Summary Statement

Bone stress fracture triggers a rapid increase in blood flow in association with mast cell production of inducible nitric oxide synthase (iNOS). NOS inhibition blocks the increase in blood flow and reduces woven bone formation needed for stress fracture healing.

Renal Osteodystrophy is a type of metabolic bone disease characterized by bone mineralization deficiency due to electrolyte and endocrine abnormalities. Patients with chronic kidney disease (CKD) are more likely to experience falls and fractures due to renal osteodystrophy and the high prevalence of risk factors for falls. Treatment involves medical management to resolve the etiology of the underlying renal condition, as well as management (and prevention) of pathological fractures. A 66-year-old female patient, with severe osteoporosis and chronic kidney disease undergoing haemodialysis, has presented with multiple fractures along the years. She was submitted to bilateral proximal femoral nailing as fracture treatment on the left and prophylactically due to pathological bone injury on the right, followed by revision of the left nail with a longer one after varus angulation and fracture distal to the nail extremity. Meanwhile, the patient suffered a pathological fracture of the radial and cubital diaphysis and was submitted to conservative treatment with cast, with consolidation of the fracture. Posteriorly, she re-fractured these bones after a fall and repeated the conservative treatment. Clinical management: There is a multidisciplinary approach to manage the chronic illness of the patient, including medical management to resolve the etiology and consequences of her chronic kidney disease, pain control, conservative or surgical fracture management and prevention of falls. The incidence of chronic renal disease is increasing and the patients with this condition live longer than previously and are more physically active. Thus, patients may experience trauma as a direct result of increased physical activity in a setting of weakened pathologic bone. Their quality of life is primarily limited by musculoskeletal problems, such as bone pain, muscle weakness, growth retardation, and skeletal deformity. A multidisciplinary approach is required to treat these patients, controlling their chronic diseases, managing fractures and preventing falls

An isolated avulsion fracture of the peroneus longus tendon is seldom seen and potentially can go undiagnosed using basic imaging methods during an initial emergency visit. If not managed appropriately it can lead to chronic pain, a reduced range of motions and eventually affect mobility. This article brings to light the effectiveness of managing such injuries conservatively. A 55 year old postman presented to clinic with pain over the instep of his right foot for 2 months with no history of trauma. Clinically the pain was confined to the right first metatarsophalangeal joint with occasional radiation to the calf. X-ray films did not detect any obvious bony injury. MR imaging revealed an ununited avulsion fracture of the base of the 1st metatarsal. The patient was subsequently injected with a mix of steroid and local anesthetic injections at the painful nonunion site under fluoroscopic guidance. Post procedure there was no neurovascular deficit. The patient was reviewed at three months and his pain score and functional outcome improved significantly. Moreover following our intervention, the Manchester Oxford Foot Questionnaire reduced from 33 to 0. At the one year follow up he remained asymptomatic and was discharged. The peroneus longus tendon plays a role in eversion and planter flexion of foot along with providing stabilization to arches of foot. The pattern of injury to this tendon is based on two factors one is the mechanism of insult, if injured, and second is the variation in the insertion pattern of peroneus longus tendon itself. There is no gold standard treatments by which these injuries can be managed. If conservative management fails we must also consider surgery which involves percutaneous fixation, or excision of the non-healed fracture fragment and arthrodesis. To conclude isolated avulsion fractures of peroneus longus tendon are rare injuries and it is important to raise awareness of this injury and the diagnostic and management challenges faced. In this case conservative management was a success in treating this injury however it is important to take factors such as patient selection, patient autonomy and clinical judgement into account before making the final decision

Abstract. Objectives. The aim of this study was to test the hypothesis that there are two different mallet injuries; specifically, tendinous ones are primarily low energy avulsion injuries whilst bony ones are primarily high-energy hyper-extension injuries. Methods. We reviewed in detail the demographics, mechanisms of injury, concomitant injuries and the radiological findings of patients presenting with bony and tendinous mallet deformities. The sizes of the bony fragments and angulation of the mallet deformities were measured on the initial radiographs using an established technique. Results. There were 62 tendinous mallet injuries (62 patients). These were mostly low energy injuries in men with a mean age of 57 years affecting primarily the middle and ring fingers. They were rarely associated with other injuries. There were 85 bony mallet injuries (83 patients). These were all high energy injuries also mostly in men with a mean age of 40 years primarily affecting the ring and little fingers. There were two double injuries and seven other concomitant injuries in the patients with bony mallet injuries. The extensor lags were a mean of 300 (range 3–590) for the tendinous injuries and 130 (range 0–380) for the bony injuries (p<0.00001). The fracture fragments were a mean of 51 (range 24–80) %; there was no correlation between fragment size and extensor lag. Conclusions. This study and review of the literature further confirm the substantial differences between tendinous and bony mallet injuries. In particular tendinous mallet injuries are avulsion injuries whilst bony mallet injuries are dorsal impaction fractures; they should be assessed and treated differently. Bony mallet injuries may be multiple and may be associated with other injuries. Low energy mallet deformities do not need radiographs

Introduction. The stability of the elbow joint following an acute elbow dislocation is dependent on associated injuries. The ability to identify these concomitant injuries correctly directs management and improves the chances of a successful outcome. Interpretation of plain radiographs in the presence of either a dislocation or post-reduction films with plaster in-situ is difficult. This study aimed to assess the ability of orthopaedic registrars to accurately identify associated bony injuries on initial plain radiographs using CT as the gold standard for comparison. Methods. Patients over the age of 16 years undergoing an elbow CT scan within one week of a documented elbow dislocation between 1st June 2010 and 1st June 2014 were included in the study. Three orthopaedic registrars independently reviewed both the initial dislocation and immediate post reduction plain radiographs to identify any associated bony injuries. This radiograph review was repeated by each registrar after two weeks. The incidence of associated injuries as well as the inter- and intra-observer variability was calculated. Results. 28 patients were included in the study. 54% of the patients were female and the mean age was 45 years (range 16 to 90 years). The incidence of a radial head fracture was 54%, coronoid fracture 43% and epicondyle avulsion 18% on CT. The inter-observer reliability was only shown to be fair amongst registrars and the intra-observer variability moderate. Conclusions. Computerised tomography is a useful adjunct in the assessment of associated osseous injuries following an elbow dislocation due to the presence of a high number of injuries. Plain radiographs alone have been shown to have only a fair and moderate inter and intra-observer variability respectively, therefore a low threshold to obtain further 3D imaging should be practised. Level of Evidence. IV

Abstract. 3D printing of synthetic scaffolds mimicking natural bone chemical composition, structure, and mechanical properties is a promising approach for repairing bone injuries. Direct ink writing (DIW), a type of 3D printing, confers compatibility with a wide range of materials without exposing these materials to extreme heat. Optimizing ink properties such as filament formation capabilities, shear-thinning, and high storage modulus recovery would improve DIW fabrication characteristics. In this study, composite inks based on biodegradable polycaprolactone (PCL), reinforced with nano-hydroxyapatite (HAp), and loaded with vancomycin were designed and evaluated for their rheological properties, wettability, mechanical properties, and antimicrobial properties. The formulated composite inks displayed a shear-thinning behaviour exhibited storage modulus recovery percentages above 80% for all formulations, which is essential for extrusion deposition by DIW at room temperature. Ink formulations were able to form fully interconnected lattice scaffolds with porosities ranging from 42% to 65%. Increasing the HAp concentrations from 55% to 85% w/w increased the shear thinning behaviour and reduced the printed filament width to more closely match the nozzle diameter; this indicates higher HAp proportion reduces ink shrinkage. The scaffold had high wettability at HAp proportions above 65% w/w and the compressive elastic modulus of DIW printed scaffolds exhibited within the range of trabecular bone. Antimicrobial activity was apparent from the agar diffusion assay; zones of inhibition ranging from 15.82 ± 0.25 mm and 20.06 ± 0.25 mm were observed after 24 hr for composite scaffolds loaded with 3% and 9% w/w vancomycin respectively. Vancomycin-loaded PCL/HAp composite inks were developed, displaying good printability, wettability, mechanical properties, and antimicrobial properties, making them an attractive choice for bone repair and regeneration. 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

The stem cell fraction of a cell population is finely tuned by stimuli from the external microenvironment. Among these stimuli, a decrease of extracellular pH (pHe) may occur in a variety of physiological and pathological conditions, including hypoxia and inflammation. Also in bone, the maintenance of acid-base balance is fundamental for skeleton homeostasis. Bone cells are extremely sensitive to the effects of interstitial pH. Acidosis inhibits mineral deposition by osteoblasts and activates osteoclast-mediated bone resorption. Moreover, acidosis is associated with inflammation, and in case of bone injury, local short-term acidosis is a crucial regulator of the healing process. Evidence of the role of acidosis as an enhancer of MSC stemness and for their activation as sensors and switcher of inflammation will be discussed

While new biomaterials for regenerative therapies are being reported in the literature, clinical translation is slow. Existing regenerative approaches rely on high doses of growth factors, such as BMP-2 in bone regeneration, which can cause serious side effects. We describe an ultra-low-dose growth factor technology yielding high bioactivity based on a simple polymer, poly (ethyl acrylate) (PEA), and report its translation to a clinical veterinary setting. This polymer-based technology triggers spontaneous fibronectin organization and stimulates growth factor signaling, enabling synergistic integrin and BMP-2 receptor activation in mesenchymal stem cells. To translate this technology, we use plasma-polymerized PEA on 2D and 3D substrates to enhance cell signaling in vitro, showing the complete healing of a critical-size bone injury in mice in vivo. We demonstrate its safety and efficacy in a Münsterländer dog with a non-healing humerus fracture, establishing the clinical translation of advanced ultra-low-dose growth factor treatment

Bone formation proceeds through two distinct processes. One involves the deposition of bone by osteoblasts (intramembranous ossification) and another through the remodeling of an intermediate cartilaginous matrix formed by chondrogenic differentiation of mesenchymal stem/stromal cells (MSCs) aggregates – a process called endochondral ossification (EO). EO is responsible for formation of long bones during development and most prevalent during facture repair upon callus formation. In adult bone injuries MSCs from periosteum form bone via EO whereas MSCs from bone marrow are primarily differentiate to osteoblast in vivo. We hypothesized that the unique biophysical and biochemical properties of bone mineral phase has a role in programming MSCs. Using a biomimetic bone like apatite (BBHAp) as surrogate for bone mineral phase, we studied the chondrogenic differentiation of human marrow derived MSCs and observed that the BBHAp dictates MSCs fate and strictly dictates the pathway of bone formation in vivo. Through exhaustive dissection of the signaling pathways at play, a prominent role of PTH1R in modulating the effects imposed by the BBHAp has been unraveled. These fundamental insights gained in how bone microenvironment might alter fate of MSCs has important implications for bone repair and regeneration therapies

Introduction. Civilian fractures have been extensively studied with in an attempt to develop classification systems, which guide optimal fracture management, predict outcome or facilitate communication. More recently, biomechanical analyses have been applied in order to suggest mechanism of injury after the traumatic insult, and predict injuries as a result of a mechanism of injury, with particular application to the field so forensics. However, little work has been carried out on military fractures, and the application of civilian fracture classification systems are fraught with error. Explosive injuries have been sub-divided into primary, secondary and tertiary effects. The aim of this study was to 1. determine which effects of the explosion are responsible for combat casualty extremity bone injury in 2 distinct environments; a) in the open and b) enclosed space (either in vehicle or in cover) 2. determine whether patterns of combat casualty bone injury differed between environments Invariably, this has implications for injury classification and the development of appropriate mitigation strategies. Method. All ED records, case notes, and radiographs of patients admitted to the British military hospital in Afghanistan were reviewed over a 6 month period Apr 08-Sept 08 to identify any fracture caused by an explosive mechanism. Paediatric cases were excluded from the analysis. All radiographs were independently reviewed by a Radiologist, a team of Military Orthopaedic Surgeons and a team of academic Biomechanists, in order to determine the fracture classification and predict the mechanism of injury. Early in the study it became clear that due to the complexity of some of the injuries it was inappropriate to consider bones separately and the term ‘Zone of Insult’ (ZoI) was developed to identify separate areas of injury. Results. 62 combat casualties with 115 ZoIs (mean 1.82 zones) were identified in this study. 34 casualties in the open sustained 56 ZoIs (mean 1.65); 28 casualties in the enclosed group sustained 59 ZoIs (mean 2.10). There was no statistical difference in the mean ZoIs per casualty in the open vs enclosed group (Student t-test, p=0.24). Open fractures were more prevalent in the open group compared to the enclosed group (48/59 vs 20/49, Chi-squared test p<0.001). Of the casualties in the open, 1 zone of injury was due to the primary effects of blast, 10 a combination of primary and secondary blast zones, 23 due to secondary effects and 24 from the tertiary effects of blast. In contrast, there were no primary or combined primary and secondary blast zones and only 2 secondary blast zones in the enclosed group. Tertiary blast effects predominated in the enclosed group, accounting for 96% of injury zones (57/59). Analysis of the pattern of injury revealed that there were a higher proportion of lower limb injuries in the Enclosed group (54/59) compared to the Open group (40/58, Chi-squared p<0.05). In the Open group the mechanism of lower limb injury was more evenly distributed amongst mixed primary and secondary blast effects (10), secondary (10) and tertiary (20). In the enclosed group, lower limb injuries were almost exclusively caused by tertiary blast effects (47/48). A similar pattern was also seen in the Upper limb with 4/5 in the enclosed group was injured by tertiary effects compared to 4/18 in the Open Group. In the open group fragmentation injury was the predominant cause of injury (13/18). Conclusions. This data clearly demonstrates two distinct injury groups based upon the casualties' environment. The enclosed environment afforded by buildings and vehicles appears to mitigate the primary and secondary effects of the explosion. However, tertiary blast effects were the predominant mechanism of injury, with severe axial loading to the lower extremity being a characteristic of the fractures seen. In contrast, secondary fragments from the explosion were more likely to result in fractures of casualties caught in the open. The development of future mitigation strategies must be focused on reducing all the different mechanisms of injury caused by an explosion. This will require a better understanding on the effects of bone in high strain environments. This method of forensic biomechanics involving clinicians and engineers, combined with accurate physical and numerical simulations can form the basis in reducing the injury burden to the combat soldier

Complement C5a receptor 1 (C5aR1) has crucial functions in host defense against danger molecules, as does toll-like receptor 2 (TLR2). Both innate immunity receptors interact in immune cells in the context of infectious inflammatory diseases often associated with bone loss, such as periodontitis. C5aR1 plays an important role in bone, as it is expressed on bone cells and strongly upregulated due to bone injury. Importantly, C5aR1-ko mice are protected against arthritis and C5aR1 contributes to bone loss in periodontitis. In contrast, less data exist on the role of TLR2 on osteoblasts, however, it is known that TLR2 is expressed on osteoblasts and contributes to bacterial-induced bone resorption. The aim of this study was to evaluate the interaction of C5aR1 and TLR2 in osteoblasts, including intracellular signaling pathways and gene expression patterns. Primary osteoblasts were isolated from 8–12 week-old WT mice and differentiated for 14 days. Osteoblasts were assessed for expression of C5aR1 and TLR2. Phosphorylation of mitogen-activated protein kinases (MAPK) in response to C5a and Pam3CSK4 (TLR2 agonist) was analyzed by immunoblotting. Gene expression profiling after 30 min and 4 h stimulation of C5a was performed by microarray and candidate genes were validated by quantitative Real-Time PCR (qRT-PCR). Immunoprecipitation was performed using a C5aR1-antibody and C5aR1 and TLR2 were subsequently detected by immunoblotting. Statistics: One way ANOVA p<0.05, n=4–6. We showed that C5aR1 and TLR2 are expressed on osteoblasts and strongly upregulated during differentiation. Via immunoprecipitation, we could show that C5aR1 and TLR2 do physically interact in osteoblasts. We then examined if C5aR1 and TLR2, besides their physical interaction, also act via the same intracellular signaling pathways. Gene expression profiling upon C5a stimulation revealed that the top regulated pathways are related to MAPK and transforming growth factor beta (TGF-β). Respective genes, such as TGF-β (Tgfb1) and its receptor (Tgfbr) were found to be upregulated, and negative MAPK regulators were found to be downregulated, both by microarray analysis and qRT-PCR. Accordingly, we saw a C5aR1- and TLR2-dependent phosphorylation of p38 MAPK. Interestingly, this effect was enhanced and prolonged by costimulation of both receptors. An additive effect of C5aR1 and TLR2 was also seen regarding Cxcl10 levels, which were enhanced compared to C5aR1 or TLR2 stimulation alone. This study shows that C5aR1 and TLR2 interact in osteoblasts, not only physically but also functionally, regarding downstream signaling and target genes. Those data strongly imply a synergistic interplay between the receptors, through which osteoblasts possibly contribute to inflammatory reactions affecting bone

Introduction. Open fractures occur with an annual incidence of 11.5 per 100,000 (6900 pa in UK). Infection rates, even with intravenous broad-spectrum antibiotics, remain as high as 22%. For this reason necessary bone grafting is usually delayed until soft-tissue cover of the bone injury is achieved. A biodegradable bone graft that released sustained high concentrations of antibiotics and encouraged osteogenesis, that could be implanted safely on the day of injury would reduce infection rates and avoid reoperation and secondary grafting. The non –union rate (approx 350 pa in UK) should also be reduced. Such a graft, consisting of a PLA/PGA co –polymer and containing antibiotics, is under development and here we report assessment of spectrum and duration of antimicrobial activity and effect of addition of antibiotics on mechanical properties. Methods. Varying concentrations of gentamicin, colistin, clindamycin and trimethoprim, singly and in combination, were added to the copolymer and test pieces were made. These were then tested using an established method (SPTT) which determines degree and duration of antimicrobial activity as well as risk of emerging resistance. Test bacteria were Staphylococcus epidermidis, Staphylococcus aureus, MRSA and Escherichia coli. Mechanical properties (compressive strength and porosity) were determined using established methods. Results. A combination of gentamicin (4%w/w) and clindamycin (2.5% w/w) gave best results, with inhibitory activity persisting for over 21 days (the target duration) without emergence of resistance. No significant effect of this combination/concentration on mechanical properties was found. Conclusions. The experimental PLA/PGA scaffold containing antibiotics showed activity against the common pathogens of open fractures for a period considered long enough to eradicate contamination acquired at or soon after trauma. At the optimum concentration, they had no significant effect on mechanical properties. In vivo performance is currently being investigated in a sheep model

Tissue reconstruction, based on stem cell activity has become an important part of orthopaedic practice. It is now possible to develop cell lines which are able to produce the fundamental cells which can be used in musculoskeletal regeneration, especially in fracture healing, cartilage regeneration, and muscle repair. However, for the newly implanted cells to be effective, it is vital to have an adequate and developing blood supply to that area. Human and animal studies have demonstrated the marked contribution of bone marrow derived precursor cells in the normal bone healing process. Studies of the application of bone marrow graft have shown that there is greater bone growth when more precursor cells are grafted and these cells are thought to be a mixed population of stems cells and their associated progeny. CD34+ cells have shown remarkable ability to differentiate into many cells types which include chondrocytes and osteocytes. They have also been shown to home on to sites of bone injury and mature into bone cells which take part in the repair process. Colleagues in our laboratories have described a plastic adherent sub-population of CD34+ cells which have been able to reconstitute and sustain hematopoeisis over 5 weeks, similar to long-term marrow culture. This sub-population of cells are called omnicytes. Using this sub-population, we have conducted in vitro and animal experiments using a fracture healing model to assess the role of stem cells in accelerating the fracture healing process. However, it is clear that in order for these cells to be effective, the blood supply needs to be viable. In this paper, the importance of the blood supply and the role of blood flow will be discussed particularly in relation to fracture healing and intervertebral disc regeneration. In fracture healing, the increase of blood flow occurs within the first 6 weeks after the fracture has occurred and CD34+ cells applied to the fracture site via the nutrient artery could accelerate the process of fracture union. In the same way, intervertebral disc patients with chronic low back pain for more than 3 months could be treated with enhanced CD34+ cells in order to allow disc cartilaginous type cells to regenerate. This will be a review of the role of the blood supply, the development of CD34+ cells (namely omnicytes), and the clinical application of these cells to patients with long bone fractures and low back pain

The stress response to trauma is the summation of the physiological response to the injury (the ‘first hit’) and by the response to any on-going physiological disturbance or subsequent trauma surgery (the ‘second hit’).

Our animal model was developed in order to allow the study of each of these components of the stress response to major trauma. High-energy, comminuted fracture of the long bones and severe soft-tissue injuries in this model resulted in a significant tropotropic (depressor) cardiovascular response, transcardiac embolism of medullary contents and activation of the coagulation system. Subsequent stabilisation of the fractures using intramedullary nails did not significantly exacerbate any of these responses.

There is no diagnostic, non-invasive method for the early detection of loosening after total hip arthroplasty. In a pilot study, we have analysed two serum markers of bone remodelling, procollagen I C-terminal extension peptide (PICP) and cross-linked N-terminal telopeptide (NTx), as well as the diagnostic performance of NTx for the assessment of osteolysis. We recruited 21 patients with loosening (group I), 18 with a well-fixed prosthesis (group II) and 17 at the time of primary arthroplasty for osteoarthritis (OA) (group III). Internal normal reference ranges were obtained from 30 healthy subjects (group IV).

The serum PICP level was found to be significantly lower in patients with OA and those with loosening, when compared with those with stable implants, while the NTx level was significantly increased only in the group with loosening, suggesting that collagen degradation depended on the altered bone turnover induced by the implant. This hypothesis was reinforced by the finding that the values in the pre-surgery patients and stable subjects were comparable with the reference range of younger healthy subjects.

A high specificity and positive predictive value for NTx provided good diagnostic evidence of agreement between the test and the clinical and radiological evaluations. The NTx level could be used to indicate stability of the implant. However, further prospective, larger studies are necessary.