therapeutic solution. This study aimed to explore novel collagen sponges as scaffolds for bone tissue engineering as an initial step in the construction of tendon-bone co-culture constructs in vitro. Methods. Collagen sponges (Jellagen, UK), manufactured from Jellyfish collagen were seeded with 10,000 rat osteoblast cells (dROBs) and maintained in culture for 6 days (37°C, 5% CO. 2. ). Qualitative viability was assessed by a fluorescent Calcein-AM live cell stain and quantitively via the CYQUANT cell viability assay (Invitrogen, UK) on days 0, 1, 4 and 6 in culture (n=3 per time point). Digital imaging was also used to assess size and shape changes to the collagen sponge in culture. Results. The collagen sponge biomaterial supported dROB adhesion, viability and proliferation with an abundance of viable cells detected by fluorescent microscopy on day 6. Indeed, the quantitative assessment confirmed that cellular proliferation was evident with increases in fluorescence detected from 517 (± 88) RFU to 8730 ± (2228) RFU from day 0 to 6. In addition, the size of the collagen sponges appeared to decrease over time, indicating contraction of the collagen sponges in culture. Conclusions. This preliminary study has demonstrated that the novel collagen sponges support cellular attachment and proliferation of osteoblasts, and is an important first step in building a bone-tendon construct in vitro. Our future work is focussed on using the osteoblast-seeded sponges in combination with tendon cells, to build a co-culture to represent the bone-tendon interface in vitro. This work has the potential to advance the clinical translation of tissue-engineered tendons to the clinic. 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

Our musculoskeletal system has a limited capacity for repair. This has led to increased interest in the development of tissue engineering strategies for the regeneration of musculoskeletal tissues such as bone, ligament, tendon, meniscus and articular cartilage. This talk will review our attempts to use biomaterials and mesenchymal stem cells (MSCs) to bioprint functional articular cartilage and bone grafts for use in bone and joint regeneration. It will begin by describing how 3D bioprinting can be used to engineer biological implants mimicking the shape of specific bones, and how these bioprinted tissues mature into functional bone organs upon implantation into the body. Next, it will be demonstrated that different musculoskeletal injuries can be regenerated using 3D bioprinted implants, including large bone defects and osteochondral defects. The talk will conclude by describing how we can integrate biomaterials and MSCs into 3D bioprinting systems to engineer scaled-up tissues that could potentially be used regenerate entire diseased joints

Conventional non-steroidal anti-inflammatory drugs (NSAIDs) and newer specific cyclo-oxygenase-2 (cox-2) inhibitors are commonly used in musculoskeletal trauma and orthopaedic surgery to reduce the inflammatory response and pain. These drugs have been reported to impair bone metabolism. In reconstruction of the anterior cruciate ligament the hamstring tendons are mainly used as the graft of choice, and a prerequisite for good results is healing of the tendons in the bone tunnel. Many of these patients are routinely given NSAIDs or cox-2 inhibitors, although no studies have elucidated the effects of these drugs on tendon healing in the bone tunnel. In our study 60 female Wistar rats were randomly allocated into three groups of 20. One received parecoxib, one indometacin and one acted as a control. In all the rats the tendo-Achillis was released proximally from the calf muscles. It was then pulled through a drill hole in the distal tibia and sutured anteriorly. The rats were given parecoxib, indometacin or saline intraperitoneally twice daily for seven days. After 14 days the tendon/bone-tunnel interface was subjected to mechanical testing. Significantly lower maximum pull-out strength (p < 0.001), energy absorption (p < 0.001) and stiffness (p = 0.035) were found in rats given parecoxib and indometacin compared with the control group, most pronounced with parecoxib

Introduction. Orthopaedic trauma surgery is characterised by repetitive, forceful tasks that are physically demanding, thus theoretically increasing the risk of musculoskeletal injuries in these surgeons. The aim of this study is to assess prevalence, characteristics and impact of musculoskeletal disorders among orthopaedic trauma surgeons. Methods. A modified version of the physical discomfort survey was sent to surgeon members of the Orthopaedics Trauma Association (OTA) via e-mail. For data analysis, one-way ANOVA and Fisher Exact test were performed to compare the variables where appropriate. P values<0.05 were considered statistically significant. Results. A total of 86 surgeons completed the survey during the period of data collection. Of the respondents 84.9% were males and more than half were aged between 30–45 years old. The majority of musculoskeletal complaints and disorders were low back pain (29.3%), wrist or forearm tendinitis (18.0%), elbow lateral epicondylitis (15.4%), plantar fasciitis (14.7%). When data was analysed according to number of years in practice the results yielded a significant difference between the groups in both number of regions involved (p<0.05) and number of musculoskeletal disorders (p<0.05), as a higher proportion of these were documented in surgeons practicing for 16–20 years and more than 30 years. Also surgeons working in a private setting (p<0.005), surgeons working in more than one institute (p<0.005), increased number of regions involved (p<0.001) and increased number of musculoskeletal disorders (p<0.001) were significantly more likely to require time-off work. Conclusion. To our knowledge, our study is the first of its kind that shows a high percentage of orthopaedic trauma surgeons sustain occupational injuries some time in their careers. Cost of management and rehabilitation of these injuries, in addition to the amount of missed workdays due to these injuries indicate that these injuries have a significant economic burden on the health-care system

Introduction. Tendinopathies are among the most common musculoskeletal injuries. Nowadays, part of its diagnosis is established through subjective qualitative evaluation of 2D ultrasound (US). This enables limited diagnostic differentiation or therapeutic optimization and has limited added value to diagnosis in an earlier stage. It is generally accepted that extra diagnostic information can be obtained via strain evaluation. The accurate validation of strain estimation is challenging due to the lack of a ground-truth. Therefore we evaluate the repeatability of displacement and strain estimations in the longitudinal direction, using an easy, fast and interactive application to estimate local strain during dynamic loading of the tendon. Materials and Methods. One healthy volunteer laid in a prone position with the foot fixed to an isokinetic device. Three sets of passive movement between −10° plantarflexion and +10° dorsiflexion were performed and repeated the following day. During this, US images with a spatial resolution of 0.02mm × 0.09mm were acquired at a frame-rate of 100Hz. The US system used was the Vevo2100 with a MS250 linear array transducer with a center frequency of 20MHz. After image collection, consecutive pairs of 2D images were registered in a multi-resolution scheme, using an affine and b-spline transformation optimized by the minimization of the sum-of-squared differences, to obtain deformation vector fields. Lastly the interactive application allows local analysis of tissue displacement and strain within selected regions of interest. Mean and standard deviation of the intra- and inter-day relative differences were calculated. Results. The results show a mean intra-day relative difference of 13.71%±4.76% in displacement and of 16.29%±5.17% in strain. For inter-day comparison, the relative difference was 16.98%±14.62% in displacement and was 16%±13.51% in strain. Results show physiologically meaningful and similar strain tendencies when grouping proximal and distal regions. Discussion. This work shows promising preliminary data that suggest that with our method strain and deformation can be measured in a reproducible way using high-frequency US, with little effect of slight variations in acquisition conditions. This brings the application of US based strain estimation in clinical scenarios closer to reality. However, further tests are needed to confirm these conclusions

Crown copyright 2009. Published with the (permission of the Defence Science and Technology Laboratory on behalf of the Controller of HMSO. Introduction. The optimum strategy for the care of war wounds is yet to be established. A need exists to model complex extremity injury, allowing investigation of wound management options. Aim. To develop a model of militarily relevant extremity wounding. Study Design. Laboratory study with New Zealand White Rabbits. Methods. Phase 1. Development of injury. Following induction of general anaesthesia, a muscle belly on the flexor aspect of the forelimb of the rabbit was exposed. This was achieved by creating a fascial tunnel under the belly of flexor carpi ulnaris (FCU). Utilising a custom built drop test rig a high energy, short duration impact was delivered. To replicate casualty evacuation timelines, the animal was maintained under anaesthesia for three hours and recovered. The wound was dressed with saline soaked gauze and supportive bandaging. 48 hrs later, the animal was culled and the muscle harvested for histological analysis. Analgesia was administered once a day. Animals were checked by experienced staff at least twice a day and body temperature recorded by a subcutaneous transponder. Phase 2. Contamination of muscle injury. Sequential animals had inoculums of 1×102/100μl, 1×106/100μl and 1×108/100μl of Staphylococcus aureus administered to the muscle immediately after injury. Animals were recovered from anaesthetic and monitored as per phase 1. Delivery was evaluated by droplet spread and via injection by fine bore needle into the muscle belly. At the 48 hour point, the animals were culled, dressings removed, the muscle harvested and auxiliary lymph nodes sampled. Quantitative microbiological analysis was performed to determine colony forming unit counts (CFU) at 24 hours post-collection. Results. Phase 1. Six animals were exposed to a loading of 0.5kg. Histological analysis demonstrated a consistent injury pattern with 20% of the muscle belly becoming necrotic. Following discussion with subject matter experts this was found to be representative of the nature of injury from ballistic limb trauma and was adopted as standard. Phase 2. Twenty-two animals were exposed to the standardised injury and then inoculated at the prescribed challenge doses and delivery methods. A challenge dose of 1×106/100μl S. aureus delivered by droplet provided the greatest consistency. A group of six animals with an average challenge dose of 3.3×106/100μl yielded growth at 48hrs on average of 9.2×106 CFU. There were no adverse effects on animal welfare throughout, with body temperatures within normal limits at all times. Discussion. The use of rabbits in the investigation of musculoskeletal injury and infection is well established. No study to date however has addressed high energy complex soft tissue wounding, contamination and its optimum management. Considering the current burden of such wounds the need for this question to be answered in a research setting is transparent. This model enables a significant, reproducible, contaminated soft tissue injury to be delivered in vivo. It will allow the investigation of complex wound management options including wound coverage and fracture fixation

Objectives

Platelet-rich plasma (PRP) is being used increasingly often in the clinical setting to treat tendon-related pathologies. Yet the optimal PRP preparations to promote tendon healing in different patient populations are poorly defined. Here, we sought to determine whether increasing the concentration of platelet-derived proteins within a derivative of PRP, platelet lysate (PL), enhances tenocyte proliferation and migration in vitro, and whether the mitogenic properties of PL change with donor age.