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Objective: To determine the accuracy of ultrasound for diagnosing hip luxation in dogs presenting for hind limb lameness., Methods: 24 client-owned dogs presenting with pelvic limb lameness and concern for hip luxation were enrolled in this prospective diagnostic accuracy study from April 1, 2021, to July 1, 2022. An experienced ultrasonographer and a novice ultrasonographer, both masked to the diagnoses, performed hip ultrasonography using a point-of-care ultrasound machine. The experienced evaluator also performed a more comprehensive ultrasonography with a high-end ultrasound machine to characterize concurrent pelvis and hip disease. Pelvic radiographs served as the criterion standard for the diagnosis of hip luxation., Results: Ultrasonographic diagnosis of hip luxation demonstrated high accuracy, ranging from 84.2% to 100% for detecting the presence of luxation and 80.6% to 98.1% for diagnosing the direction of luxation for the novice and experienced evaluators, respectively, with substantial interevaluator agreement (κ = 0.722). Compared to radiographs, ultrasound accurately diagnosed the presence of osteoarthrosis with almost perfect agreement (κ = 0.913) and the presence of fractures of the femoral head/neck and the nonaxial portions of the pelvis with substantial intermodality agreement (κ = 0.775). In 36% of patients, ultrasound identified injury to soft tissue structures that contribute to hip stability, including the gluteal muscles, gemelli, and joint capsule., Conclusions: Ultrasound can be utilized to accurately diagnose the presence of hip luxation and document concurrent hip and pelvic disease., Clinical Relevance: Applications include the use of ultrasonography as part of a routine point-of-care protocol to catalogue injuries in patients with acute trauma and in mobile practice or remote settings where radiography is not readily available.

Cell-derived matrix (CDM) has proven its therapeutic potential and been utilized as a promising resource in tissue regeneration. In this study, we prepared a human fibroblast-derived matrix (FDM) by decellularization of in vitro cultured cells and transformed the FDM into a nano-sized suspended formulation (sFDM) using ultrasonication. The sFDM was then homogeneously mixed with Pluronic F127 and hyaluronic acid (HA), to effectively administer sFDM into target sites. Both sFDM and sFDM containing hydrogel (PH/sFDM) were characterized via immunofluorescence, sol-gel transition, rheological analysis, and biochemical factors array. We found that PH/sFDM hydrogel has biocompatible, mechanically stable, injectable properties and can be easily administered into the external and internal target regions. sFDM itself holds diverse bioactive molecules. Interestingly, sFDM-containing serum-free media helped maintain the metabolic activity of endothelial cells significantly better than those in serum-free condition. PH/sFDM also promoted vascular endothelial growth factor (VEGF) secretion from monocytes in vitro. Moreover, when we evaluated therapeutic effects of PH/sFDM via the murine full-thickness skin wound model, regenerative potential of PH/sFDM was supported by epidermal thickness, significantly more neovessel formation, and enhanced mature collagen deposition. The hindlimb ischemia model also found some therapeutic improvements, as assessed by accelerated blood reperfusion and substantially diminished necrosis and fibrosis in the gastrocnemius and tibialis muscles. Together, based on sFDM holding a strong therapeutic potential, our engineered hydrogel (PH/sFDM) should be a promising candidate in tissue engineering and regenerative medicine.

Background: Degloving injuries represent a challenge in plastic surgery. The aim of this study is to acknowledge the protective effects of hydrogen-rich saline (HRS) solution on a rat hindlimb degloved skin flap., Methods: Twenty-one Sprague-Dawley rats were divided into three groups (control, saline and HRS). Degloving injury model was established, and flaps were sutured back following 5 min of ischemia. The control group did not receive any treatment. The saline group received intraperitoneal physiological saline (10 ml/kg) and the HRS group received intraperitoneal HRS solution (10 ml/kg) postoperatively and daily for 5 days after the operation. Skin samples were obtained for histological, immunohistochemical and biochemical evaluations., Results: Inflammation was lower in the HRS compared with saline (p = 0.02) and control (p = 0.004) groups. Edema was lower in the HRS compared with saline (p = 0.02) and control (p = 0.001) groups. Malondialdehyde (MDA) level was lower in the HRS than the control group (p = 0.01). Total antioxidant level was higher in the HRS compared with saline (p = 0.009) and control (p = 0.03) groups. Total oxidant level was lower in the HRS than the control group (p = 0.02). Oxidative stress index was lower in the HRS compared with saline (p = 0.001) and control (p = 0.0001) groups`. Vascular proliferation was higher in the HRS compared with the control group (p = 0.01)., Conclusion: Repeated HRS injections after trauma increased the viability of skin flap in rat degloving injury model by decreasing local tissue injury, due to its antioxidant, anti-inflammatory and angiogenic effects., Competing Interests: Declaration of Competing Interest Non declared., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Background: The incidence of and mortality due to acute kidney injury is high in patients with traumatic shock. However, it is unclear how hemorrhage and trauma synergistically affect renal function, especially when timely volume resuscitation is not available., Method: We hypothesized that trauma impairs renal tolerance to prolonged hemorrhagic hypotension. Sprague-Dawley rats were randomized into six groups: control, extremity trauma (ET), hemorrhage at 70 mm Hg (70-H), hemorrhage at 55 mm Hg (55-H), ET + 70 mm Hg (70-ETH), and ET + 55 mm Hg (55-ETH). Animals were anesthetized, and ET was induced via soft tissue injury and closed fibula fracture. Hemorrhage was performed via catheters 5 minutes after ET with target mean arterial pressure (MAP) clamped at 70 mm Hg or 55 mm Hg for up to 3 hours. Blood and urine samples were collected to analyze plasma creatinine (Cr), Cr clearance (CCr), renal oxygen delivery (DO2), urinary albumin, and kidney injury molecule-1 (KIM-1)., Results: Extremity trauma alone did not alter renal hemodynamics, DO2, or function. In 70-H, CCr was increased following hemorrhage, while Cr, renal vascular resistance (RVR), KIM-1, and albumin levels remained unchanged. Compared with 70-H, ET + 70 mm Hg exhibited increases in Cr and RVR with decreases in CCr and DO2. In addition, ET decreased the blood volume loss required to maintain MAP = 70 mm Hg by approximately 50%. Hemorrhage at 55 mm Hg and ET + 55 mm Hg exhibited a marked and similar decrease in CCr and increases in RVR, Cr, KIM-1, and albumin. However, ET greatly decreased the blood volume loss required to maintain MAP at 55 mm Hg and led to 50% mortality., Conclusion: These results suggest that ET impairs renal and systemic tolerance to prolonged hemorrhagic hypotension. Thus, traumatic injury should be considered as a critical component of experimental studies investigating outcomes and treatment following hemorrhagic shock., Level of Evidence: This is an original article on basic science and does not require a level of evidence., (Copyright © 2021 Wolters Kluwer Health, Inc. All rights reserved.)

Background: The present study investigated the influence of ischemic postconditioning (I-postC) on the adjustment of renal injury after limb ischemia-reperfusion (I/R) injury, to elucidate the mechanisms of the Toll-like receptor 4 (TLR 4)/NF-κB signaling pathway using histopathological and immunohistochemical methods., Methods: Male Sprague-Dawley rats were randomly assigned to five groups (numbered from 1 to 5): the sham group (Group 1, only the anesthesia procedure was conducted without limb I/R), the I/R group (Group 2, 4 h of reperfusion was conducted following 4 h limb ischemia under anesthesia), the I/R + I-postC group (Group 3, 4 h of ischemia and 4 h of reperfusion was conducted; before perfusion, 5 min of limb ischemia and 5 min of reperfusion were performed in the rats and repeated 3 times), the I/R + TAK group (Group 4, rats were injected with TLR4 antagonist TAK through the caudal vein before limb ischemia and reperfusion under anesthesia), the TAK group (Group 5, rats were injected with TAK, and the anesthesia procedure was conducted without limb I/R). Histological changes in the kidney in different groups were observed, and the extent of tubular injury was assessed. Changes in biochemical indexes and the expression of inflammatory factors, TLR4, and NF-κB were also evaluated., Results: Compared with rats in the I/R group, the secretion of inflammatory factors and the expression levels of TLR4 and NF-κB were decreased in rats in the I/R + I-postC group. Histological analysis revealed renal injury, including inflammatory cell infiltration, dilatation of the tubuli lumen, congestion in glomerular capillaries, degeneration of tubuli epithelial cells, and necrosis was ameliorated by I-postC. Immunohistochemical studies showed that I/R-induced elevation in TLR4 and NF-κB expression was reduced by I-postC treatment. Moreover, the expression levels of TLR4, NF-κB, and inflammatory factors in rats in the I/R + TAK group were also decreased, and the renal pathological lesion was alleviated, which was similar to that in rats in the I/R + I-postC group., Conclusions: The present findings suggest that I-postC can reduce tissue injury and kidney inflammation induced by limb I/R injury, possibly via inhibition of the TLR4 and NF-κB pathways.

Pedal bone fractures are one of the most common fracture locations in adult cattle and can be diagnosed by radiographs in two planes. Most bovine practitioners do not have access to such X-ray machines, but many use ultrasound units on a daily basis, primarily for reproductive medicine. For this reason, in this double-masked, randomized controlled study, we aimed to investigate the suitability of ultrasonographic examination using a 5 MHz linear transducer for diagnosing closed fractures of the pedal bone in cattle. A total of 54 hindlimb claws from slaughtered cattle were prepared and approximately 50% of the claws were artificially fractured. All claws were ultrasonographically examined twice by two examiners to determine the presence or absence of fractures and their locations. Ultrasound results were confirmed using radiographs of the claws as the reference standard method. All fracture locations as determined by ultrasonography were situated within ±2 mm of the radiographically-determined fracture zone. Ultrasound examination yielded a calculated sensitivity of 93%, a specificity of 91% and an inter-rater reliability of 0.77. The intra-rater reliability for the examiners were 0.96 and 0.88. Examiner experience with ultrasound examination and using ultrasound images for diagnosis could have influenced diagnostic accuracy. We conclude that artificially-created pedal bone fractures in ex-vivo bovine claws can be diagnosed using ultrasonography; similar results are expected in live animals. These results should encourage veterinarians to use ultrasonography for diagnosing pedal bone fractures in cattle., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Background: Postinjury hypercoagulability occurs in >25% of injured patients, increasing risk of thromboembolic complications despite chemoprophylaxis. However, few clinically relevant animal models of posttraumatic hypercoagulability exist. We aimed to evaluate a rodent model of bilateral hindlimb injury as a preclinical model of postinjury hypercoagulability., Methods: Forty Wistar rats were anesthetized with isoflurane: 20 underwent bilateral hindlimb fibula fracture, soft tissue and muscular crush injury, and bone homogenate injection intended to mimic the physiological severity of bilateral femur fracture. Twenty sham rats underwent anesthesia only. Terminal citrated blood samples were drawn at 0, 6, 12, and 24 hours (n = 5 per timed group) for analysis by native thromboelastography in the presence and absence of taurocholic acid to augment fibrinolysis. Plasminogen activator inhibitor 1 and α-2 antiplasmin levels in plasma were assessed via enzyme-linked immunosorbent assay., Results: Injured rats became hypercoagulable relative to baseline by 6 hours based on thromboelastography maximal amplitude (MA) and G (p < 0.005); sham rats became hypercoagulable to a lesser degree by 24 hours (p < 0.005). Compared with sham animals, injured rats were hypercoagulable by MA and G within 6 hours of injury, remained hypercoagulable by MA and G through at least 24 hours (all p < 0.01), and showed impaired fibrinolysis by taurocholic acid LY30 at 12 hours (p = 0.019) and native LY30 at 24 hours (p = 0.045). In terms of antifibrinolytic mediators, α-2 antiplasmin was elevated in trauma animals at 24 hours (p = 0.009), and plasminogen activator inhibitor 1 was elevated in trauma animals at 6 hours (p = 0.004) and 12 hours (p < 0.001) when compared with sham., Conclusions: Orthopedic injury in rodents induced platelet and overall hypercoagulability within 6 hours and fibrinolytic impairment by 12 to 24 hours, mimicking postinjury hypercoagulability in injured patients. This rodent model of orthopedic injury may serve as a preclinical testing ground for potential therapies to mitigate hypercoagulability, maintain normal fibrinolysis, and prevent thromboembolic complications.

An adult red-legged seriema ( Cariama cristata ) presented with a comminuted fracture of the tibiotarsus and fibula. Surgery was performed, and a type II external fixator, with 2 distal and 2 proximal pins, was used to stabilize the fracture. After a 10-day stabilization period, the bird developed a second fracture on the same bone, proximal to the first fracture site. Another surgery was performed on the seriema similar to the first one. However, in this second surgical procedure a single pin, instead of 2 perpendicular pins, was placed proximally to the fracture site. After the second surgical procedure, bone marrow stem cells (BMSCs) from the seriema's left ulna were collected. Twenty-seven days after the second surgery, the BMSCs were transplanted, into the fracture sites. Twenty-four days after the stem cells were injected into the fractures (51 days after the second surgical procedure), radiographic images revealed healing bone calluses at the fracture sites. The fracture healing was relatively long for this case (a total of 75 days). The addition of bone marrow stem cell therapy to the use of external fixation may have contributed to the healing observed radiographically 24 days after administration; therefore, bone marrow stem cell therapy, in addition to traditional surgical fracture reduction and stabilization, may be a promising therapeutic approach for avian cases with similar injuries and bone anatomy. However, as this is a single case, this therapeutic modality deserves further application and study. Moreover, we suggest modifications in the bone marrow stem cell collection and therapy, which may be useful for future studies and application involving birds.

A 2-month-old goat and a 5-year-old sheep were referred to the clinic due to severe lameness. The goat had a 5-week-old infected comminuted, left metacarpal fracture with extended osteomyelitis of the distal metacarpal bones. The sheep presented with an open oblique fracture of the left tibia and a concurrent, perforating wound of the fetlock joint in the same hindlimb. In both cases, limb amputation was performed at the level of the shoulder joint and the proximal femur, respectively, under general anesthesia combined with blocks of the large proximal limb nerves. The surgical procedure was successful in both cases and the animals showed good postoperative mobility and quality of life. Limb amputations in small ruminants are rarely described, but may offer a valuable salvage procedure instead of euthanasia., Competing Interests: Die Autoren bestätigen, dass kein Interessenkonflikt besteht., (© Georg Thieme Verlag KG Stuttgart · New York.)

Neonatal tissue damage induces long-term deficits in inhibitory synaptic transmission within the spinal superficial dorsal horn (SDH) that include a reduction in primary afferent-evoked, feedforward inhibition onto adult projection neurons. However, the subpopulations of mature GABAergic interneurons which are compromised by early-life injury have yet to be identified. The present research illuminates the persistent effects of neonatal surgical injury on the function of inhibitory SDH interneurons derived from the prodynorphin (DYN) lineage, a population that synapses directly onto lamina I spinoparabrachial neurons and is known to suppress mechanical pain and itch in adults. The results demonstrate that hindpaw incision at postnatal day 3 (P3) significantly decreased the strength of primary afferent-evoked glutamatergic drive onto DYN neurons within the adult mouse SDH while increasing the appearance of afferent-evoked inhibition onto the same population. Neonatal injury also dampened the intrinsic membrane excitability of mature DYN neurons, and reduced their action potential discharge in response to sensory input, compared with naive littermate controls. Furthermore, P3 incision decreased the efficacy of inhibitory DYN synapses onto adult spinoparabrachial neurons, which reflected a prolonged reduction in the probability of GABA release. Collectively, the data suggest that early-life tissue damage may persistently constrain the ability of spinal DYN interneurons to limit ascending nociceptive transmission to the adult brain. This is predicted to contribute to the loss of feedforward inhibition onto mature projection neurons, and the "priming" of nociceptive circuits in the developing spinal cord, following injuries during the neonatal period. SIGNIFICANCE STATEMENT Neonatal injury has lasting effects on pain processing in the adult CNS, including a reduction in feedforward inhibition onto ascending projection neurons in the spinal dorsal horn. While it is clear that spinal GABAergic interneurons are comprised of multiple subpopulations that play distinct roles in somatosensation, the identity of those interneurons which are compromised by tissue damage during early life remains unknown. Here we document persistent deficits in spinal inhibitory circuits involving dynorphin-lineage interneurons previously implicated in gating mechanical pain and itch. Notably, neonatal injury reduced the strength of dynorphin-lineage inhibitory synapses onto mature lamina I spinoparabrachial neurons, a major output of the spinal nociceptive network, which could contribute to the priming of pain pathways by early tissue damage., (Copyright © 2020 the authors.)

Mesenchymal stem cells (MSCs) encapsulation by three-dimensionally (3D) printed matrices were believed to provide a biomimetic microenvironment to drive differentiation into tissue-specific progeny, which made them a great therapeutic potential for regenerative medicine. Despite this potential, the underlying mechanisms of controlling cell fate in 3D microenvironments remained relatively unexplored. Here, we bioprinted a sweat gland (SG)-like matrix to direct the conversion of MSC into functional SGs and facilitated SGs recovery in mice. By extracellular matrix differential protein expression analysis, we identified that CTHRC1 was a critical biochemical regulator for SG specification. Our findings showed that Hmox1 could respond to the 3D structure activation and also be involved in MSC differentiation. Using inhibition and activation assay, CTHRC1 and Hmox1 synergistically boosted SG gene expression profile. Together, these findings indicated that biochemical and structural cues served as two critical impacts of 3D-printed matrix on MSC fate decision into the glandular lineage and functional SG recovery., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).)

Objectives: Conditional Alk2 Q207D-floxed ( caALK2 fl ) mice have previously been used as a model of heterotopic ossification (HO). However, HO formation in this model can be highly variable, and it is unclear which methods reliably induce HO. Hence, these studies report validated methods for reproducibly inducing HO in caALK2 fl mice., Methods: Varying doses of Adex-cre and cardiotoxin (CTX) were injected into the calf muscles of 9, 14, or 28-day-old caALK2 fl/- or caALK2 fl/fl mice. HO was measured by planar radiography or microCT at 14-28 days post-injury., Results: In 9-day-old caALK2 fl/- or caALK2 fl/fl mice, single injections of 10 9 PFU Adex-cre and 0.3 μg of CTX were sufficient to induce extensive HO within 14 days post-injury. In 28-day-old mice, the doses were increased to 5 x 10 9 PFU Adex-cre and 3.0 μg of CTX to achieve similar consistency, but at a slower rate versus younger mice. Using a crush injury, instead of CTX, also provided consistent induction of HO. Finally, the Type 1 BMPR inhibitor, DMH1, significantly reduced HO formation in 28-day-old caALK2 fl/fl mice., Conclusions: These data illustrate multiple methods for reliable induction of localized HO in the caALK2 fl mouse that can serve as a starting point for new laboratories utilizing this model., Competing Interests: Dr. Charles Hong reports grants, personal fees and other from La Jolla Pharmaceutical Company, during the conduct of the study; In addition, Dr. Charles Hong has a patent Fused heterocyclic compounds as selective BMP inhibitors licensed to La Jolla Pharmaceutical Co. Yuji Mishina receives research grant money from NIH and AMED (a funding agency in Japan). Dr. Daniel Perrien reports grants from NIH, grants from the Department of Veterans Affairs during the conduct of the study. Haichun Pan and Nicole Fleming have nothing to disclose.

Objectives: This study aims to investigate the changes in bone morphogenetic protein-2 (BMP-2) expression and mechanical properties in the healing process of rats with osteoporotic hindlimb fracture., Methods: 120 rat models of osteoporotic hindlimb fracture were established and randomly divided into experimental group and control group. Quantitative real-time polymerase chain reaction (PCR) used to detect the BMP-2 expression in the rat's callus tissue on the fractured side. The mechanical properties of rat's hindlimb skeleton were examined using a universal material mechanics testing machine., Results: The BMP-2 expression in the experimental group was higher than that in the control group (p<0.05). The linear correlation analysis showed that the BMP-2 was positively correlated with healing time (r=0.87, p<0.05). The mechanical properties were markedly improved at T2, T3 and T4, which peaked at T4 (p<0.05). However, the mechanical properties in the rats in the experimental group were notably superior to those in the control group at T2, T3, and T4 (p<0.05)., Conclusions: The treatment with strontium ranelate can effectively improve the BMP-2 and bone mechanical properties of the rats with osteoporotic hindlimb fracture in the healing stage and accelerate the healing progress, which could be proved to be an efficacious means in treating osteoporotic fracture., Competing Interests: The authors have no conflict of interest.

Introduction: Commercially available junctional tourniquets (JTQs) have several drawbacks. We developed a low-cost, compact, easy to apply JTQ. The aim of this study was to assess the tourniquets' safety and efficacy in a swine model of controlled hemorrhage., Materials and Methods: Five pigs were subjected to controlled bleeding of 35% of their blood volume. Subsequently, the JTQ was applied to the inguinal area for 180 minutes. Afterwards, the tourniquet was removed for additional 60 minutes of follow up. During the study, blood flow to both hind limbs and blood samples for tissue damage markers were repeatedly assessed. Following sacrifice, injury to both inguinal areas was evaluated microscopically and macroscopically., Results: Angiography demonstrated complete occlusion of femoral artery flow, which was restored following removal of the tourniquet. No gross signs of tissue damage were noticed. Histological analysis revealed mild necrosis and infiltration of inflammatory cells. Blood tests showed a mild increase in potassium and lactic acid levels throughout the protocol., Conclusions: The tourniquet achieved effective arterial occlusion with minimal tissue damage, similar to reports of other JTQs. Subjected to further human trials, the tourniquet might be a suitable candidate for widespread frontline deployment because of its versatility, compactness, and affordable design., (© Association of Military Surgeons of the United States 2020. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

INTRODUCTION Vitamin D-deficiency is known to cause nerve conduction impairments, cancer and chronic diseases, as well as the pathogenesis of osteoarthritis. Our goal with this study is to evaluate the cartilage healing by applying intraarticular 1α, 25 (OH) 2D3 at different doses in rats with normal vitamin D levels and metabolism, which we made focal chondral damage model in the knee joint. MATERIAL AND METHODS 35 male Sprague-Dawley rats aged 20-24 weeks were used in our study. Both knees of rats were cartilage defected surgically on day 0. Joint injections performed at 06:00 am on 0th and 2nd days and after second injection others performed on days 9-16 and 23 following a weekly period. RESULTS In the fourth week, hematoxylin eosin staining measurements showed statistically significant difference according to the groups (p < 0.01) Metalloproteinase-13 (MMP-13) in histological staining for evaluating cartilage healing and healing levels showed statistically significant differences between the groups at first week and fourth week (p < 0.05). DISCUSSION Vitamin D, which affects many tissues through its receptors, is believed to be chondroprotective and neuroprotective by decreasing the expression of MMP in cartilage fibroblast, macrophage, lymphocyte through its intracellular receptors. To the best of our knowledge, this is the first study known to be intraarticular use of 1α, 25-dihydroxyvitamin D3. Our study has been found to be safe and successful in terms of weight, systemic PTH and 1α, 25-dihydroxyvitamin D3 levels in rats during treatment as well as better healing of cartilage damage. Key words: vitamin D3 receptor, articular cartilage, orthopedics, nerve conduction.

Generating universal human umbilical mesenchymal stem cells (UMSCs) without immune rejection is desirable for clinical application. Here we developed an innovative strategy using CRISPR/Cas9 to generate B2M - UMSCs in which human leucocyte antigen (HLA) light chain β2-microglobulin (B2M) was deleted. The therapeutic potential of B2M - UMSCs was examined in a mouse ischaemic hindlimb model. We show that B2M - UMSCs facilitated perfusion recovery and enhanced running capability, without inducing immune rejection. The beneficial effect was mediated by exosomes. Mechanistically, microRNA (miR) sequencing identified miR-24 as a major component of the exosomes originating from B2M - UMSCs. We identified Bim as a potential target of miR-24 through bioinformatics analysis, which was further confirmed by loss-of-function and gain-of-function approaches. Taken together, our data revealed that knockout of B2M is a convenient and efficient strategy to prevent UMSCs-induced immune rejection, and it provides a universal clinical-scale cell source for tissue repair and regeneration without the need for HLA matching in the future., (© 2019 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.)

Fractures of the tibia and fibula are common in dogs and cats and occur most commonly as a result of substantial trauma. Tibial fractures are particularly amenable to treatment using minimally invasive fracture repair (MIFR) techniques that preserve blood supply to comminuted fracture fragments, accelerating bone callus production and speeding fracture healing. Treatment of tibial fractures using MIFR techniques has been found to reduce surgical time, reduce the time for fracture healing, and to decrease patient morbidity, while at the same time reducing complications compared with traditional open reduction and internal fixation., (Copyright © 2019 Elsevier Inc. All rights reserved.)

A thorough working knowledge of the anatomic landmarks of the femur facilitates anatomic alignment during minimally invasive osteosynthesis (MIO). A variety of fixation techniques, including plate, plate-rod, and interlocking nail, are well suited for stabilization of femoral shaft fractures with MIO techniques. Axis and torsional alignment can be assessed with various intraoperative techniques to ensure that anatomic alignment is obtained., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Introduction: Extremity tourniquet (TNK) application is an effective means of achieving compressible hemorrhage control in the emergency prehospital and clinical trauma setting. Modern United States military medical doctrine recommends TNK use to prevent lethal hemorrhage from extremity injury, followed by systemic prophylactic antibiotics to prevent wound infection. Because tissue pharmacokinetics of prophylactic antimicrobials during and after TNK-induced limb ischemia are largely unknown, this study was conducted to empirically determine the relationship between TNK application time and soft tissue antibiotic exposure in order to guide medical personnel in the management of extremity trauma., Materials and Methods: Hind limbs of anesthetized male Sprague Dawley rats were exsanguinated, and ischemia maintained by a pneumatic cuff placed at the level of the mid femur on one limb; the non-ischemic contralateral limb served as comparison tissue. Systemic prophylactic antibiotics (cefazolin, moxifloxacin, or ertapenem) were administered intravenously before or after TNK release following 2 or 4 h of ischemia with subsequent re-dosing every 12 h for 3 days. Free antibiotic in the interstitial fluid (ISF) of the tibialis anterior muscle of both hind limbs was recovered via microdialysis during ischemia and over three periods during reperfusion: immediately following TNK release, at 24 h post TNK release, and at 72 h post TNK release. Plasma and ISF free drug concentrations were determined by high-performance liquid chromatography., Results: Tourniquet application prevented delivery of prophylactic antibiotics into distal soft tissue for the duration of ischemia, and caused a profound reduction in skeletal muscle drug exposure for up to 72 h following TNK release. A progressive decline in tissue antibiotic exposure during reperfusion was observed as TNK times increased from 2 h to 4 h. The timing and severity of reduced drug distribution in post-ischemic skeletal muscle varied substantially among the three antibiotic classes evaluated., Conclusions: Prolonged tourniquet application can significantly reduce distribution of prophylactic antibiotics into soft tissue during and after ischemia, potentially impairing prophylaxis of extremity wound infection. Our findings support the examination of alternative approaches to wound infection prophylaxis under conditions of delayed casualty evacuation when occlusive hemorrhage control measures are utilized., (Copyright © 2019. Published by Elsevier Ltd.)

Injured tendons heal through the formation of a fibrovascular scar that has inferior mechanical properties compared to native tendon tissue. Reducing inflammation that occurs as a result of the injury could limit scar formation and improve functional recovery of tendons. Prostaglandin D 2 (PGD 2 ) plays an important role in promoting inflammation in some injury responses and chronic disease processes, and the inhibition of PGD 2 has improved healing and reduced disease burden in animal models and early clinical trials. Based on these findings, we sought to determine the role of PGD 2 signaling in the healing of injured tendon tissue. We tested the hypothesis that a potent and specific inhibitor of hematopoietic PGD synthase (HPGDS), GSK2894631A, would improve the recovery of tendons of adult male rats following an acute tenotomy and repair. To test this hypothesis, we performed a full-thickness plantaris tendon tenotomy followed by immediate repair and treated rats twice daily with either 0, 2, or 6 mg/kg of GSK2894631A. Tendons were collected either 7 or 21 days after surgical repair, and mechanical properties of tendons were assessed along with RNA sequencing and histology. While there were some differences in gene expression across groups, the targeted inhibition of HPGDS did not impact the functional repair of tendons after injury, as HPGDS expression was surprisingly low in injured tendons. These results indicate that PGD 2 signaling does not appear to be important in modulating the repair of injured tendon tissue., (© 2019 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society.)

Background: Hypothermic ex situ perfusion (HESP) systems are used to prolong allograft survival in solid organ transplantations and have been shown to be superior to static cold storage (SCS) methods. However, the effect of this preservation method on limb allograft survival and long-term function has not yet been tested. In this study, we investigated the long-term effects of the HESP on skeletal muscle metabolism, structure, and force generation and compared it with the current standard of preservation., Methods: Forty male Lewis rats (250 ± 25 g) were divided into 5 groups, including naive control, sciatic nerve transection or repair, immediate transplantation, SCS, and HESP. For the SCS group, limbs were preserved at 4°C for 6 hours. In the HESP group, limbs were continuously perfused with oxygenated histidine-tryptophan-ketoglutarate (HTK) solution at 10-15°C for 6 hours. Hemodynamic and biochemical parameters of perfusion were recorded throughout the experiment. At 12 weeks, electromyography and muscle force measurements (maximum twitch and tetanic forces) were obtained along with muscle samples for histology and metabolomics analysis., Results: Histology demonstrated 48% myocyte injury in the HESP group compared with 49% in immediate transplantation (P = 0.96) and 74% in the SCS groups (P < 0.05). The maximum twitch force measurement revealed a significantly higher force in the HESP group compared with the SCS group (P = 0.029). Essential amino acid levels of the gastrocnemius muscle did not reach significance, with the exception of higher proline levels in the HESP group., Conclusions: HESP using HTK protects viability of the limb but fails to restore muscle force in the long term.

The modern dairy industry is plagued by a high prevalence of claw horn lesions in cows, which cause lameness, affect well-being, limit milk production, and are responsible for premature removal of cows from the herd. The lateral hind claws are primarily affected, and this has been linked to a relatively higher load being exerted on the lateral claws when cows shift weight from one hind limb to the other. The vertical ground reaction forces and mean and maximum pressures under the claws were measured in 40 nonlame dairy cows before and during a shift in weight from one hind limb to the other, which was accomplished by applying pressure manually to one side of the pelvis. During square standing on firm ground, about two-thirds of the entire hind limb load was exerted on the 2 lateral claws, and the remaining one-third was exerted on the medial claws combined. At the moment of maximum weight shift, the lateral claw of the loaded limb bore almost two-thirds of the entire load of both hind limbs, with the heel zone bearing almost half of the load of both hind limbs. Subsequently, the load of the lateral claw of the contralateral hind limb decreased, as did the load of both medial claws. Thus, the weight redistribution had occurred predominantly between the lateral hind claws. The high load exerted on a lateral hind claw during weight shift and at maximum weight shift is assumed to play a role in the pathogenesis of claw horn lesions, particularly when accentuated by a softened claw horn and hard flooring., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The use of mouse models as a tool to study ankle sprain requires a basic understanding of the similarities and differences between human and mouse ankle joint anatomy. However, few studies have been conducted that address the merits and drawbacks of these differences in the functioning of joints. Twenty hindfoot specimens were obtained from 10 male C57BL/6J mice and scanned using micro-CT. The foot and ankle skeletal structures were reconstructed in three dimensions. Morphological parameters were then measured using a plane projection method and normalized data were compared with those of human ankles. There was no significant difference in the malleolar width, maximal tibial thickness, tibial arc length, trochlea tali arc length or trochlea tali width of the mouse specimens compared with the human model. However, a groove was observed on the talar dome in the mouse specimens which was not observed in humans, the talar dome being more symmetric. The mouse ankle was to a large extent able to mimic the mechanism of a human ankle and so a mouse model could be appropriate for expanding our understanding of ankle biomechanics in general. However, the structural differences in the talar dome in the mouse and human should not be ignored. Although there are some differences in the mouse and human ankle that cannot be ignored, compared to other animals, the human ankle is more similar to that of the mouse.

Anterior cruciate ligament (ACL) and meniscus injuries are highly correlated with post-traumatic knee osteoarthritis (PTOA). The aim of this study was to examine whether early intervention with low-intensity pulsed ultrasound (LIPUS) at an intensity of 0.1 W/cm 2 helps delay PTOA progression. A PTOA model was established by ACL transection and meniscectomy in male Sprague-Dawley rats. LIPUS intervention (1.0 MHz, 0.1 W/cm 2 ) started on the third day after surgery and continued for 4 consecutive wk. Histopathological analyses and immunoassays of collagen type II and matrix metallopeptidase 13 in joints were conducted. Results indicated that compared with the sham treatment, LIPUS significantly reduced Mankin scores, inflammatory cells and matrix metallopeptidase 13 expression and increased collagen type II expression in rats with PTOA (p < 0.05). Early intervention with LIPUS has beneficial effects on delaying cartilage degradation by reducing synovial inflammation and matrix metallopeptidase 13 expression, as well as enhancing collagen type II expression in cartilage., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Background: The goal of this study was to characterize the properties of human CD34 + cells in culture and investigate the feasibility and efficacy of CD34 + transplantation in a mouse model of limb ischemia and in patients with no-option critical limb ischemia., Methods: Human CD34 + cells isolated from peripheral blood and grown in culture for up to four passages stained positively for the surface markers CD34 and CD133 and showed high viability after cryopreservation and recovery. Seven days after surgery to induce limb ischemia, ischemic muscles of nude mice were injected with CD34 + cells. Two weeks later, mice were scored for extent of ischemic injury, and muscle tissue was collected for immunohistochemical analysis of vascular endothelial cells and RT-PCR analysis of cytokine expression., Results: Injury scores of CD34 + -treated, but not control, mice were significantly different before and after transplantation. Vascular density and expression of VEGF and bFGF mRNAs were also significantly increased in the treated mice. Patients with severe lower extremity arterial ischemia were injected with their own CD34 + cells in the affected calf, foot, or toe. Significant improvements were observed in peak pain-free walking time, ankle-brachial index, and transcutaneous partial oxygen pressure. These findings demonstrate that growth of human CD34 + cells in vitro and cryopreservations are feasible., Conclusion: Such cells may provide a renewable source of stem cells for transplantation, which appears to be a feasible, safe, and effective treatment for patients with critical limb ischemia., (© 2018 Wiley Periodicals, Inc.)

Background:  Targeted muscle reinnervation (TMR) is a novel approach to postamputation neuroma pain; however, this has not been explicitly studied. The purpose of this study was to develop a TMR model in hind limb amputated rats., Methods:  Ten hind limbs from 5 Sprague Dawley cadaver rats were used. Sciatic nerve, main branches of the sciatic nerve (common peroneal, tibial, sural), motor branches from the sciatic nerve to the biceps femoris and cauda femoris, gluteal nerve and its motor branches to the semimembranosus, and biceps femoris and femoral nerve were dissected to look for consistent nerve anatomy that can be used for TMR in the rat hind limb amputation model. Transfemoral amputation was performed and two types of coaptations were made: common peroneal nerve to motor branch to biceps femoris and tibial nerve to motor branch to semimembranosus., Results:  The total surgical time for the dissection, amputation, and coaptation of nerves was ∼90 minutes. A total of 100 nerves were dissected in 10 rat hind limbs. Anatomical dissections were straightforward to perform. Anatomy of the dissected nerves was consistent. Hind limb amputations were performed without damaging the target muscles and nerves. Nerve lengths were sufficient for coaptation without any tension., Conclusions:  To the best of our knowledge, this is the first report on TMR model in hind limb amputated rats. This model will allow for mechanical, electromyography (EMG), and histological analysis for future assessment of neuroma prevention., Competing Interests: None., (Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.)

Tibiotarsal fractures are a common presentation in small bird species and anecdotally have been reported to carry a good prognosis with proper treatment, such as external coaptation. For this retrospective study, the medical records of 5 institutions were reviewed for tibiotarsal fractures diagnosed in companion birds weighing less than 200 g. A total of 86 cases met the inclusion criteria. Cockatiels ( Nymphicus hollandicus) (24/86) and budgerigars ( Melopsittacus undulatus) (19/86) were the most frequently represented species. Median body weight of the birds included was 72 g (range, 16-182 g). Mid-diaphyseal (46/86) and closed (73/86) fractures with intact, deep pain sensation in the affected limb (69/76) were most frequent. A tape splint alone (79/86) or a tape splint in addition to an intramedullary pin (7/86) were applied in all cases. Median time to fracture stabilization based on palpation was 19 days (range, 7-49 days). In most cases (61/86), the initial splint applied was maintained until fracture healing was complete. A successful outcome was documented in 92% (79/86) of birds. Fractures caused by a dog or cat attack, birds presenting without deep pain sensation in the affected limb, and cases where the splint was removed before 14 days after fixation were associated with a significantly increased risk of complications, resulting in an unsuccessful outcome. The findings of this study indicated that a tape splint is an appropriate means for treatment of tibiotarsal fractures in birds weighing less than 200 g.

Purpose: Recombinant human hyaluronidase (rHuPH20) is widely used as a spreading factor, which enhances the absorption of subcutaneously injected medicines. The anti-inflammatory and anti-edema effects of the enzyme were demonstrated in previous studies. In the present study, the anti-edema effect of rHuPH20 was compared with that of dexamethasone in a traumatic rat paw edema model., Materials and Methods: Twenty-four Sprague-Dawley rats (weight 200 to 450 g) were divided into 3 groups: control (group 1), rHuPH20 (group 2), and dexamethasone (group 3). Traumatic edema was induced in the right hind paws of the rats using Feeney's weight-drop model. After edema induction, 0.4 mL of rHuPH20 (100 U/kg = 0.88 μg/kg dose) and 0.4 mL of dexamethasone (0.5 mg/kg dose) were injected into the right hind paws of the rats in groups 2 and 3. The paw volumes were measured before edema induction and at 3, 6, 12, 24, 48, and 72 hours after induction using a plethysmometer. The Mann-Whitney U test was used for the statistical analyses. Probabilities < .05 were accepted as statistically significant., Results: The between percentage change in the edema mean values of groups 1 and 3 showed no significant difference at all time points; however, group 2 showed significantly less change in the edema mean values at 3, 6, 12, 24, and 48 hours after edema induction (P < .05) compared with group 1. The change in the edema mean value for group 2 was significantly less than that for group 3 at 3, 6, 12, 24, and 48 hours after edema induction (P < .05)., Conclusions: Local rHuPH20 injection more effectively reduced the edema that was induced traumatically in rat paws than did dexamethasone. However, further clinical studies are needed regarding the use of rHuPH20 as a postoperative anti-edema agent in place of dexamethasone., (Copyright © 2018 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.)

Background: Pain management is important in prehospital care of patients with extremity trauma (ET). The goal of this study was to establish a rat model of ET for prehospital pain research and validate it using pain behaviors and analgesics., Methods: Rats were anesthetized using isoflurane, and ET was induced in one hindlimb via clamping retrofemoral tissues for 30 seconds, followed by closed fibula fracture. Rats regained consciousness after ET. Pain responses in the injured hindlimb to thermal hyperalgesia (paw withdrawal latency [PWL]), mechanical allodynia (paw withdrawal pressure [PWP]), and weight bearing (WB) were determined before and 90 minutes after ET. Morphine (2 mg/kg), fentanyl (10 μg/kg), sufentanil (1 μg/kg), ketamine (5 mg/kg), or vehicle (saline) were then administered via intravenous (i.v.) injection, followed by PWL, PWP, and WB assessments at 10 minutes, 40 minutes, 80 minutes, and 120 minutes after analgesia., Results: After ET, PWL, PWP, and WB were significantly decreased by 61 ± 4%, 64 ± 8%, and 65 ± 4%, respectively, compared with pre-ET values. These pain behaviors were maintained for 3 hours to 4 hours. Compared with the saline group, opioid analgesics significantly increased PWL for at least 80 minutes, with sufentanil exhibiting the highest analgesic effect. An increase in PWL was only observed at 10 minutes after ketamine. The PWP was transiently increased with opioid analgesics for 10 minutes to 40 minutes, but was not changed with ketamine. Weight bearing was improved with opioid analgesics for at least 2 hours, but only for up to 80 minutes with ketamine., Conclusion: Our ET model includes long bone fracture and soft tissue injury, but no fixation surgery, mimicking prehospital ET. Our model produces acute, steady, and reproducible trauma-related pain behaviors, and is clinically relevant regarding the pain behaviors and established responses to common analgesics. This model of acute pain due to ET is ideal for prehospital pain management research.

The purpose of this retrospective study was to report the outcomes of 19 dogs and 1 cat undergoing reverse saphenous conduit flap between 1999 and 2016. Reverse saphenous conduit flap was used to treat traumatic wounds and wounds resulting from tumor excision in the hind limb; the majority of cases had medial shearing injuries. All animals had complete flap survival. In five animals (20%), minor donor site dehiscence occurred, which did not require surgery. Other postoperative complications included signs of severe venous congestion in one dog. Reverse saphenous conduit flap is a useful technique to repair skin defects of the distal hind limb.

The purpose of this study is to determine the feasibility of using murine models for translational study of knee ligament injury, repair, and reconstruction. To achieve this aim, we provide objective, quantitative data detailing the gross anatomy, biomechanical characteristics, and microscopic structure of knee ligaments of 44 male mice (C57BL6, 12 weeks of age). Biomechanical testing determined the load-to-failure force, stiffness, and the site of ligament failure for the anterior cruciate ligament (ACL), posterior cruciate ligament (PCL), and the medial and lateral collateral ligaments (MCL and LCL). These data are complemented by histological characterization of each of the knee ligaments. In addition, the osseous morphology of the mouse knee was examined using high-resolution nanofocus computed tomography (CT), while standard micro-CT was employed to measure bone morphometrics of the distal femur and proximal tibia. Collectively, our findings suggest that the gross anatomy of the mouse knee is similar to the human knee despite some minor differences and features unique to the murine knee. The ACL had the highest load to failure (5.60 ± 0.75 N), the MCL (3.33 ± 1.45 N), and the PCL (3.45 ± 0.84 N) were similar, and the LCL (1.44 ± 0.37 N) had the lowest load to failure and stiffness. Murine models provide a unique opportunity to focus on biological processes that impact ligament pathology and healing due to the availability of transgenic strains. Our data support their use as a translational platform for the in vivo study of ligament injury, repair, and reconstruction., Competing Interests: None., (Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.)

A three-dimensional (3D) prosthesis was designed and built for a red-lored Amazon parrot ( Amazona autumnalis) with a pre-existing amputation of the distal left leg at the tibiotarsal-tarsometatarsal joint and injuries to the right leg caused by cage companion aggression. The prosthesis consisted of a straight main imprint, with a round element at both ends to provide stability, and a bridge connecting this to a socket without a bottom where the stump could be accommodated and held securely with self-adhesive bandaging. Over a 4-month period, 3 different 3D prosthetic models were made and evaluated. The first model was fitted, but the parrot would only use the tip of the main imprint to stand and walk. The second model was designed with a semicircular imprint with only 1 round element at the cranial end, a different bridge to accommodate the change to the main imprint, and the same socket. With these changes, the parrot was able to place the imprint of the prosthesis on the floor to stand and move freely around its enclosure. To accommodate morphologic changes on the stump, a third model was created consisting of the same imprint and bridge, but the socket was cut vertically all the way on one side to allow distention on its diameter and provide a long-lasting fit to the stump over time.

Purpose: To explore the microcirculation formation mechanism of contrast-enhanced (CE) ultrasonography imaging performance in rabbits with limb muscle crush injury., Methods: Seventy-two New Zealand white rabbits were randomly divided into two groups. A limb muscle crush injury model was created by airing a balloon cuff device with a force of 40 kpa. CE ultrasonography parameters were detected in the first group. In vivo microcirculation parameters were detected in the second group. Fine blood vessel diameter and blood flow velocity were calculated before extrusion and 0.5, 2, 6, 24 h, and 3 days after decompression., Results: Compared with the uninjured muscle, reperfusion of the injured muscles showed early and high enhancement in CE ultrasonography images. The time-intensity curve showed a trend of rapid elevation and gradual drop. Compared with the control group, fine artery and vein diameters in the experimental group were wider and the blood flow velocity was slower, especially in the fine veins., Conclusion: In vivo microcirculation detection can reflect changes in muscle microvascular diameter and blood flow velocity, which have a correlation with quantitative ultrasound imaging parameters.

Box-Behnken design (BBD) was performed via ultrasound-assisted system to optimize extraction yield and total antioxidant ability of polysaccharides from Suaeda fruticosa. Three independent variables were studied: temperature, time and pH with optimal settings 90 °C, 37 min and 2.9, respectively. Under these settings, isolation yield and total antioxidant activity were 34.0% and 31.8 mg ascorbic acid equivalent/g material, respectively. Furthermore, preliminary structure study was carried out using UV-vis analysis, Fourier transform infrared spectroscopy (FT-IR) and Nuclear magnetic resonance (1D ( 1 H, 13 C NMR) and 2D ( 1 H, 1 H COSY, 1 H/ 13 C HSQC)) methods Size exclusion chromatography (SEC) determination revealed that polysaccharide extracted had an average molecular weight of 240 kDa. Chemical composition analyses showed that extracted polysaccharide was a pectin-like polysaccharide with uronic acid content (47.5%) and neutral monosaccharides (52.5%) constituted mainly of arabinose, mannose, galactose, rhamnose, glucose and xylose in the molar percentage of 48.63%, 5.85%, 29.42%, 7.96%, 6.54% and 1.60%, respectively. This sugar composition was typical of that of a galactoarabinan. Importantly, the optimum polysaccharide possesses significant potentials in antioxidant activity effect. Moreover, pharmacological investigation of extracted polysaccharide revealed a good anti-inflammatory effect and interesting peripheral as well as central antinociceptive activities., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Three fucosylated chondroitin sulfate (fCS) were obtained from edible sea cucumbers Apostichopus japonicus, Stichopus chloronotus and Acaudina molpadioidea collected from China. fCS from Stichopus chloronotus was firstly reported. The detailed structures of fCSs, particularly the fucose branches, were investigated and compared. 1 H and 13 C NMR of the polysaccharide identified three sulfation patterns of fucose branches: 4-O-, 2,4-di-O, and 3,4-di-O-sulfation variously existed in different fCSs. The backbone structure was confirmed by the monosaccharide composition and two-dimensional NMR. Antioxidant properties of fCSs were evaluated by the scavenging abilities on 1,1-diphenyl-2-picrylhydrazyl, nitric oxide radicals and inhibition of lipid peroxidation. The results showed that their activities could be affected by the sulfation patterns of the fucose branches, and O-4 sulfation is particularly important for its activities. The anti-inflammatory assays of fCS-Am showed significant reduction of the carrageenan induced edema in a dose depended manner, which could be used as a potential antiallergic agent., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

OBJECTIVE To evaluate outcome of limb fracture repair in rabbits. DESIGN Retrospective case series. ANIMALS 139 client-owned rabbits with limb fractures treated between 2007 and 2015. PROCEDURES Medical records were reviewed for information on fracture location, fracture treatment, and time to fracture healing. RESULTS 25 rabbits had fractures involving the distal aspects of the limbs (ie, metacarpal or metatarsal bones, phalanges, and calcaneus or talus). Fractures were treated in 23 of these 25 rabbits (external coaptation, n = 17; external skeletal fixation, 4; and intramedullary pinning, 2) and healed in all 23, with a median healing time of 28 days (range, 20 to 45 days). One hundred ten rabbits had long bone fractures, and fractures were treated in 100 of the 110 (external skeletal fixation, n = 89; bone plating, 1; intramedullary pinning, 3; and external coaptation, 7). The percentage of fractures that healed was significantly lower for open (14/18) than for closed (26/26) tibial fractures and was significantly lower for femoral (19/26) and treated humeral (4/6) fractures than for radial (23/24) or closed tibial (26/26) fractures. Micro-CT was used to assess fracture realignment during external skeletal fixator application and to evaluate fracture healing. CONCLUSIONS AND CLINICAL RELEVANCE The prognosis for rabbits with limb fractures was good, with fractures healing in most rabbits following fracture repair (109/123). Micro-CT was useful in assessing fracture realignment and evaluating fracture healing.

OBJECTIVE To identify potential risk factors for digit injuries in dogs training and competing in agility events. DESIGN Internet-based, retrospective, cross-sectional survey. ANIMALS 1,081 dogs training or competing in agility events. PROCEDURES Data were collected for eligible animals via retrospective surveys distributed electronically to handlers of dogs participating in agility-related activities. Variables evaluated included demographic (handlers) and signalment (dogs) information, physical characteristics of dogs, and injury characteristics. A separate survey of dogs competing in similar agility-related activities but without digit injuries was also administered. Multivariable logistic regression was used to develop a model for assessment of risk factors. RESULTS Data were collected from 207 agility dogs with digit injuries and 874 agility dogs without digit injuries. Factors associated with significantly increased odds of injury included Border Collie breed (OR, 2.3; 95% confidence interval [CI], 1.5 to 3.3), long nails (OR, 2.4; 95% CI, 1.3 to 4.5), absence of front dewclaws (OR, 1.9; 95% CI, 1.3 to 2.6), and greater weight-to-height ratio (OR, 1.5; 95% CI, 1.1 to 2.0). Odds of injury decreased with increasing age of the dog (OR, 0.8; 95% CI, 0.76 to 0.86). CONCLUSIONS AND CLINICAL RELEVANCE Results should be cautiously interpreted because of potential respondent and recall bias and lack of review of medical records. Nevertheless, results suggested that retaining healthy dewclaws, maintaining lean body mass, and trimming nails short for training and competition may decrease the likelihood of digit injuries. Research to investigate training practices, obstacle construction specifcations, and surface considerations for dogs competing in agility activities is indicated.

Background: Adequate irrigation of open musculoskeletal injuries is considered the standard of care to decrease bacterial load and other contaminants. While the benefit of debris removal compared with the risk of further seeding by high-pressure lavage has been studied, the effects of irrigation on muscle have been infrequently reported. Our aim in the present study was to assess relative damage to muscle by pulsatile lavage compared with bulb-syringe irrigation., Methods: In an animal model of heterotopic ossification, 24 Sprague-Dawley rats underwent hindlimb blast amputation via detonation of a submerged explosive, with subsequent through-the-knee surgical amputation proximal to the zone of injury. All wounds were irrigated and underwent primary closure. In 12 of the animals, pulsatile lavage (20 psi [138 kPa]) was used as the irrigation method, and in the other 12 animals, bulb-syringe irrigation was performed. A third group of 6 rats did not undergo the blast procedure but instead underwent surgical incision into the left thigh muscle followed by pulsatile lavage. Serial radiographs of the animals were made to monitor the formation of soft-tissue radiopaque lesions until euthanasia at 6 months. Image-guided muscle biopsies were performed at 8 weeks and 6 months (at euthanasia) on representative animals from each group. Histological analysis was performed with hematoxylin and eosin, alizarin red, and von Kossa staining on interval biopsy and postmortem specimens., Results: All animals managed with pulsatile lavage, with or without blast injury, developed soft-tissue radiopaque lesions, whereas no animal that had bulb-syringe irrigation developed these lesions (p = 0.001). Five of the 12 animals that underwent blast amputation with pulsatile lavage experienced wound complications, whereas no animal in the other 2 groups experienced wound complications (p = 0.014). Radiopaque lesions appeared approximately 10 days postoperatively, increased in density until approximately 16 weeks, then demonstrated signs of variable regression. Histological analysis of interval biopsy and postmortem specimens demonstrated tissue damage with inflammatory cells, cell death, and dystrophic calcification., Conclusions: Pulsatile lavage of musculoskeletal wounds can cause irreversible insult to tissue, resulting in myonecrosis and dystrophic calcification., Clinical Relevance: The benefits and offsetting harm of pulsatile lavage (20 psi) should be considered before its routine use in the management of musculoskeletal wounds.

This study investigated the occurrence of wounds in Nigerian horses. The study population was 1,621 horses sold at the Obollo Afor horse lairage in Enugu State, Nigeria, during a 6-month period: 3 months of dry season and 3 months of rainy season (February-April and June-August 2012). A total of 207 horses were systematically sampled and subjected to a comprehensive physical examination. Those with wounds were marked, recorded, and clinically examined. Of the 207 horses sampled, 21 (10.1%) had wounds. The body distribution of the wounds was 9.5% head, 9.5% forelimbs, 19.1% hind limbs, 4.8% tail, 14.3% flank, 9.5% loin, 19.1% hip, 9.5% barrel, and 4.8% croup. The occurrence of the wounds was not significantly associated with sex or season, but the occurrence in adults was significantly (p < .05) higher than that in the young and aged horses. It was concluded that the occurrence of wounds is relatively high (10.1%), and mainly the hind limbs, hip, and flank of adult horses are affected. It was recommended that horse guardians and handlers should be properly educated on the care of horses.

Glycans are known to play important roles in molecular recognition, cell-cell adhesion, molecular trafficking, receptor activation, and signal transduction during development and regeneration. Despite numerous investigations of regenerating salamander limbs, global analysis of the precise variation of glycans during the limb regeneration process has received little attention. Here, we have used lectin microarrays and lectin histochemistry to analyze the alterations and distribution of glycans during the early stages leading to blastema formation during Cynops orientalis limb regeneration in response to limb amputation. Compared with the control group, analysis at several time points (3, 7, and 14 days postamputation) using microarrays containing 37 lectins showed that limb tissues expressed significantly different complements of glycans recognized by 9 different lectins. Postamputation limb tissues showed higher expression of two glycan structures recognized by the lectins STL and LTL and lower expression of seven glycan structures recognized by PHA-E, MAL-I, SNA, UEA-I, PHA-E + L, VVA, and GNA. We also observed significant changes in glycans specifically at 7 days postamputation, including higher binding capacity by WFA, GSL-I, and NPA and lower binding capacity by PNA, HHL, ConA, LCA, GSL-II, and PWM. Next, we validated our lectin microarray data using lectin histochemistry in limb tissues. Glycans recognized by STL and GNA showed similar changes in signal intensity to those found in the lectin microarrays, with STL staining in the cytoplasm and GNA in the cytoplasm and nucleus. Our findings are the first report of significant postamputation changes in glycans in limb tissues and suggest that those glycans perform potentially important functions during the early stages of C. orientalis limb regeneration., (© 2017 by the Wound Healing Society.)

Mouse digit tip regeneration involves an intricate coordinated regrowth of the terminal phalanx, nail, dermis and epidermis. During this time, regenerating digits undergo wound healing, blastema formation, and differentiation. However, the regenerative response of the digit is dependent on the level of the amputation. Amputation of <30% of the distal phalanx (P3), with part of the base nail remaining, results in extensive digit regeneration. In contrast, >60% P3 removal results in no regeneration. This level-dependent regenerative ability of the mouse digit provides a comparative model between regeneration and non-regeneration that may enable identification of specific factors critical to regeneration. Although the ability to create regenerating and non-regenerating conditions has been well established, the regenerative response between these regions ("intermediate" zone) has received less scrutiny, and may add insight to the regenerative processes, including the degree of histolysis, and the level of blastema formation. The objective of this study is then to compare the regeneration capacity between amputation levels within the regenerating (<30%), intermediate (40-59%), and non-regenerating (>60%) regions. Results indicated that regenerative and intermediate amputations led to significant histolysis and blastema formation of the distal phalanx 14 days post-amputation. Unlike the regenerating digits, intermediate amputations led to incomplete regeneration whereby regrowth of the digits were not to the levels of the intact or regenerating digits. Non-regenerating amputations did not exhibit significant histolysis or blastema formation. Remarkably, the histolytic process resulted in day 14 P3 lengths that were similar regardless of the initial amputation over 19%. The differences in histolysis, blastema formation and injury outcomes were also marked by changes in the number of proliferating cells and osteoclasts. Altogether, these results indicate that although intermediate amputations result in histolysis and blastema formation similar to regenerating digits, the resulting cellular composition of the blastema differs, contributing to incomplete regeneration., (© 2017 by the Wound Healing Society.)

According to the Agreement on International Humane Trapping Standards (AIHTS), which was signed by the European Community, Canada, and Russia in 1997, killing devices used for the capture of canids and other fur-bearing nonhuman animals should render an animal irreversibly unconscious within 300 s. However, killing neck snares are not included in the agreement. In this commentary, a parallel is drawn between injuries caused by steel-jawed leghold traps, which have been banned by the AIHTS signatory countries, and killing neck snares to demonstrate that these snares should also be included in international humane trapping standards (i.e., AIHTS). Previous scientific investigations have shown that neither manual nor power-killing neck snares can consistently render canids unconscious rapidly. Animals caught in killing neck snares suffer injuries that are similar to or worse than those reported for leg-captured canids. The authors strongly recommend that AIHTS be modified to include killing neck snares and that such devices be subject to the criteria applied to other trapping devices. Alternative restraining trapping devices, which are effective and more humane, are available for capturing wild canids.

Background: Traumatic bone fractures cause moderate to severe pain, which needs to be minimized for optimal recovery and animal welfare, illustrating the need for reliable objective pain biomarkers for use in a clinical setting. The objectives of this study were to investigate catestatin (CST) and vasostatin (VS) concentrations as two new potential biomarkers, and cortisol concentrations, scores of the short form of the Glasgow composite measure pain scale (CMPS-SF), and visual analog scale (VAS) in dogs suffering from traumatic bone fractures before and after morphine administration in comparison with healthy dogs., Methods: Fourteen dogs with hind limb or pelvic fractures and thirty healthy dogs were included. Dogs with fractures were divided into four groups according to analgesia received before participation. Physical examination, CMPS-SF, pain and stress behavior VAS scores were recorded in all dogs. Saliva and blood were collected once in healthy dogs and in dogs with fractures before and 35-70 min after morphine administration. Blood samples were analyzed for CST, VS, and cortisol. Saliva volumes, however, were insufficient for analysis., Results: Catestatin and cortisol concentrations, and CMPS-SF, and VAS scores differed significantly between dogs with fractures prior to morphine administration and healthy dogs. After morphine administration, dogs with fractures had significantly decreased CMPS-SF and VAS scores and, compared to healthy dogs, CST concentrations, CMPS-SF, and VAS scores still differed significantly. However, CST concentrations remained largely within the normal range. Absolute delta values for CST significantly correlated with delta values for CMPS-SF. Catestatin and cortisol did not differ significantly before and after morphine administration. Vasostatin concentrations did not differ significantly between groups., Conclusions: Catestatin and cortisol concentrations, CMPS-SF, and VAS scores differed significantly in the dogs with traumatic bone fractures compared to the healthy dogs. Morphine treatment partially relieved pain and stress according to the subjective but not according to the objective assessments performed. However, because of the large degree of overlap with normal values, our results suggest that plasma CST concentrations have a limited potential as a clinically useful biomarker for pain-induced stress.

The NLRP3 inflammasome is a multi-protein complex that assembles in response to tissue damage or infection, triggering activation of caspase-1, an enzyme that converts interleukin (IL)-1β into its active form. A role for the NLRP3 inflammasome is emerging in inflammatory pain, but its influence in other pain types is largely unexamined. Therefore the aim of this study was to assess the role of the NLRP3 inflammasome and its downstream product caspase-1 in a model of acute burn-induced pain in male mice. A superficial burn was induced on the plantar surface of the left hind paw using a hot plate set at 52.5°C for 25s. Development of burn-induced mechanical allodynia, thermal allodynia, edema and weight bearing changes was assessed in Nlrp3 -/- and caspase-1-deficient (Ice -/- ) mice, and in mice administered the selective NLRP3 inflammasome inhibitor MCC950. Burn-induced mechanical and thermal allodynia developed normally in Nlrp3 -/- and Ice -/- mice and mice administered MCC950. Burn-induced edema was significantly reduced in Ice -/- mice only. Burn-induced weight bearing changes were attenuated in Nlrp3 -/- mice and mice administered MCC950 72h after burn only. This study suggests that NLRP3 and its downstream product caspase-1 have a limited role in the development of burn-induced pain., (Copyright © 2016 Elsevier Ltd and ISBI. All rights reserved.)

Objective: To describe the diagnosis, treatment, and prognosis of fetlock breakdown due to interosseus medius muscle rupture in cattle., Study Design: Retrospective clinical study., Animals: Dairy heifers with unilateral or bilateral interosseus medius muscle rupture (n = 11)., Methods: Breakdown injury due to rupture of the interosseus medius muscle was documented clinically, radiographically, and ultrasonographically. Breakdown was bilateral in 4 heifers (3 in forelimbs, 1 in hind limbs) and unilateral in 7 (all hind limbs). One heifer with severe bilateral hind limb breakdown was slaughtered and the remaining 10 were treated by transfixation pin cast (1 heifer), box rest (n = 2), and/or a cast and splint (7)., Results: Hyperextension of the fetlock and hyperflexion of the proximal interphalangeal joints during weight bearing were characteristic for interosseus muscle breakdown. Ultrasonographically, the origin and body of the interosseus muscle and the branches to the sesamoid bones were primarily affected by the rupture. Conservative treatment was successful (used for their intended purpose) in 8 of 9 heifers with a median lifespan of 32 months after discharge from the clinic (range 6-83)., Conclusion: Rupture of the interosseus medius muscle in young cattle may be more common than previously suggested in the literature. Imaging with ultrasound allowed more detailed localization of lesions of the musculo-tendinous structure. Interosseus medius muscle rupture had a favorable prognosis when treated conservatively in these heifers., (© 2017 The American College of Veterinary Surgeons.)

Cortical perception of noxious stimulation is an essential component of pain experience but it is not known how cortical nociceptive activity emerges during brain development. Here we use continuous telemetric electrocorticogram (ECoG) recording from the primary somatosensory cortex (S1) of awake active rat pups to map functional nociceptive processing in the developing brain over the first 4 weeks of life. Cross-sectional and longitudinal recordings show that baseline S1 ECoG energy increases steadily with age, with a distinctive beta component replaced by a distinctive theta component in week 3. Event-related potentials were evoked by brief noxious hindpaw skin stimulation at all ages tested, confirming the presence of functional nociceptive spinothalamic inputs in S1. However, hindpaw incision, which increases pain sensitivity at all ages, did not increase S1 ECoG energy until week 3. A significant increase in gamma (20-50 Hz) energy occurred in the presence of skin incision at week 3 accompanied by a longer-lasting increase in theta (4-8 Hz) energy at week 4. Continuous ECoG recording demonstrates specific postnatal functional stages in the maturation of S1 cortical nociception. Somatosensory cortical coding of an ongoing pain "state" in awake rat pups becomes apparent between 2 and 4 weeks of age., (© The Author 2016. Published by Oxford University Press.)

Glucosamine (GlcN) fulfills many of the requirements as an ideal component in scaffolds used in cartilage tissue engineering. The incorporation of GlcN in a gelatin/hyaluronic acid (GH) cryogel scaffold could provide biological cues in maintaining the phenotype of chondrocytes. Nonetheless, substituting gelatin with GlcN may also decrease the crosslinking density and modulate the mechanical properties of the cryogel scaffold, which may be beneficial as physical cues for chondrocytes in the scaffold. Thus, we prepared cryogel scaffolds containing 9% GlcN (GH-GlcN9) and 16% GlcN (GH-GlcN16) by carbodiimide-mediated crosslinking reactions at -16 °C. The crosslinking density and the mechanical properties of the cryogel matrix could be tuned by adjusting the content of GlcN used during cryogel preparation. In general, incorporation of GlcN did not influence scaffold pore size and ultimate compressive strain but increased porosity. The GH-GlcN16 cryogel showed the highest swelling ratio and degradation rate in hyaluronidase and collagenase solutions. On the contrary, the Young's modulus, storage modulus, ultimate compressive stress, energy dissipation level, and rate of stress relaxation decreased by increasing the GlcN content in the cryogel. The release of GlcN from the scaffolds in the culture medium of chondrocytes could be sustained for 21 days for GH-GlcN16 in contrast to only 7 days for GH-GlcN9. In vitro cell culture experiments using rabbit articular chondrocytes revealed that GlcN incorporation affected cell proliferation, morphology, and maintenance of chondrogenic phenotype. Overall, GH-GlcN16 showed the best performance in maintaining chondrogenic phenotype with reduced cell proliferation rate but enhanced glycosaminoglycans (GAGs) and type II collagen (COL II) secretion. Quantitative real-time polymerase chain reaction also showed time-dependent up-regulation of cartilage-specific marker genes (COL II, aggrecan and Sox9) for GH-GlcN16. Implantation of chondrocytes/GH-GlcN16 constructs into full-thickness articular cartilage defects of rabbits could regenerate neocartilage with positive staining for GAGs and COL II. The GH-GlcN16 cryogel will be suitable as a scaffold for the treatment of articular cartilage defects., Competing Interests: The authors declare no conflict of interest.

Objectives: The aim of this study was to assess retrospectively the efficacy and complication rate of hindlimb peripheral nerve blocks (PNBs) in cats., Methods: Clinical records of cats that received PNBs and underwent hindlimb orthopaedic surgery from February 2010 to October 2014 were examined. Type of PNB, type and dose of local anaesthetic used, end-expiratory fraction of isoflurane (FE'Iso) administered, additional intraoperative analgesia, incidence of hypotension, postoperative opioid requirement, postoperative contralateral limb paralysis and neurological complications at the 6 week re-examination were investigated., Results: Eighty-nine records were retrieved but only 69 were analysed. Four combinations of PNBs were used: 34 lateral preiliac (LPI) approach to lumbar plexus (LP) associated with lumbar paravertebral approach to sciatic nerve (SN); 20 LPI-LP associated with the lateral approach to SN; three LPI-LP associated with gluteal approach to SN; 12 dorsal-paravertebral (DPV) approach to LP associated with lateral SN. Levobupivacaine was used for the majority of PNBs. The mean intraoperative FE'Iso was 1.15%; hypotension was documented in 55.1% of anaesthetics, while 31.8% of cats received fentanyl and/or ketamine intraoperatively. Postoperatively, 72.7% of cats received at least one dose of opioid, while five cats required further postoperative analgesia (ketamine constant rate infusion and/or gabapentin). No cats showed contralateral limb paralysis and neurological complications at the 6 week re-examination. No differences were found when comparing the different PNBs used., Conclusions and Relevance: PNBs contributed to perioperative anaesthesia/analgesia in cats undergoing hindlimb orthopaedic surgery. However, the clinical relevance of intraoperative hypotension needs further investigation., (© The Author(s) 2015.)

Ethiopia has 7.1 million donkeys and mules, the majority of which are used as pack animals. Factors such as poor harness quality, long-distance traveling, and heavy cartloads have been linked to reduced work efficiency. Addressing the health and welfare of working equids is imperative not only for the animals but also for the households dependent upon them for livelihood. In developing countries, 75 % of working equids have gait or limb abnormalities, but the relationship between workload and prevalence of lameness is unknown. We examined 450 cart mules in Bahir Dar, Ethiopia. Lameness and workload were assessed through use of a survey and lameness exam. We found that 26.8 % of cart mules were lame, and acute lameness of the forelimb was the most common. Animals with poor harness quality were 2.5 times more likely to have sores and 1.6 times more likely to be lame. Lameness tended to be associated with cartloads >700 kg (P = 0.09), and there was a significant association between multiple-leg lameness and cartload weight (P = 0.03). The presence of sores was the best predictor of lameness (P = 0.001). Possible areas of intervention may include education to reduce average daily workload and improving harness design.