INTRODUCTION. In order to address high failure rates following rotator cuff repairs, a greater understanding is required of the underlying structural changes so that treatments can be appropriately targeted and biomarkers of failure can be identified. As collagen is the primary constituent of tendon and determines force transmission, collagen structural changes may affect responses to loading. For example changes in collagen 1 and 5 are associated with the hyperelastic Ehlers-Danlos syndrome, which is diagnosed by looking for pathopneumonic altered collagen fibres or ‘collagen flowers’ in skin using transmission electron microscopy (TEM). To date no study has been performed on the microstructure of torn human rotator cuff tendons using TEM. It was hypothesized that normal, small and massive human rotator cuff tendons tears will have altered microscopic structures. The unique study aimed to use TEM to compare the ultrastructure of small and massive rotator cuff tears, to normal rotator cuff tendons. METHODS. Samples from 7 human rotator cuff tendons repairs were obtained, including 4 massive (>5 cm) and 3 small (< 1 cm) tears, and 3 matched normal controls with no history of connective tissue disorders. Specimens were fixed in 4% glutaraldehyde in 0.1M phosphate buffer, processed and examined blind using routine TEM examination. To assess whether changes in the relative expression of collagen 1 and 5 (COL1A1, COL5A1 and COL5A2) occurred in all tears, qPCR was performed on another 6 phenotypically matched patients. RESULTS. The basic structure of the normal tendon consisted of tightly packed clumps of dense packed parallel running collagen fibers with few fibroblasts and small amounts of fine filamentous material between clumps. In contrast, torn samples were more variable with areas of less dense packing of collagen fibers and larger areas of filamentous material plus variable numbers of lipid droplets both within the fibroblast and between the collagen bundles. There was also evidence of twisting and random orientation of individual collagen fibers. All torn tendons showed evidence of a proportion of the fibers within the collagen bundles being enlarged with a serrated outline, similar in appearance to ‘collagen flowers’. Clear differences between the small and massive tears were not identified. qRT-PCR of torn rotator cuff tendon specimens demonstrated no altered collagen expression compared to normal tendons. DISCUSSION. This novel study has identified the previously unreported presence of atypical collagen fibers with focal swelling resulting in the appearance of ‘collagen flowers’ in torn rotator cuff tendons only. This appearance is considered pathognomonic of Ehlers-Danlos syndrome, classical type 1 and 2. Torn tendons also showed an increase in filamentous material, and infiltration with fat droplets. These novel findings may offer insight into the mechanisms of structural damage that contribute to rotator cuff failure. Further examination is required, to evaluate the significance of these observations

Introduction. The purpose of this study was to clarify the incidence of steroid-induced osteonecrosis among different collagen diseases and to evaluate the predictive factors for steroid-induced osteonecrosis in a prospective MRI study. Methods. We prospectively used MRI to study 337 eligible collagen disease patients requiring corticosteroid therapy and succeeded in examining 1199 joints (hips and knees) in 302 patients with MRI for at least one year starting immediately after the onset of corticosteroid therapy, a one-year follow-up rate of approximately 90%. The underlying collagen diseases included systemic lupus erythematosus (SLE) in 687 joints and a variety of other collagen diseases in 512 joints. Results. Incidence of osteonecrosis was significantly higher in SLE patients than in non-SLE patients (37% versus 21%, p=0.001). Logistic regression analysis revealed that adolescent and adult patients had a significantly higher risk of osteonecrosis compared with pediatric patients (odds ratio [OR]=13.2), that high daily corticosteroid dosage (more than 40 mg/day) entailed a significantly higher risk of osteonecrosis compared with the dosage less than 40 mg/day (OR=4.2), that SLE patients had a significantly higher risk of osteonecrosis compared with non-SLE patients (OR=2.6), and that male patients had a significantly higher risk of osteonecrosis compared with female patients (OR=1.6). Conclusion. These findings suggest that the incidence of steroid-induced osteonecrosis is different among underlying collagen diseases

Introduction. The pathophysiology of high failure rates following rotator cuff tendon repairs, particularly massive tears, is not fully understood. Collagen structural changes have been shown to alter tendon thermal and mechanical properties. Thermal changes in small biopsies, detected by differential scanning calorimetry (DSC) can help to quantify collagen structural differences in torn rotator cuff tendons. This study aimed to form a quantitative rather than qualitative assessment, of whether differences in collagen structure and integrity existed between small biopsies of normal, small and massive rotator cuff tears using DSC. Methods. Thermal properties were measured for 27 human biopsies taken intra-operatively from normal, small, and massive rotator cuff tendon tears. 3 samples were taken from each patient and subjected to a modulated temperature ramp between 20–80°C at a rate of 2°C per minute with 0.318°C amplitude. The melting temperature (TM) is proposed to represent amide-amide hydrogen bond breakage and resulting protein backbone mobility. Denaturing temperature (TD) reportedly corresponds to the temperature at which the proteins fall out of solution. Denaturation enthalpy (H) should correlate with the amount of triple helical structure. Based upon a pre-study power calculation, this study had 90% power to detect a 10% difference in melting and denaturation temperature between groups with alpha=0.05. 1 specimen per patients was also frozen and cryosectioned and polarised light microscopy was used for quantitative validation. The effect of tear size on heat related parameters were performed using a one-way ANOVA test. A student's unpaired t-test was used to search for differences between individual groups (small tears, massive tears and normal tendons). Results. Small and massive rotator cuff tears had significantly higher melting temperature (TM), and denaturation enthalpy (H) compared to controls. The denaturing temperature (TD) was higher in the massive tears only compared to normal tears. No difference was detected between small and massive tears. Histology of massive tendon tears confirmed greater collagen structural disruption compared to small tears and controls. Conclusion. These novel findings suggest greater quantifiable collagen structural disruption in rotator cuff tears, compared to controls. A decrease in important thermal properties of torn tendons suggests that the material is intrinsically less stable. It is likely that torn tendons cannot withstand changes in temperature or stress as well as a perfect material could, particularly for massive tears which are more amenable to denaturation. This study offers insight into possible mechanisms for, or adaptation to, failure in tears and reduced strength

Several bisphosphonates are now available for the treatment of osteoporosis. Porous hydroxyapatite/collagen (HA/Col) composite is an osteoconductive bone substitute which is resorbed by osteoclasts. The effects of the bisphosphonate alendronate on the formation of bone in porous HA/Col and its resorption by osteoclasts were evaluated using a rabbit model. Porous HA/Col cylinders measuring 6 mm in diameter and 8 mm in length, with a pore size of 100 μm to 500 μm and 95% porosity, were inserted into a defect produced in the lateral femoral condyles of 72 rabbits. The rabbits were divided into four groups based on the protocol of alendronate administration: the control group did not receive any alendronate, the pre group had alendronate treatment for three weeks prior to the implantation of the HA/Col, the post group had alendronate treatment following implantation until euthanasia, and the pre+post group had continuous alendronate treatment from three weeks prior to surgery until euthanasia. All rabbits were injected intravenously with either saline or alendronate (7.5 μg/kg) once a week. Each group had 18 rabbits, six in each group being killed at three, six and 12 weeks post-operatively. Alendronate administration suppressed the resorption of the implants. Additionally, the mineral densities of newly formed bone in the alendronate-treated groups were lower than those in the control group at 12 weeks post-operatively. Interestingly, the number of osteoclasts attached to the implant correlated with the extent of bone formation at three weeks. In conclusion, the systemic administration of alendronate in our rabbit model at a dose-for-weight equivalent to the clinical dose used in the treatment of osteoporosis in Japan affected the mineral density and remodelling of bone tissue in implanted porous HA/Col composites

INTRODUCTION. The purpose of this study is to report results from a prospective multicenter study of a bioresorbable type I collagen scaffold used to replace tissue loss following irreparable lateral meniscus injuries. METHODS. 49 non-consecutive patients (33M/16F; mean age 30.5 yrs, range 14.7–54.7 yrs) with irreparable lateral meniscus tears or loss requiring surgical treatment were prospectively enrolled at one of 7 EU centers. 11 patients (22%) had acute injuries of the lateral meniscus, while 38 (78%) had prior surgeries to the involved meniscus. Implantation of the LCMI (now Lateral Menaflex) was performed arthroscopically using an all-inside suturing technique (FASTFIX) combined with inside-out sutures in the more anterior meniscus aspect. Forty-three patients were evaluated with a 2 to 4-year follow-up (FU); mean FU duration was 45 months (range, 33–53 m). Patients were evaluated clinically and by self-assessment using Tegner activity and Lysholm function scores, as well as the Visual Analog Scales (VAS) for pain, and a satisfaction questionnaire. Evaluations were performed pre-operatively, 6 months, 1 year, 2 and 4 years after surgery. X-ray and/or MR-images were taken pre-operatively, and at 1 year and 2 years after surgery. RESULTS. At 3 months after surgery, all patients were able to return to activities of daily living without limitation. Post-op. mean values of all evaluated patients showed statistically significant improvement compared to the preoperative scores. Mean Tegner scores increased from 3.0 to 5.2 at 4 years (0.8 points less than the pre-injury “recall” value); mean Lysholm improved from 63 to 91; mean pain (VAS) decreased from 36 to 8. At the 4-year time point, 86% of the patients stated that they were satisfied with their results (compared to 78% at the one year FU time point). Function and pain scores improved continuously with the highest score at the latest FU evaluation. All data were statistically significant (p<0.001, except for Tegner with p=0.03). MRI examination revealed no changes to the articular cartilage and joint space; however, the newly formed tissue did not present a signal consistent with fully mature native meniscus tissue. Reoperations were necessary in 5 patients (10%) during the FU time period: 3 of the reoperations were for persistent swelling and pain, which were classified as related to the device (6%) and were treated by synovectomy and debridement. Patients recovered without sequelae. The re-op. rate in this series is comparable to re-op. rates reported after lateral meniscal repair. DISCUSSION. Based on available results with a minimum 2 year FU, 90% of the patients benefited from the Lateral Menaflex as evidenced by improved clinical outcomes associated with gains in activity and function. Longer-term FU continues to determine the extent and duration of the benefits observed

Purpouse. We hypothesized that patients receiving a medial collagen meniscus implant (MCMI) would show better clinical, radiograpich and Magnetic Resonanace Imaging (MRI) outcomes than patients treated with partial medial meniscectomy (PMM) at minimum 10 year FU. Material and Methods. Thirty-three non-randomized patients (males, mean age 40 years) were enrolled in the study to receive a MCMI (17 patients) or as control treated with a PMM (16 patients). All of them were clinically evaluated at time zero, 5 and minimum 10 years after surgery (mean FU 133 months, range 120–145) by Lysholm, VAS for pain, objective IKDC knee form and Tegner activity level. SF-36 score was performed pre-operatively and at final FU. Bilateral weight-bearing XRays were executed at time zero and at final FU. Minimum 10 years FU MRI images were compared with collected pre-operative MRI images by means of Yulish score. Genovese score was also used to evalute MCMI MRI survivorship. Results. MCMI group showed significantly lower VAS for pain (p = 0.0091), higher objective IKDC (p = 0.0026), Teger index (p = 0.0259) and SF-36 (p = 0.0259 for PHI and p = 0.0036 for MHI) scores compared with PMM group at minimum 10 year FU. Radiographic evaluation showed a significantly lower medial joint line height (p = 0.0002) and side-to-side difference (p = 0.0003) narrowing in MCMI group respect to PMM group at final FU. Discussion. Improvements in pain relief, activity level, objective IKDC score and joint-line preservation are detectable with the use of MCMI at a minimum 10 year FU. On the authors knowledge this is the first long-term controlled trial regarding this device, and our findings confirmed the mid-term good results achieved by Rodkey et al (1). Conclusions. This data support the use of meniscal scaffolds to treat irreparable partial meniscal lesions. Long-term prospective randomized controlled trials on a larger population are necessary to determine the extent and duration of the benefits observed

As arthroplasty demand grows worldwide, the need for a novel cost-effective treatment option for articular cartilage (AC) defects tailored to individual patients has never been greater. 3D bioprinting can deposit patient cells and other biomaterials in user-defined patterns to build tissue constructs from the “bottom-up,” potentially offering a new treatment for AC defects. Novel composite bioinks were created by mixing different ratios of methacrylated alginate (AlgMA) with methacrylated gelatin (GelMA) and collagen. Chondrocytes and mesenchymal stem cells (MSCs) were then encapsulated in the bioinks and 3D bioprinted using a custom-built extrusion bioprinter. UV and double-ionic (BaCl2 and CaCl2) crosslinking was deployed following bioprinting to strengthen bioink stability in culture. Chondrocyte and MSC spheroids were also bioprinted to accelerate cell growth and development of ECM in bioprinted constructs. Excellent viability of chondrocytes and MSCs was seen following bioprinting (>95%) and maintained in culture, with accelerated cell growth seen with inclusion of cell spheroids in bioinks (p<0.05). Bioprinted 10mm diameter constructs maintained shape in culture over 28 days, whilst construct degradation rates and mechanical properties were improved with addition of AlgMA (p<0.05). Composite bioinks were also injected into in vitro osteochondral defects and crosslinked in situ, with maintained cell viability and repair of osteochondral defects seen over a 14-day period. In conclusion, we developed novel composite bioinks that can be triple-crosslinked, facilitating successful chondrocyte and MSC growth in 3D bioprinted scaffolds and in vitro repair of an osteochondral defect model. This offers hope for a new approach to treating AC defects

Objectives. The biomembrane (induced membrane) formed around polymethylmethacrylate (PMMA) spacers has value in clinical applications for bone defect reconstruction. Few studies have evaluated its cellular, molecular or stem cell features. Our objective was to characterise induced membrane morphology, molecular features and osteogenic stem cell characteristics. Methods. Following Institutional Review Board approval, biomembrane specimens were obtained from 12 patient surgeries for management of segmental bony defects (mean patient age 40.7 years, standard deviation 14.4). Biomembranes from nine tibias and three femurs were processed for morphologic, molecular or stem cell analyses. Gene expression was determined using the Affymetrix GeneChip Operating Software (GCOS). Molecular analyses compared biomembrane gene expression patterns with a mineralising osteoblast culture, and gene expression in specimens with longer spacer duration (> 12 weeks) with specimens with shorter durations. Statistical analyses used the unpaired student t-test (two tailed; p < 0.05 was considered significant). Results. Average PMMA spacer in vivo time was 11.9 weeks (six to 18). Trabecular bone was present in 33.3% of the biomembrane specimens; bone presence did not correlate with spacer duration. Biomembrane morphology showed high vascularity and collagen content and positive staining for the key bone forming regulators, bone morphogenetic protein 2 (BMP2) and runt-related transcription factor 2 (RUNX2). Positive differentiation of cultured biomembrane cells for osteogenesis was found in cells from patients with PMMA present for six to 17 weeks. Stem cell differentiation showed greater variability in pluripotency for osteogenic potential (70.0%) compared with chondrogenic or adipogenic potentials (100% and 90.0%, respectively). Significant upregulation of BMP2 and 6, numerous collagens, and bone gla protein was present in biomembrane compared with the cultured cell line. Biomembranes with longer resident PMMA spacer duration (vs those with shorter residence) showed significant upregulation of bone-related, stem cell, and vascular-related genes. Conclusion. The biomembrane technique is gaining favour in the management of complicated bone defects. Novel data on biological mechanisms provide improved understanding of the biomembrane’s osteogenic potential and molecular properties. Cite this article: Dr H. E. Gruber. Osteogenic, stem cell and molecular characterisation of the human induced membrane from extremity bone defects. Bone Joint Res 2016;5:106–115. DOI: 10.1302/2046-3758.54.2000483

Objectives. Bisphosphonates are widely used as first-line treatment for primary and secondary prevention of fragility fractures. Whilst they have proved effective in this role, there is growing concern over their long-term use, with much evidence linking bisphosphonate-related suppression of bone remodelling to an increased risk of atypical subtrochanteric fractures of the femur (AFFs). The objective of this article is to review this evidence, while presenting the current available strategies for the management of AFFs. Methods. We present an evaluation of current literature relating to the pathogenesis and treatment of AFFs in the context of bisphosphonate use. Results. Six broad themes relating to the pathogenesis and management of bisphosphonate-related AFFs are presented. The key themes in fracture pathogenesis are: bone microdamage accumulation; altered bone mineralisation and altered collagen formation. The key themes in fracture management are: medical therapy and surgical therapy. In addition, primary prevention strategies for AFFs are discussed. Conclusions. This article presents current knowledge about the relationship between bisphosphonates and the development of AFFs, and highlights key areas for future research. In particular, studies aimed at identifying at-risk subpopulations and organising surveillance for those on long-term therapy will be crucial in both increasing our understanding of the condition, and improving population outcomes. Cite this article: N. Kharwadkar, B. Mayne, J. E. Lawrence, V. Khanduja. Bisphosphonates and atypical subtrochanteric fractures of the femur. Bone Joint Res 2017;6:144–153. DOI: 10.1302/2046-3758.63.BJR-2016-0125.R1

Introduction. Traumatized musculoskeletal tissue often exhibits prolonged time to healing, mostly due to low blood flow and innervation. Intermittent Pneumatic Compression (IPC) increases blood flow and decreases thromboembolic event after orthopedic surgery,[1] however little is known about healing effects.[2] We hypothesized that IPC could stimulate tissue repair: 1.) blood flow 2.) nerve ingrowth 3.) tissue proliferation and during immobilisation enhance 4.) biomechanical tissue properties. Methods. Study 1: In 104 male Sprague Dawley (SD) rats the right Achilles tendon was ruptured and the animals freely mobilized. Half the group received daily IPC-treatment, using a pump and cuff over the hindpaw that inflates/deflates cyclicly, 0–55mmHg (Biopress SystemTM, Flexcell Int.), and the other half received sham-treatment. Healing was assessed at 1,3,6 weeks by perfusion-analysis with laser doppler scanner (Perimed, Sweden), histology and biomechanical testing. Study 2: 48 male SD-rats were ruptured as above. Three groups of each 16 rats were either mobilized, immobilized or immobilized with IPC treatment. Immobilization was performed by plaster cast. Healing was assessed at 2 weeks with histology and biomechanical testing. Results. Study 1: At 3 and 6 weeks reperfusion increased by 21% and 23% (p< 0.05) after IPC-treatment, strengthened by the observation of elevated numbers of blood vessels and nerves. Fibroblast density was at all time points significantly increased in the IPC group. At three and six weeks the IPC treated tendons displayed an increased tissue organization confirmed by higher collagen I/III ratio in the IPC group. No differences (p = 0.10) were found regarding biomechanical strength. Study 2: Compared to mobilization, immobilization caused a downregulation (p<0.05) of all biomechanical and histological parameters, eg. maximum force decreased 80% and collagen III occurrence by 83%. However when immobilization was combined with IPC biomechanical and histological healing increased significantly compared to pure immobilization, eg. maximum force increased 63% and collagen III occurrence by 150%. Conclusion. This study demonstrated that IPC treatment can counteract biomechanical and morphological deficits caused by immobilization by enhancing proliferative soft tissue repair. Thus, IPC promotes tissue repair by stimulating tissue perfusion and nerve ingrowth as well as accelerating both fibroblast proliferation and collagen organization

Background. Skeletal stem cells (SSCs) have been used for the treatment of osteonecrosis of the femoral head to prevent subsequent collapse. In isolation SSCs do not provide structural support but an innovative case series in Southampton, UK, has used SSCs in combination with impaction bone grafting (IBG) to improve both the biological and mechanical environment and to regenerate new bone at the necrotic site. Aims. Analysis of retrieved tissue-engineered bone as part of ongoing follow-up of this translational case series. Methods. With Proof-of-Concept established in vitro and in vivo, the use of a living bone composite of SSCs and allograft has been translated to four patients (five hips) for treatment of osteonecrosis of their femoral heads. Parallel in vitro culture of the implanted cell-graft construct was performed. Patient follow-up was by serial clinical and radiological examination. In one patient collapse occurred in both hips due to more advanced disease than was originally appreciated. This necessitated bilateral hip arthroplasty, but allowed retrieval of the femoral heads. These were analyzed for Type 1 Collagen production, bone morphology, bone density and mechanical strength by micro computed tomography (CT), histology (A/S stain, Collagen Type 1 immunostain, biorefringence) and mechanical testing. Representative sections of cortical, trabecular and tissue engineered bone were excised from the femoral heads using a diamond-tipped saw-blade and tested to failure by axial compression. Results. Parallel in vitro analysis demonstrated sustained cell growth and viability on the allograft. Three patients currently remain asymptomatic at up to three year follow-up. Histological analysis of the two retrieved femoral heads demonstrated, critically, Type 1 collagen production in the regenerated tissue as well as mature trabecular architecture, indicative of de novo tissue engineered bone. The trabecular morphology of regenerated bone was evident on CT, and this had a bone density of 1400 Grey scale units, (compared to 1200 for natural trabecular bone and 1800 for cortical bone). On axial compressive testing the regenerated bone on the left showed a 24.8% increase in compressive strength compared to ipsilateral normal trabecular bone, and a 22.9% increase on the left. Conclusions. Retrieval analysis data has demonstrated the translational potential of a living bone composite, while ongoing clinical follow-up shows this to be an effective new treatment for osteonecrosis of the femoral head. Regeneration of the necrotic bone may prevent subsequent collapse, thereby delaying, or possibly avoiding, the need for hip arthroplasty in early stage osteonecrosis. Evaluation of this tissue engineering construct has confirmed the potential for clinical treatment of bone defects using SSC based strategies

Introduction. Atypical femoral fracture focused on relation of bisphosphonate use, frequently. However, the mechanism of atypical femoral fracture was not yet clarified. Atypical femoral fractures have been kept femoral shaft cortical thickness and BMD, practically. We hypothesized that atypical femoral fractures were associated with impaired bone quality and curvature of femoral shaft. Materials & Methods. We experienced four atypical femoral fractures. One was subtrochanteric and three were shaft fracture. Two cases received bisphosphonate therapy for 3–5 years. BMD, bone metabolic markers, and bone quality markers were evaluated. Histomorphometry and collagen cross-link analysis were performed. Curvature of femoral shaft and 3-D finite element analysis in one incomplete fracture case were assessed. Results. BMD values were either maintained or not severely decreased. Deterioration of bone quality were verified by the results of histomorphometry, collagen cross-link analysis, and bone quality maker. Especially, homocystine values, such as one of bone quality markers, were increased in all cases. All atypical femoral shaft fractures showed outward curvature of femoral bone. In one case of incomplete atypical femoral shaft fracture, stress was concentrated at the fracture region according 3-D finite element analysis. Conclusions. The results of this study suggest that atypical femoral fractures were estimated associated with deterioration of bone quality and curvature of femoral shaft

Introduction. Bone tissue engineering approaches are an emerging strategy to treat bone defects, and commonly involve the delivery of osteogenic cells and/or drugs via a porous scaffold. We have been exploring an alternative injectable approach for drug delivery that would obviate the need for invasive surgery. Hypothesis. Sucrose Acetate Isobutyrate (SAIB) is a sucrose-based ester that is a highly viscous semi-solid. Diluting SAIB with 10–20% ethanol markedly reduces its viscosity, with ethanol diffusing rapidly after in vivo injection. This phase transitioning property makes SAIB an ideal candidate for bone tissue engineering. Materials and methods. The capacity of SAIB to act as a delivery system for recombinant human BMP-2 (rhBMP-2) was tested in a mouse ectopic bone formation model. In this model SAIB was used to deliver 0 to 10μg rhBMP-2. Next, SAIB was compared with porous collagen scaffold used clinically to delivery rhBMP-2 in a head-to-head trial. Commercial SAIB and SAIB produced in-house were also compared. Bone volumes were quantified by μCT. Discussion. Bone was found to form with as little as 2μg rhBMP-2 when delivered with SAIB. Injected SAIB also showed minimal inflammatory response and rapid breakdown, with bone formation occurring between one and two weeks. Conclusion. SAIB was found to be an effective delivery system for rhBMP-2 with translational utility. Future work will be required to examine the upscaling of this delivery system

Introduction. Nonunions pose complications in fracture management that can be treated using electrical stimulation (ES). Bone marrow mesenchymal stem cells (BMMSCs) are essential in fracture healing, although the effects of different clinical ES waveforms available in clinical practice on BMMSCs cellular activities is unknown. Materials and Methods. We compared Direct Current (DC), Capacitive Coupling (CC), Pulsed Electromagnetic wave (PEMF) and Degenerate Wave (DW) by stimulating human-BMMSCs for 5 days for 3 hours a day. Cytotoxicity, cell proliferation, cell-kinetics and cell apoptosis were evaluated after ES. Migration and invasion were assessed using fluorescence microscopy and affected gene and protein expression were quantified. Results. DW had the greatest proliferative and least apoptotic and cytotoxic effects compared to other waveforms and unstimulated cells after 5 days of ES (p < 0.001). DC, DW and CC resulted in significantly more cells in S phase and G2/M phase (p < 0.01) compared to the unstimulated BMMSCs. CC and DW caused more cells to invade collagen and showed increased MMP-2 and MT1-MMP expression (p < 0.001) compared to the other waveforms and unstimulated BMMSCs. DC increased cellular migration in a scratch-wound assay and all ES waveforms increased migration gene expression with DC having the greatest effect (p < 0.01). Conclusion. The ES waveform is vital in influencing BMMSCs cellular activities. Migration and invasion were increased by ES which suggests that the recruitment of BMMSCs to the healing site during a fracture could be increased by ES

Introduction. Nowadays, autologous platelet-rich plasma is used commonly in wound treatment. However, platelet gel, which was derived from allogeneic platelet-rich plasma (PRP) [1,2], has never been studied about efficacy in vivo or animal models. We aimed to determine efficacy of allogeneic platelet-gel on wound healing in rats by comparing with untreated, antibiotic-gel (Mupirocin 2%) treated and gel (sodium carboxymethylcellulose(NaCMC))-treated control. Methods. Fresh frozen plasma was centrifuged at 1200-G for 15 minutes to extract PRP which would be freeze-dried at −70°c, sterilized with gamma ray of Cobalt source 25 kGy and stored at −70°c. Then, processed freeze-dried PRP was mixed with gel base (NaCMC) as in form of allogeneic platelet-gel concentrated 30 mg/1g by sterilization process (table 1). Full-thickness of 6-mm-diameter skin punch biopsies were performed on 18 female Wistar rats which each rat had four wounds at back. Each wound was applied with untreated care, antibiotic-gel, NaCMC-gel and platelet-gel, respectively. Wound healing was studied from day 0–12. Animals were sacrificed with wound tissues removal on day 3, 7, 12 post-biopsy. Digital planimetric measurement device (VISITRAK, Smith and Nephew) was used in evaluation of total wound area on day 0, 3, 7, 12 post-biopsy. Histopathological changes of wound healing were studied, using 4-μm thickness section with haematoxylin-eosin (H&E) and Masson's trichrome-stain, under light microscope. Results. Platelet-gel reduced wound size more rapidly on day 3, 7 than other groups with statistical significance (p<0.05), although no statistically significant difference compared to antibiotic-treated wounds. Histological study confirmed earlier granulation forming and more collagen fibers in platelet-gel treated group when compared with others. discussion & Conclusions. Allogeneic platelet gel produced the satisfactory efficacy on acute wound healing in rat. This platelet gel needs further study in human for efficacy and safety that might be developed for using in acute wound treatment in the future

Engineered bone tissue to recreate the continuity of damaged skeletal segments is one of the field of interest of tissue engineering. Trabecular titanium has very good mechanical properties and high in vitro and in vivo biocompatibility: it can be used in biomedical applications to promote osteointegration demonstrating that it can be successfully used for regenerative medicine in orthopaedic surgery (1). Purpose of this investigation was to evaluate the behavior of adipose tissue derived stem cells (hASCs) cultured on scaffolds of Trabecular TitaniumTM (Lima-Lto) (TT). hASCs are considered to be multipotent mesenchymal stem cells that are easily induced to differentiate into functional osteoblasts both in vitro and in vivo (2). The hASCs were obtained from the subcutaneous adipose tissue of healthy donors during total hip replacement procedures after digestion with collagenase. They were seeded on monolayer and on the TT scaffolds, and incubated at 37 degrees C in 5% CO2 with osteogenic medium or control medium. The expression of bone-related genes using RT-PCR, time course of alkaline phosphatase activity and morphological investigation with Scanning Electron Microscopy (SEM) were performed to evaluate the osteogenic differentiation of hASCs. Alkaline phosphatase activity, marker of the differentiation toward the osteogenic pattern, was significantly higher in hASCs grown with osteogenic medium than in cells grown with control medium, both in monolayer and TT scaffolds; moreover, also alkaline phosphatase of hASCs grown on TT scaffolds in the presence of control medium increased with time, differently from that of cells grown on monolayer. The osteogenic differentiated hASCs expressed the bone-related genes type I collagen, osteocalcin, Runx-2 and alkaline phosphatase. SEM observations showed that hASCs differentiated toward osteoblast-like cells: they produced a big amount of extracellular matrix that covered the surface of the porous scaffolds with bridges between the pore walls. These data suggest that hASCs are able to adhere to TT scaffolds, to acquire an osteoblastic phenotype and to produce abundant extracellular matrix, with but also without osteogenic medium. We can therefore conclude that this material carries osteinductive properties being responsible of ostegenic differentiation; consequently, this scaffold/cells construct is effective to regenerate damaged tissue and to restore the function of bone tissue

Introduction. Osteoporosis (OP), osteoarthrosis (OA), and rheumatoid arthritis (RA) are the most common age-related degenerative bone diseases, and major public health problems in terms of enormous amount of economic cost. RA is considered as a major cause of secondary osteoporosis. At late stage, OP often leads to skeletal fractures, and OA and RA result in severe joint disability. Over the last a few decades, much significant research on the properties has been carried out on these diseases, however, a detailed comparison of the microarchitecture of cancellous bones of these diseases is not available. In this study, we investigated three-dimensional (3-D) microarchitectural properties of OP, OA and RA cancellous bone. We hypothesized that there were significant differences in microarchitecture among OP, OA and RA bone tissues that might lead to different bone quality. Materials and Method. Twenty OP, fifty OA, and twelve RA femur heads were harvested from patients undergone total hip replacement surgery. Cubic cancellous bone samples (8∗8∗8 mm3) were prepared and scanned with a high resolution microtomographic system (vivaCT 40, Scanco Medical AG., Brüttisellen, Switzerland). Then micro-CT images were segmented using individual thresholds to obtain accurate 3-D data sets. Detailed microarchitectural properties were evaluated based on novel unbiased, model-free 3-D methods. For statistical analysis, one-way ANOVA was used, and a p<0.05 was considered significant. Results. Significant differences in the microarchitecture of cancellous bone were observed among the OP, OA and RA groups. Compared with the other groups, OP cancellous bone had lowest density, thinner, typical rod-like structure and less connectivity (all p<0.01). Interestingly, there were no significant differences in the microarchitectural properties measured between the OA and RA cancellous bones. Both OA and RA cancellous bones had significant higher bone volume fraction and were thicker, typical plate-like structure compared with the OP group (all p<0.01), even though there was clearly bone erosion observed in RA cancellous bone. Discussion. Quantification of the alterations in bone properties and quality will help to gain more insights into the pathogenesis of degenerative bone diseases and to target and develop novel approaches for the intervention and treatment, and for the design, fixation and durability of total joint prosthesis. Our study demonstrated that there were significant differences in the microarchitecture of the OP, OA and RA femur head cancellous bone. The OA and RA cancellous bone had similar bone density and microarchitecture despite apparent bone erosion in the RA cancellous bone. These results from femur head did not support the traditional notion that RA and OP had similar low bone density. Thus, whether femur head bone tissues from these diseases have similar bone collagen, mineral and mechanical properties, more importantly bone quality, should be clarified in the future

Introduction. Dupuytren's disease (DD) is a fibro-proliferative disorder of the palmar fascia whereby a collagen cord contracts affected joints, resulting in flexion deformity that can impair hand function. Currently, surgery is the only effective treatment option in Europe. This 2-part study, consisting of a surgeon survey and chart audit, was designed to assess current surgical practice patterns by DD severity. We report results from the surgeon survey. Methods. A total of 687 participants, including 579 orthopedic surgeons (of which 383 were hand specialists) and 108 plastic surgeons, who had been practicing for >3 and <30 years and operated on 5 DD patients between September and December 2008 were surveyed in 12 countries (UK, Germany, France, Italy, Spain, Hungary, Czech Republic, Poland, Netherlands, Sweden, Denmark, Finland). The survey included queries about procedures performed, factors involved in the decision to use a procedure, satisfaction with the procedure, use of physiotherapy, and recurrence. Results. Regardless of specialty, about 95% of surgeons performed fasciectomy in the previous 12 months. Rates for needle aponeurotomy (NA; 36%) and fasciotomy (70%) were comparable across specialties; a larger proportion of plastic surgeons (65%) used dermofasciectomy (DF) than did orthopaedic (39%) and hand surgeons (44%). Decisions to use NA/fasciotomy were driven mainly by patient comfort and quality-of-life issues (eg, aged >70 y, aesthetics, activity impairment); surgeon satisfaction was linked to shorter recovery times, reduced patient burden, few complications, and restored finger function. Decisions to use open surgeries were based mostly on DD characteristics (eg, contracture severity, speed of progression, recurrence), and surgeon satisfaction was linked to intervention efficacy and durability of the outcome. The percentage of surgeons prescribing physiotherapy and the duration of therapy increased with complexity of the first procedure: NA=86%, 5.3 weeks; fasciotomy=94%, 5.4; fasciectomy=97%, 6.7; and DF=99%, 8.7. On average, 90% of patients were enrolled in a physiotherapy program after undergoing a procedure for DD. Using survey responses, recurrence rates appeared to decrease and time to recurrence increased with procedure complexity: NA=44%, 17 months; fasciotomy=32%, 21; fasciectomy=20%, 29; and DF=20%, 34. Conclusions. To our knowledge, based on the number of participants and countries, this is the largest survey to date to collect, quantify, and describe information about the surgical management of DD in Europe. Although data from all countries were combined and results from the specialties were collapsed, it is a critical first step toward understanding DD treatment patterns. Opportunities to learn more about country- and specialty-specific practices will be presented elsewhere. This study was funded by Pfizer Inc

Introduction. This study reports the results of percutaneous autologous bone marrow grafting in 62 patients with corticosteroids treatment who had one hip osteonecrosis treated with bone marrow (BM) injection and the other contralateral hip osteonecrosis with core decompression (CD) alone. Only patients with bilateral symptomatic osteonecrosis and with those hips at stage I or II (as defined by Steinberg) were included in this study. Material and Methods. Between 1988 and 1995, 62 consecutive patients (28 males and 34 females) were included in this study. These patients had a mean age of 31 years (range 18 to 34 years) at the time of the onset of symptoms. The average follow-up was 17 years (range, 15 to 20 years). An average of 152 + 16 milliliters of marrow was aspirated from the iliac crest. The number of stroma progenitor that was transplanted was estimated by counting the Fibroblast Colony Forming Units which express type I and type III collagen. The bone marrow graft obtained after concentration contained average 4889 + 716 progenitors per cubic centimeter (range 3515 to 6293 per cubic centimeter). Each hip received a mean number of thirty cubic centimeters of bone marrow graft (range 27 to 35 cubic centimeters). The average total number of CFU-F injected in each hip was therefore 147 × 103 cells (range 119 × 103 to 195 × 103 cells). Results. Clinical results were determined by the change in Harris hip scores from preoperative evaluation to the last follow-up visit, by the change in the radiographic progression and by the need of subsequent total hip arthroplasty. Bone marrow grafting afforded better reduction in pain, effected a reduction with time in the number of hips that progressed to collapse, and delayed the need for total hip replacement. Ten hips had collapsed and needed arthroplasty at the most recent follow-up after bone marrow grafting, compared to 45 after core decompression. For hips with collapse, the mean survival time before collapse was 71.2 months (43.35- 60.96; 95% CI) for the bone marrow graft group and 38.5 months for the control group (13.2–39.74; 95% CI). With the number available, there was a positive correlation (Spearman's test) between the duration of clinical survival before collapse and the number or concentration of CFU-F and CFU-GM in the graft in the CD group. These results are explained by the fact that bone marrow injection improved the repair process on MRI and on histology. Overall, 10 hips with bone marrow injection showed a total regression of the signal, 59 hips showed a partial reduction (42 with BM and 17 with CD) and 55 hips did not show a significant reduction (10 with BM and 45 with CD). Conclusion. Bone marrow grafting afforded better reduction in pain, reduction in the number of collapses, delayed the need for total hip replacement, and improved the repair process on MRI and on histology

Aims

Little is known about the effect of haemorrhagic shock and resuscitation on fracture healing. This study used a rabbit model with a femoral osteotomy and fixation to examine this relationship.