The use of a laparoscopic suction-irrigation device in 2 standing horses for lavage of the abdomen for the treatment of primary suppurative peritonitis is reported. Two horses were presented with a 1- to 2-week history of weight loss. Abdominocentesis revealed highly elevated total nucleated cell count. Peritoneal lavage systems were placed in both horses, but complications prevented adequate lavage. Both horses underwent standing laparoscopy; the dorsal abdomen was explored and the abdomen was profusely lavaged, using a suction-irrigation device. The procedure was efficient and allowed adequate visualization of the dorsal abdomen and lavage. A successful outcome was achieved in both cases. Key clinical message: Lavage of the abdomen of horses with peritonitis can be achieved under standing sedation, using a laparoscopic technique. In appropriately selected cases, this allows for adequate visualization of the dorsal abdomen and efficacious abdominal lavage.

Critical sized bone defect (CSBD) animal models are used to evaluate and confirm efficacy and potency of new treatment modalities based on bone tissue engineering before the latter can be applied in clinical practice. In this study, a bilateral CSBD model in the iliac wings of sheep is described in detail. To demonstrate that this is a large animal CSBD model in sheep, bone healing within the defect left empty (negative control) or filled with autologous corticocancellous bone graft (clinical gold standard, positive control) was assessed using micro-CT, histology, histomorphometric, and fluorochrome analysis. After three months, new bone into the defect site was formed across the whole defect in the positive controls but limited to the edge of the defects in the negative controls. Bone volume in the positive controls was statistically higher than in the negative controls, with the latter having less than 10% new bone growth. There were no intraoperative or postoperative complications. The model described here represents a reliable and reproducible bilateral CSBD in sheep with low morbidity that can be used for in vivo evaluation of new treatment modalities based on bone tissue engineering.

Bone substitutes, like calcium phosphate, are implemented more frequently in orthopaedic surgery to reconstruct critical size defects, since autograft often results in donor site morbidity and allograft can transmit diseases. A novel bone cement, based on β-tricalcium phosphate, polyethylene glycol, and trisodium citrate, was developed to allow the rapid manufacturing of scaffolds, by extrusion freeform fabrication, at room temperature. The cement composition exhibits good resorption properties and serves as a basis for customised (e.g., drug or growth factor loaded) scaffolds for critical size bone defects. In vitro toxicity tests confirmed proliferation and differentiation of ATDC5 cells in scaffold-conditioned culture medium. Implantation of scaffolds in the iliac wing of sheep showed bone remodelling throughout the defects, outperforming the empty defects on both mineral volume and density present in the defect after 12 weeks. Both scaffolds outperformed the autograft filled defects on mineral density, while the mineral volume present in the scaffold treated defects was at least equal to the mineral volume present in the autograft treated defects. We conclude that the formulated bone cement composition is suitable for scaffold production at room temperature and that the established scaffold material can serve as a basis for future bone substitutes to enhance de novo bone formation in critical size defects.

OBJECTIVE:To compare the torsional strength of calf metatarsal bones with defects produced by removal of 2 different implants.STUDY DESIGN:In vitro mechanical comparison of paired bones with bicortical defects resulting from the implantation of 2 different external fixation systems: the transfixation pin (TP) and the pin sleeve system (PS).SAMPLE POPULATION:Neonatal calf metatarsal bones (n = 6 pairs).METHODS:From each pair, 1 bone was surgically instrumented with 2 PS implants and the contralateral bone with 2 TP implants. Implants were removed immediately leaving bicortical defects at identical locations between paired metatarsi. Each bone was tested in torque until failure. The mechanical variables statistically compared were the torsional stiffness, the torque and angle at failure, and work to failure.RESULTS:For TP and PS constructs, respectively, there were no significant differences between construct types for any of the variables tested. Mean ± SD torsional stiffness: 5.50 ± 2.68 and 5.35 ± 1.79 (Nm/°), P = .75; torque: 57.42 ± 14.84 and 53.43 ± 10.16 (Nm); P = .34; angle at failure: 14.76 ± 4.33 and 15.45 ± 4.84 (°), P = .69; and work to failure 7.45 ± 3.19 and 8.89 ± 3.79 (J), P = .17).CONCLUSIONS:Bicortical defects resulting from the removal of PS and TP implants equally affect the investigated mechanical properties of neonate calf metatarsal bones.

Bone grafts are commonly used for the treatment of segmental bone defects and fracture non-unions. Recently, osseous particles obtained during intermedullary canal reaming (using a Reamer-Irrigator-Aspirator (RIA) device) have been evaluated as graft material during in vitro and clinical studies. The aim of this study was to evaluate and quantify new bone formation after implantation of bone graft material obtained after reaming of the tibia in a bilateral critical-sized iliac wing defect in sheep and to investigate the effect of the augmentation of this graft. A reamer bone graft alone, or after short term incubation in a dexamethasone enriched solution, and a reamer graft collected using beta-tricalcium phosphate (β-TCP) granules in the filter of the RIA collection device were compared to autologous iliac wing graft. In addition, reamer graft was combined with the cellular fraction collected from the irrigation fluid with and without short-term incubation in a dexamethasone enriched solution. It was hypothesized that the amount of physical bone in the reamer bone graft groups would be higher than the amount in the autologous iliac wing graft group and that augmentation of a reamer bone graft would increase bone formation. Three months after implantation, the amount of new bone formation (as percentage of the total defect volume) in the defects was evaluated ex-vivo by means of micro-CT and histomorphometry. The mean amount of bone in the autologous iliac wing graft group was 17.7% and 16.8% for micro-CT and histomorphometry, respectively. The mean amount of bone in all reamer graft groups ranged between 20.4–29.2% (micro-CT) and 17.0–25.4% (histomorphometry). Reamer graft collected using β-TCP granules (29.2 ± 1.7%) in the filter produced a significantly higher amount of bone in comparison to an autologous iliac wing graft evaluated by micro-CT. RIA bone grafts added a small increase in bone volume to the 3 month graft volume in this preclinical sheep model. The current model does not support the use of short-term high concentration dexamethasone for augmentation of a graft volume. If avoidance of an iliac wing graft is desirable, or a reaming procedure is required, then a RIA graft or RIA graft plus β-TCP granules are as good as the current gold standard for this model.

Minimally invasive surgery, including laparoscopy, thoracoscopy, and endoscopic-assisted procedures, is becoming more common in veterinary medicine. The basic principles of laparoscopy and thoracoscopy involve gaining access to a cavity through small incisions, creating a workspace, and maneuvering extracorporeally activated instruments within that workspace.

OBJECTIVE To compare proximal fragment displacement and the peri-implant strain using a pin-sleeve cast (PSC) system and a transfixation pin cast (TPC) system on a cadaveric calf metacarpal bone fracture model. STUDY DESIGN Experimental. SAMPLE POPULATION Cadaveric calf metacarpal bones (n = 6 pairs). METHODS Paired samples were instrumented with either the TPC or the PSC systems. Strain gauges were applied proximal to the transfixation implants and the bones encased in cast material. The distal part of the construct was removed to mimic an unstable distal comminuted fracture. Constructs were fixed to the material testing machine and initially loaded in axial compression in their elastic range to determine construct stiffness. Constructs were loaded cyclically with a sinusoidal curve that increased until failure. Variables compared statistically between constructs were the initial construct stiffness and, at given load points, the mean metacarpal axial displacement in loading and unloading condition and mean axial strain. RESULTS Initial construct mean ± SD axial stiffness was not significantly different between constructs (PSC: 689 ± 258; TPC: 879 ± 306 N/mm). There was no significant difference between either investigated displacements of metacarpal bones transfixed with PSC and those transfixed with TPC at all load points. The PSC constructs had a significant decrease in the recorded mean strain (502 ± 340 μstrain) compared to the TPC construct (1738 ± 2218 μstrain). CONCLUSIONS The PSC significantly reduced peri-implant strain with comparable axial displacement to the TPC in cadaveric calf metacarpal bones.

The osteointegration of a new three-dimensional reticular titanium material, Trabecular Titanium™, was assessed using a bilateral cancellous (distal femur, proximal tibia) and cortical (tibia diaphysis) bone drill hole model in 18 sheep. TT is a novel Ti6Al4V material characterized by a high open porosity and composed of multi-planar regular hexagonal cells. Two 5.0 mm diameter, 12 mm long cylinders (TT1 & TT2) of two different porosities (TT1:650 μm, TT2:1250 μm) were tested and compared to two solid predicate 5.0 mm diameter, 12 mm long Ti cylinders (PT1 & PT2) coated with porous Ti (PT1: vacuum-plasma spray coating; PT2: inert-gas shielding arc spray coating). Each implant type was surgically implanted at 4 separate locations in each sheep (16 implants per sheep). Three timepoints of 4, 16 and 52 weeks (n=6 sheep per timepoint) were used. Bone-implant interface was analyzed ex vivo by the determination of: 1) the shear strength (SS) measured during a push out test, 2) the percentage of bone in-growth (%B) using histomorphometry, 3) the bone apposition rate using fluorochrome labelling analysis and 4) the bone-implant contact using backscattered scanning electron microscopy (SEM). An ANOVA with a Bonferroni Post hoc test were used to detect differences between tested and predicate implants. P values 0.05 were considered significant. At 4 weeks, 5 out of the 6 TT1 could be pushed out of the cortical bone (COB) samples. The remaining TT1 collapsed during testing. All TT1 could be pushed of the cancellous bone (CAB) samples. Four out of the 6 TT2 could be pushed out of CAB and of the COB samples. At 16 and 52 weeks, only one TT1 and one TT2 could be pushed out of the bone samples, the remaining implants collapsed during testing. All the PTs were successfully pushed out at all timepoints. The mean %B of PT1 and PT2 did not significantly increase over time. For both materials, the mean %B ranged between 1.7% and 4.4% at 4 weeks and between 5.7% and 6.5% at 52 weeks. The mean %B of TT1 significantly increased over time in both COB (10.2% at 4 weeks, 46.2% at 16 weeks, 50.5% at 52 weeks) and CAB (5.8%, 23.9%, 24.3%). Similarly, the mean %B of TT2 significantly increased over time in both COB (7.8%, 48.6%, 65%) and CAB (4.5%, 24.1%, 38.6%). Bone apposition rates for the TT implants remained superior to 2 μm/day for the entire duration of the study. SEM showed an intimate bone-implant contact for all implant types at all timepoints. At 16 and 52 weeks, histomorphometry revealed an extensive osteointegration of the TT specimens. Bone-implant interface strength was so high for the TT implants that they could not be pushed out of the bone samples. The results of this study would indicate that the TT implants provide a good scaffold for bone in-growth.

Laparoscopy, thoracoscopy, and other endoscopic-assisted procedures have many uses in veterinary medicine. Diagnosis and biopsy are currently the most common uses of minimally invasive techniques, but popularity for procedures such as ovariectomy, cancer staging, lung lobectomy, and pericardectomy is rising. This article touches on basic surgical procedures and describes some more advanced techniques.

OBJECTIVETo evaluate 2 plate designs for pancarpal arthrodesis and their effects on load transfer to the respective bones as well as to develop a computational model with directed input from the biomechanical testing of the 2 constructs.SAMPLEBoth forelimbs from the cadaver of an adult castrated male Golden Retriever.PROCEDURESCT imaging was performed on the forelimb pair. Each forelimb was subsequently instrumented with a hybrid dynamic compression plate or a castless pancarpal arthrodesis plate. Biomechanical testing was performed. The forelimbs were statically loaded in the elastic range and then cyclically loaded to failure. Finite element (FE) modeling was used to compare the 2 plate designs with respect to bone and implant stress distribution and magnitude when loaded.RESULTSCyclic loading to failure elicited failure patterns similar to those observed clinically. The mean ± SD error between computational and experimental strain was < 15% ± 13% at the maximum loads applied during static elastic loading. The highest bone stresses were at the distal extent of the metacarpal bones at the level of the screw holes with both plates; however, the compression plate resulted in slightly greater stresses than did the arthrodesis plate. Both models also revealed an increase in bone stress at the proximal screw position in the radius. The highest plate stress was identified at the level of the radiocarpal bone, and an increased screw stress (junction of screw head with shaft) was identified at both the most proximal and distal ends of the plates.CONCLUSIONS AND CLINICAL RELEVANCEThe FE model successfully approximated the biomechanical characteristics of an ex vivo pancarpal plate construct for comparison of the effects of application of different plate designs.

Carriers for bone morphogenetic protein-2 (BMP-2) used in clinical practice still suffer from limitations such as insufficient protein retention. In addition, there is a clinical need for injectable carriers. The main objective of this study was to assess bone forming ability of rhBMP-2 combined either with chitosan hydrogel (rhBMP-2/CH) or chitosan hydrogel containing β-tricalcium phosphate (β-TCP) (rhBMP-2/CH/TCP). Formulations were first compared in a rat ectopic intramuscular bone formation model, and the optimal formulation was further evaluated in healing of 15-mm critical size defect in the radius of a rabbit. Three weeks after injection ectopically formed bone was analyzed by microcomputerized tomography (micro-CT) and histology. Significantly higher (4.7-fold) mineralized bone formation was observed in the rhBMP-2/CH/TCP group compared to rhBMP-2/CH group. In a pilot study, defect in a rabbit radius treated with rhBMP-2/CH/TCP showed incomplete regeneration at 8 weeks with composite leakage from the defect, indicating the need for formulation refinement when segmental defect repair is foreseen.

The transfixation pin cast (TPC) is an external skeletal fixation technique used to treat horses with distal limb fractures, but its use is often associated with pin-loosening and an increased risk of treatment failure. To address implant loosening, the pin sleeve cast system (PSC) was recently designed and consists of a pin-sleeve unit inserted into the bone. Each pin runs through a sleeve placed in the bone, making contact at two fixed points only within the sleeve. Each pin is attached to a ring embedded in a resin cast. In this report, the mechanical performance of a traditional TPC pin arrangement was compared with that of the PSC using validated finite element models of bone substitutes previously tested in vitro. The PSC resulted in a marked reduction in peak strain magnitude around the pins and a more even distribution of strain across the bone cortex. The two systems resulted in comparable proximal fragment displacement and had a similar stress concentration around bone defects during implant removal. The findings suggest that the PSC load transfer mechanism is effective even in geometrically complex structures like equine bones.

Background and Aim Fractures of the tibia and femoral diaphysis are commonly repaired by intramedullary (IM) nails, which are currently composed of either electropolished stainless steel (EPSS) or standard, non-polished titanium-aluminum-niobium (TAN). Once the fracture has fully healed, removal of IM nails is common, but the strong adhesion of bone to standard TAN complicates removal. Polishing the surface of TAN IM nails has been shown to reduce bony adhesion and ease implant removal without compromising fixation. Polished TAN nails are, therefore, expected to have significant clinical benefit in situations where the device is to be removed. The aim of the present study was to determine the effect of polishing TAN IM nails on susceptibility to infection in an animal model. Materials and Methods Solid IM nails (Synthes, Betlach, Switzerland) composed of standard TAN were compared with polished equivalents and also to clinically available EPSS nails. The surface chemical and topographical properties of the materials were assessed by X-ray photon spectroscopy (XPS), white light profilometry, and scanning electron microscopy (SEM). An in vivo infection study was performed using a clinical isolate of Staphylococcus aureus that was characterized with respect to various virulence factors. Results Polishing TAN IM nails caused no significant change to the chemistry of the nails, but the topography of the polished TAN nails was significantly smoother than standard TAN nails. In the infection study, the rank order based on descending infectious dose 50 (ID50) was: standard TAN, polished TAN, and finally EPSS. The ID50 values did not differ greatly between any of the groups. Conclusions Polishing the surface TAN IM nails was not found to influence the susceptibility to infection in our animal model.

OBJECTIVE:To describe laparoscopic removal of a large testicular teratoma in a standing horse.STUDY DESIGN:Clinical report.ANIMALS:Thoroughbred horse (11 months) with a testicular teratoma.METHODS:A unilateral cryptorchid testicle could not be removed by an inguinal approach under general anesthesia because of it s large size. After recovery from general anesthesia, ultrasound evaluation revealed a 24 x 19 cm fluid-filled testicular mass. The mass was removed by paralumbar fossa laparoscopy with the horse in a standing position. After fluid aspiration of the mass, the mesorchium and ductus deferens were ligated with extracorporeal knots and the mass retrieved inside a laparoscopic specimen pouch. Morphologic features were consistent with a teratoma.RESULTS:Laparoscopic-guided aspiration of fluid from the teratoma decreased mass size and increased ease of manipulation and retrieval. Retrieval of the teratoma in a laparoscopic specimen pouch prevented loss of abdominal insufflation, helped reduce fluid leakage, and potential seeding of neoplastic cells.CONCLUSION:Use of laparoscopy for removal of neoplastic cryptorchid testicles offers many advantages including minimal invasiveness and increased safety associated with good visibility of structures.CLINICAL RELEVANCE:Standing laparoscopic surgery should be considered for removal of testicular neoplasms in horses.

OBJECTIVE:To evaluate the efficacy of a novel pin-sleeve cast (PSC) system for external fixation of distal limb fractures in horses and to compare it with the transfixation pin cast (TPC) system.STUDY DESIGN:Experimental.SAMPLE POPULATION:One bone substitute each was used for the TPC and PSC systems. The PSC was tested in 4 configurations characterized by different pin preloads.METHODS:Specimens were loaded in axial compression in the elastic range. Variables compared statistically were: bone substitute axial displacement and axial strain measured above implants with strain gauges. Pin preload was correlated with the variables investigated. Load to failure and a fatigue tests supplemented the investigation.RESULTS:The PSC configuration with the highest pin preload showed a significantly lower axial displacement compared with the TPC. No significant differences were observed between all other PSC configurations and the TPC. All PSC systems had a significant decrease in recorded strain compared with the TPC system. Pin axial preload inversely correlated with axial displacement but had no effect on axial strain. In the failure test, the PSC encountered plastic deformation earlier than the TPC. In the fatigue test, the PSC ran >200,000 cycles.CONCLUSIONS:Preliminary in vitro tests showed that the PSC system significantly reduced peri-implant strain while concurrently having comparable axial displacement to the TPC system.CLINICAL RELEVANCE:The PSC system has the potential to reduce the risk of pin loosening in horses.

Experimental and epidemiological evidence suggests that consumption of hydrolyzable carbohydrate, hCHO (grain), by horses is an important risk factor for colic, a common cause of equine mortality. It is unknown whether the small intestinal capacity to digest hCHO and/or to absorb monosaccharides is limiting, or even if horses can adapt to increased carbohydrate load. We investigated changes in the brush-border membrane carbohydrate digestive enzymes and glucose absorptive capacity of horse small intestine in response to increased hCHO. Expression of the Na(+)/glucose co-transporter, SGLT1, was assessed by Western blotting, immunohistochemistry, Northern blotting, QPCR, and Na(+)-dependent D-glucose transport. Glucose transport rates, SGLT1 protein, and mRNA expression were all 2-fold higher in the jejunum and 3- to 5-fold higher in the ileum of horses maintained on a hCHO-enriched diet compared to pasture forage. Activity of the disaccharidases was unaltered by diet. In a well-controlled study, we determined SGLT1 expression in the duodenal and ileal biopsies of horses switched, gradually over a 2-month period, from low (<1.0 g/kg bwt/day) to high hCHO (6.0 g/kg bwt/day) diets of known composition. We show that SGLT1 expression is enhanced, with time, 2-fold in the duodenum and 3.3-fold in the ileum. The study has important implications for dietary management of the horse.

OBJECTIVE:To assess the antimicrobial elution characteristics, toxicity, and antimicrobial activity of amikacin-impregnated ferric-hyaluronate implants (AI-FeHAI) for amikacin delivery to the tarsocrural joint of horses.STUDY DESIGN:Experimental study.SAMPLE POPULATION:AI-FeHAI implants, equine cartilage, and synovium, and horses (n=6).METHODS:In vitro study: Five AI-FeHAI were placed in saline solution with daily replacement until implant degradation. Eluent was tested for amikacin concentration and bioactivity. Synovial and cartilage explants were incubated in the presence or absence of AI-FeHAI for 72 hours and subsequently assessed for morphology, viability, and composition. Synovial explants were incubated with Staphylococcus aureus in the presence or absence of AI-FeHAI. Spent medium was cultured daily and explants were assessed for morphology and viability after 96 hours. In vivo study: AI-FeHAI were placed in 6 tarsocrural joints. Standard cytologic analysis and amikacin concentration (SFAC) were determined in synovia obtained regularly for 28 days thereafter. Similar analyses were conducted after a single intra-articular injection of amikacin 6 months later.RESULTS:In vitro study: Amikacin concentrations exceeded 16 microg/mL and inhibited S. aureus growth for 8 days. AI-FeHAI had no effect on cartilage explants. AI-FeHAI eliminated bacteria from synovial explants. In vitro study: After AI-FeHAI placement, SFAC was highest (140.78+63.81 microg/mL) at first sampling time. By 24 hours SFAC was <16 microg/mL. After intra-articular injection, SFAC was the highest (377.91 +/- 40.15 microg/mL) at first sampling time. By 48 hours SFAC was <16 microg/mL.CONCLUSIONS:A single intra-articular amikacin injection demonstrated superior pharmacokinetics than AI-FeHAI prepared as described.CLINICAL RELEVANCE:AI-FeHAI cannot be recommended for clinical use.