Ex vivo comparison of one-layer versus two-layer closures in end-to-end anastomoses in normal equine descending colon.

OBJECTIVE: The aim of the study was to evaluate time to closure, bursting pressures and luminal diameters of a single and double-layer end-to-end anastomoses in normal equine descending colon. STUDY DESIGN: Experimental study. SAMPLE POPULATION: Eight adult horses. METHODS: Four segments of descending colon from each horse were randomly assigned to a control (n = 8, CON), one-layer (n = 12, group 1; continuous Lembert pattern), or two-layer group (n = 12, group 2; simple continuous oversewn with Cushing pattern). Anastomoses were performed, and time to closure and luminal diameter were measured. Bursting pressures were determined, and location of failure was recorded. Mixed analysis of variance (ANOVA) was employed. RESULTS: Mean time to completion was decreased (p = .003) in group 1 (18.6 min ± 22.8 s) compared with group 2 (21.35 min ± 22.8 s). Luminal diameter was reduced (p < .0001) in group 2 (47 ± 2.46 mm) compared to group 1 (65 ± 2.77 mm). Location of failure was remote to the anastomosis in all but one segment; therefore, differences in anastomotic bursting pressures could not be determined. CONCLUSION: Two-layer closures resulted in significant luminal reduction and took longer to complete than one-layer anastomoses. CLINICAL SIGNIFICANCE: Use of one-layer closures may be advantageous primarily with respect to luminal diameter.

Effect of repeated steam sterilizations on insertional torque, torque to failure, and axial pullout strength of 3.5-mm and 2.0-mm cortical bone screws.

OBJECTIVE: The objective of this study was to determine the effects of repeated steam sterilization cycles on the biomechanical properties of surgical screws. METHODS: 42 3.5-mm and 42 2.0-mm self-tapping, cortical screws were divided into 3 groups per size and underwent autoclave sterilization for 1 (G1), 50 (G50), or 100 (G100) cycles and testing from August 2018 through June 2021. Sixty screws were then inserted into canine cadaver femurs, and biomechanical properties were measured, including peak insertional torque, torque to failure, and pullout strength, each normalized to cortical thickness. Scanning electron micrographs were taken from 24 screws, and images were blindly analyzed by 5 trained examiners. RESULTS: The mean normalized insertion torque for 3.5-mm screws was significantly different between G1 and both G50 and G100. The mean normalized torque to failure for 3.5-mm screws was significantly different between G1 and both G50 and G100. Axial pullout testing was found to be significantly different for 2.0-mm screws between G1 and G100. Scanning electron micrographs surface scoring identified a significant difference in 3.5-mm screws at the screw tip. CONCLUSIONS: The results indicate that biomechanical changes occur with repeated steam sterilizations. Specifically, peak insertional torque and torque to failure are decreased with increased sterilizations for 3.5-mm screws, whereas 2.0-mm screws were altered in pullout testing after 100 sterilizations. It is suspected that numerous sterilizations negatively alter the physical-mechanical properties of certain screw sizes. CLINICAL RELEVANCE: The biomechanical properties of the bone-implant interface could negatively be affected by multiple steam sterilizations during clinical setting.

The influence of a tension band fixation as an adjunct for arthrodesis of the metacarpophalangeal joint in the horse.

OBJECTIVES: To determine the influence of a stainless-steel cable (SSC) tension band fixation as an adjunct to a locking compression plate (LCP) for arthrodesis of the equine metacarpophalangeal (MCP) joint. STUDY DESIGN: Experimental. An ex vivo biomechanical paired equine cadaver limb study. SAMPLE POPULATION: Five MCP joint pairs were collected from adult Thoroughbred horses, euthanized for reasons unrelated to orthopedic disease. METHODS: Each pair of MCP joints were randomly implanted with either a dorsally placed 5.5 mm LCP and a palmarly placed 2.0 mm SSC or a dorsally placed 5.5 mm LCP alone. Each construct was tested in cyclic loading followed by single cycle to failure in axial compression. Displacement at a target load of 1 kN over 3600 cycles at 1 Hz was recorded prior to single cycle to failure testing. RESULTS: In cyclic testing, displacement was not significantly different between the first and last 5% of testing cycles regardless of construct. Maximum displacement of each construct during cyclic testing was <1.1 mm. In single cycle testing, the observed yield point did not reveal any difference between LCP and LCP-SSC (p = .440). The maximum load at failure was significantly higher in LCP-SSC compared to constructs with the LCP alone (p = .046). CONCLUSION: The addition of the SSC to the LCP did not statistically affect construct displacement during cyclic loading or construct yield load during subsequent single cycle to failure. CLINICAL SIGNIFICANCE: This study provided much needed information regarding the necessity of a tension band SSC application in the arthrodesis of the MCP/MTP joint in horses.

DYNAMIC SONOGRAPHY OF THE EQUINE METACARPO(TARSO)PHALANGEAL DIGITAL FLEXOR TENDON SHEATH.

Palmar/plantar annular desmitis is a common disease that may be associated with adhesions and structures affecting the flexor tendons, which requires tenoscopy to diagnose. The purpose of this descriptive study was to develop a dynamic sonographic technique for evaluating the motion of normal equine flexor tendons in relation to the palmar/plantar annular ligament and to compare findings with horses previously diagnosed with palmar/plantar annular desmitis. Ten healthy adult horses were examined prospectively and the images of four horses diagnosed with palmar/plantar annular desmitis were retrospectively evaluated. Dynamic sonography was performed at the level of the metacarpo/metatarsophalangeal joint by maximally extending and flexing the interphalangeal joints. Palmar/plantar annular ligament thickness (mm), size of any gap between the flexor tendons, and subjectively increased angulation of the long linear echoes of the superficial digital flexor tendon were measured. The presence of gliding motion between the palmar/plantar annular ligament and superficial digital flexor tendon was determined by consensus. Twenty-eight healthy control limbs (16 hind/12 fore) and four with palmar/plantar annular desmitis (3 hind/1 fore) were evaluated. Controls had unrestricted gliding motion between the palmar/plantar annular ligament and flexor tendons and zero to 13° of angulation of the long linear echoes. The four affected horses had restricted gliding motion and between 20-35° angulation of the long linear echoes. Dynamic ultrasound is a feasible technique for detecting restricted flexor tendon and palmar/plantar annular ligament gliding motion, as well as subjectively increased angulation of the long linear echoes of the flexor tendon in affected horses compared with controls and warrants further investigation.

An in vitro biomechanical comparison of hydroxyapatite coated and uncoated ao cortical bone screws for a limited contact: dynamic compression plate fixation of osteotomized equine 3rd metacarpal bones.

OBJECTIVES: To compare the monotonic biomechanical properties of a broad 4.5 mm limited contact-dynamic compression plate (LC-DCP) fixation secured with hydroxyapatite (HA) coated cortical bone screws (HA-LC-DCP) versus uncoated cortical bone screws (AO-LC-DCP) to repair osteotomized equine 3rd metacarpal (MC3) bones. STUDY DESIGN: Experimental. SAMPLE POPULATION: Adult equine cadaveric MC3 bones (n = 12 pair). METHODS: Twelve pairs of equine MC3 were divided into 3 test groups (4 pairs each) for: (1) 4 point bending single cycle to failure testing; (2) 4 point bending cyclic fatigue testing; and (3) torsional single cycle to failure testing. For the HA-LC-DCP-MC3 construct, an 8-hole broad LC-DCP (Synthes Ltd, Paoli, PA) was secured on the dorsal surface of each randomly selected MC3 bone with a combination of four 5.5 mm and four 4.5 mm HA-coated cortical screws. For the AO-LC-DCP-MC3 construct, an 8-hole 4.5 mm broad LC-DCP was secured on the dorsal surface of the contralateral MC3 bone with a combination of four 5.5 mm and four 4.5 mm uncoated cortical screws. All MC3 bones had mid-diaphyseal osteotomies. Mean test variable values for each method were compared using a paired t-test within each group. Significance was set at P < .05. RESULTS: Mean yield load, yield bending moment, composite rigidity, failure load, and failure bending moment, under 4 point bending, single cycle to failure, of the HA-LC-DCP fixation were significantly greater than those of the AO-LC-DCP fixation. Mean ± SD values for the HA-LC-DCP and the AO-LC-DCP fixation techniques, respectively, in single cycle to failure under 4 point bending were: yield load, 26.7 ± 2.15 and 16.3 ± 1.38 kN; yield bending moment, 527.4 ± 42.4 and 322.9 ± 27.2 N-m; composite rigidity, 5306 ± 399 and 3003 ± 300 N-m/rad; failure load, 40.6 ± 3.94 and 26.5 ± 2.52 kN; and failure bending moment, 801.9 ± 77.9 and 522.9 ± 52.2 N-m. Mean cycles to failure in 4 point bending of the HA-LC-DCP fixation (116,274 ± 13,211) was significantly greater than that of the AO-LC-DCP fixation 47,619 ± 6580. Mean yield load, mean composite rigidity, and mean failure load under torsional testing, single cycle to failure was significantly greater for the broad HA-LC-DCP fixation compared with the AO-LC-DCP fixation. In single cycle to failure under torsion, mean ± SD values for the HA-LC-DCP and the AO-LC-DCP fixation techniques, respectively, were: yield load, 101.3 ± 14.68 and 70.54 ± 10.20 N-m; composite rigidity, 437.9 ± 32.9 and 220.7 ± 17.6 N-m/rad; and failure load: 105.7 ± 15.5 and 75.28 ± 10.1 N-m. CONCLUSION: HA-LC-DCP was superior to AO-LC-DCP in resisting the static overload forces (palmarodorsal 4 point bending and torsional) and in resisting cyclic fatigue under palmarodorsal 4 point bending.

Long term outcome after laser assisted modified Forssell's in cribbing horses.

OBJECTIVE: To (1) report long-term outcome after laser assisted modified Forssell's procedure; (2) describe the prevalence of postoperative complications; and (3) identify risk factors associated with outcome. STUDY DESIGN: Retrospective case series. ANIMALS: Horses (n = 119). METHODS: Medical records (1994-2012) of horses that had laser assisted modified Forssell's procedure were reviewed. Signalment, preoperative duration of cribbing, postoperative complications and outcome were recorded. RESULTS: Follow-up was available for 90 horses; 76 had stopped cribbing for >1 year. Of 14 horses that resumed cribbing, median relapse time was 6 months. Preoperative cribbing duration was significantly associated with an increased probability of unsuccessful outcome when horses cribbed for >3 years before surgery (odds ratio of positive outcome: OR = 0.14, 95% CI: 0.02, 0.95). Overall, 22.2% horses had postoperative complications (hematoma, seroma, infection, and prolonged drainage). CONCLUSIONS: Laser assisted modified Forssell's is a very effective (84.4%) surgical treatment of cribbing behavior in horses. Preoperative cribbing duration was identified as a significant risk factor associated with unsuccessful surgical outcome.

Magnetic resonance imaging in foals with infectious arthritis.

The magnetic resonance (MR) imaging findings of foals with infectious and noninfectious arthritis are described. Six foals with infectious arthritis and three foals with noninfectious arthritis were grouped based on synovial fluid analysis results and examined with radiography and MR imaging. Four out of six foals with infectious arthritis had osseous lesions in MR images indicative of osteomyelitis and only 4/19 lesions were detected on digital radiographs. The three foals with noninfectious arthritis had no osseous lesions in MR images or radiographically. Of the six joints that had osseous lesions detected with MR imaging, three had at least one lytic lesion detected radiographically. Osseous lesions in the epiphysis, metaphysis, and physis appeared in MR images as T2W, short tau inversion recovery, and proton density hyperintense foci with a hypointense halo. The same lesions appeared hyperintense in the 3D RSSG water excitation pulse sequence but lacked a surrounding hypointense halo. Most joints of foals with infectious arthritis had heterogenous signals within the synovial fluid whereas all of the nonseptic joints had homogenous synovial fluid signals. MR imaging appears to be better than radiography in the detection of osseous lesions in foals diagnosed with infectious arthritis and may be a valuable screening test for the presence of osteomyelitis.

In vitro biomechanical comparison of dynamic compression plates with a rough contact surface and a polished contact surface for fixation of osteotomized equine third metacarpal bones.

OBJECTIVES: To compare the number of cycles to failure of 4.5 mm broad dynamic compression plates (DCP), 4.5 mm broad limited-contact dynamic compression plates (4.5-LC-DCP), and 5.5 mm broad limited-contact dynamic compression plates (5.5-LC-DCP) having a rough (denoted by a prefix R-) versus a standard smooth contact surface for the fixation of osteotomized equine 3rd metacarpal (MC3) bones. STUDY DESIGN: Experimental. ANIMAL POPULATION: Fifteen pairs of adult equine cadaveric MC3 bones. METHODS: Fifteen pairs of equine MC3 were divided into 3 test groups (5 pairs each) for comparison of (1) R-DCP fixation with DCP fixation, (2) R-4.5-LC-DCP fixation with 4.5-LC-DCP fixation, and (3) R-5.5-LC-DCP fixation with 5.5-LC-DCP fixation to repair osteotomized equine MC3 bones under palmarodorsal 4-point bending cyclic fatigue testing. For each group an 8-hole plate with rough contact surface was applied to the dorsal surface of one randomly selected bone from each pair and a corresponding 8-hole plate with smooth contact surface was applied dorsally to the contralateral bone from each pair. All plates and screws were applied using standard ASIF techniques. All MC3 bones had mid-diaphyseal osteotomies. Mean number of cycles to failure for each method were compared using a paired t-test within each group. Significance was set at P < .05. RESULTS: Mean cycles to failure ± standard deviation was significantly greater for the R-DCP fixation (230,025 ± 23,129) compared with the DCP fixation (103,451 ± 14,556), for the R-4.5-LC-DCP fixation (99,237 ± 14,390) compared with the 4.5-LC-DCP fixation (46,464 ± 6325) and for the R-5.5-LC-DCP fixation (65,113 ± 7796) compared with the 5.5-LC-DCP fixation (34,224 ± 3835). CONCLUSION: For the fixation of osteotomized MC3 bones, the constructs with plates having rough contact surface were superior to the corresponding constructs with plates having standard smooth contact surfaces in resisting cyclic fatigue under palmarodorsal 4-point bending.

A mechanical comparison of equine proximal interphalangeal joint arthrodesis techniques: an axial locking compression plate and two abaxial transarticular cortical screws versus an axial dynamic compression plate and two abaxial transarticular cortical screws.

OBJECTIVES: To compare in vitro monotonic biomechanical properties of an axial 3-hole, 4.5 mm narrow locking compression plate (ELCP) using 5.0 mm locking screws and 5.5 mm cortical screws in conjunction with 2 abaxial transarticular 5.5 mm cortical screws inserted in lag fashion (ELCP-TLS) with an axial 3-hole, 4.5 mm narrow dynamic compression plate (DCP) using 5.5 mm cortical screws in conjunction with 2 abaxial transarticular 5.5 mm cortical screws inserted in lag fashion (DCP-TLS) for equine proximal interphalangeal (PIP) joint arthrodesis. DESIGN: Experimental. ANIMAL POPULATION: Cadaveric adult equine forelimbs (n = 18 pairs). METHODS: For each forelimb pair, 1 PIP joint was stabilized with an axial ELCP using 5.0 mm locking screws and 5.5 mm cortical screws in conjunction with 2 abaxial transarticular 5.5 mm cortical screws inserted in lag fashion and 1 PIP joint with an axial 3-hole narrow DCP (4.5 mm) using 5.5 mm cortical screws in conjunction with 2 abaxial transarticular 5.5 mm cortical screws inserted in lag fashion. Six matching pairs of constructs were tested in single cycle to failure under axial compression, 6 construct pairs were tested for cyclic fatigue under axial compression, and 6 construct pairs were tested in single cycle to failure under torsional loading. Mean values for each fixation method were compared using a paired t-test within each group with statistical significance set at P < .05. RESULTS: Mean yield load, yield stiffness, and failure load under axial compression, single cycle to failure, of the DCP-TLS fixation were significantly greater than those of the LCP-TLS fixation. There was no significant difference between the mean number of cycles to failure in axial compression of the LCP-TLS and the DCP-TLS fixations. Mean yield load, yield stiffness, and failure load under torsion, single cycle to failure, of the LCP-TLS fixation were significantly greater than those of the DCP-TLS fixation. CONCLUSION: The DCP-TLS construct provided significantly greater stability under axial compression in single cycle to failure than the ELCP-TLS construct, the ELCP-TLS construct provided significantly greater stability under torsional loading in single cycle to failure than the DCP-TLS construct, and there was no significant difference in stability between the 2 constructs for cyclic loading under axial compression.

An in vitro biomechanical comparison of equine proximal interphalangeal joint arthrodesis techniques: two parallel transarticular headless tapered variable pitch screws versus two parallel transarticular AO cortical bone screws inserted in lag fashion.

OBJECTIVES: To compare the mean number of cycles to failure under axial compression of equine proximal interphalangeal (PIP) joint arthrodesis constructs created by 2 parallel transarticular Acutrak Plus screws (AP-TS) or 2 parallel transarticular 5.5 mm cortical screws inserted in lag fashion (AO-TLS). STUDY DESIGN: Paired in vitro biomechanical testing of 2 methods of stabilizing cadaveric adult equine forelimb PIP joints. SAMPLE POPULATION: Cadaveric adult equine forelimbs (n=5 pairs). METHODS: For each forelimb pair, 1 PIP joint was stabilized with AP-TS and 1 with AP-TLS. The 5 construct pairs were tested for cyclic fatigue under axial compression. Mean number of cycles to failure for each fixation method were compared by a paired t-test within each group with statistical significance set at P<.05. RESULTS: The mean number of cycles to failure under axial compression for AO-TLS fixation and AP-TS fixation were 57,723±8488 and 35,322±4698, respectively. CONCLUSION: The AO-TLS was superior to the AP-TS in resisting cyclic fatigue under axial compression.

In vitro biomechanical comparison of a modified 5.5 mm locking compression plate fixation with a 5.5 mm locking compression plate fixation of osteotomized equine third metacarpal bones.

OBJECTIVES: To compare number of cycles to failure for palmarodorsal 4-point bending of a modified 5.5 mm broad locking compression plate (M5.5-LCP) fixation with a 5.5 mm broad LCP (5.5-LCP) fixation used to repair osteotomized equine third metacarpal (MC3) bones. STUDY DESIGN: In vitro biomechanical testing. ANIMAL POPULATION: Adult equine cadaveric MC3 bones (n=6 pairs). METHODS: An 8-hole, M5.5-LCP, obtained by having a 1.0 mm thickness removed from the bone contact portion of the 5.5-LCP, was applied to the dorsal surface of 1 randomly selected MC3 from each pair, and an 8-hole, 5.5-LCP was applied dorsally to the contralateral bone from each pair using a combination of cortical and locking screws. Plates and screws were applied using standard ASIF techniques to MC3 bones with a mid-diaphyseal osteotomy. MC3 constructs had palmarodorsal 4-point bending cyclic fatigue testing. Mean cycles to failure for each method were compared using a paired t-test within each group. Significance was set at P<.05. RESULTS: Mean±SD cycles to failure of the M5.5-LCP fixation (188,641±17,971) was significantly greater than that of the 5.5-LCP fixation (166,497±15,539). CONCLUSION: M5.5-LCP fixation was superior to 5.5-LCP fixation of osteotomized equine MC3 bones in resisting cyclic fatigue under palmarodorsal 4-point bending. CLINICAL RELEVANCE: This suggests that biological plate fixation is not the ideal choice for osteotomized equine MC3 bones.

An in vitro biomechanical comparison of a 5.5 mm locking compression plate fixation with a 4.5 mm locking compression plate fixation of osteotomized equine third metacarpal bones.

OBJECTIVES: To compare the monotonic biomechanical properties and fatigue life of a 5.5-mm-broad locking compression plate (5.5 LCP) fixation with a 4.5-mm-broad locking compression plate (4.5 LCP) fixation to repair osteotomized equine 3rd metacarpal (MC3) bones. STUDY DESIGN: In vitro biomechanical testing of paired cadaveric equine MC3 with a middiaphyseal osteotomy, stabilized by 1 of 2 methods for fracture fixation. ANIMAL POPULATION: Fifteen pairs of adult equine cadaveric MC3 bones. METHODS: Fifteen pairs of equine MC3 were divided into 3 test groups (5 pairs each) for (1) 4-point bending single cycle to failure testing, (2) 4-point bending cyclic fatigue testing, and (3) torsional single cycle to failure testing. An 8-hole, 5.5 LCP was applied to the dorsal surface of 1 randomly selected bone from each pair and an 8-hole, 4.5 LCP was applied dorsally to the contralateral bone from each pair using a combination of cortical and locking screws. All plates and screws were applied using standard ASIF techniques. All MC3 bones had middiaphyseal osteotomies. Mean test variable values for each method were compared using a paired t-test within each group with significance set at P<.05. RESULTS: Mean yield load, yield bending moment, composite rigidity, failure load, and failure bending moment, under 4-point bending, single cycle to failure, of the 5.5 LCP fixation were significantly greater than those of the 4.5 LCP fixation. Mean cycles to failure in 4-point bending of the 5.5 LCP fixation (170,535+/-19,166) was significantly greater than that of the 4.5 LCP fixation (129,629+/-14,054). Mean yield load, mean composite rigidity, and mean failure load under torsional testing, single cycle to failure was significantly greater for the broad 5.5 LCP fixation compared with the 4.5 LCP fixation. In single cycle to failure under torsion, the mean+/-SD values for the 5.5 LCP and the 4.5 LCP fixation techniques, respectively, were: yield load, 151.4+/-19.6 and 97.6+/-12.1 N m; composite rigidity, 790.3+/-58.1 and 412.3+/-28.1 N m/rad; and failure load: 162.1+/-20.2 and 117.9+/-14.6 N m. CONCLUSION: The 5.5 LCP was superior to the 4.5 LCP in resisting static overload forces (palmarodorsal 4-point bending and torsional) and in resisting cyclic fatigue under palmarodorsal 4-point bending. CLINICAL RELEVANCE: These in vitro study results may provide information to aid in selection of an LCP for repair of equine long bone fractures.

An in vitro biomechanical comparison of equine proximal interphalangeal joint arthrodesis techniques: an axial positioned dynamic compression plate and two abaxial transarticular cortical screws inserted in lag fashion versus three parallel transarticular cortical screws inserted in lag fashion.

OBJECTIVES: To compare in vitro monotonic biomechanical properties of an axial 3-hole, 4.5 mm narrow dynamic compression plate (DCP) using 5.5 mm cortical screws in conjunction with 2 abaxial transarticular 5.5 mm cortical screws inserted in lag fashion (DCP-TLS) with 3 parallel transarticular 5.5 mm cortical screws inserted in lag fashion (3-TLS) for the equine proximal interphalangeal (PIP) joint arthrodesis. STUDY DESIGN: Paired in vitro biomechanical testing of 2 methods of stabilizing cadaveric adult equine forelimb PIP joints. SAMPLE POPULATION: Cadaveric adult equine forelimbs (n=15 pairs). METHODS: For each forelimb pair, 1 PIP joint was stabilized with an axial 3-hole narrow DCP (4.5 mm) using 5.5 mm cortical screws in conjunction with 2 abaxial transarticular 5.5 mm cortical screws inserted in lag fashion and 1 with 3 parallel transarticular 5.5 mm cortical screws inserted in lag fashion. Five matching pairs of constructs were tested in single cycle to failure under axial compression, 5 construct pairs were tested for cyclic fatigue under axial compression, and 5 construct pairs were tested in single cycle to failure under torsional loading. Mean values for each fixation method were compared using a paired t-test within each group with statistical significance set at P<.05. RESULTS: Mean yield load, yield stiffness, and failure load under axial compression and torsion, single cycle to failure, of the DCP-TLS fixation were significantly greater than those of the 3-TLS fixation. Mean cycles to failure in axial compression of the DCP-TLS fixation was significantly greater than that of the 3-TLS fixation. CONCLUSION: The DCP-TLS was superior to the 3-TLS in resisting the static overload forces and in resisting cyclic fatigue. CLINICAL RELEVANCE: The results of this in vitro study may provide information to aid in the selection of a treatment modality for arthrodesis of the equine PIP joint.

Equine neutrophil elastase in plasma, laminar tissue, and skin of horses administered black walnut heartwood extract.

Laminitis is a local manifestation of a systemic inflammatory response that is characterized by neutrophil activation and movement of neutrophils into the laminar tissues. Given the evidence for the involvement of neutrophils in the development of laminitis, we measured concentrations of neutrophil elastase, a serine protease released from the azurophilic granules of neutrophils, in plasma, skin and laminar tissues obtained from control horses and horses given black walnut heartwood extract (BWHE) to induce laminitis. Healthy horses (5-15 years old) were randomly assigned to 4 groups: 3 experimental groups given BWHE via nasogastric tube, and a control group given an equal volume of water. The experimental groups consisted of horses euthanized 1.5h (n=5), 3h (n=6) or 12h (n=10) after BWHE administration. Control horses (n=7) were euthanized 12h after intragastric administration of water. Plasma samples were collected in all horses of the control and 12h BWHE groups at 0, 1, 2, 3, 4, 6, 8, 10, and 12h after treatment, and laminar tissue and skin from the middle region of the neck were harvested at the time of euthanasia in all 1.5 and 3h BWHE horses, in 6 of the 12h BWHE horses and in 5 of the control horses. Plasma and tissue concentrations of neutrophil elastase were determined using an equine specific ELISA, and statistical significance was set at p<0.05. Plasma concentrations of neutrophil elastase in the BWHE group were significantly higher at 6 and 8h compared to the control group and at 8 and 10h compared to time 0. Concentrations of neutrophil elastase in skin and laminar tissue were significantly higher in the 3 and 12h BWHE groups compared to the control group. Concentrations of neutrophil elastase were significantly higher in the skin than in the lamina in the 12h BWHE horses. The administration of BWHE thus results in significant increases in the concentration of neutrophil elastase in the circulation, skin and laminar tissue. These results confirm a role for neutrophils in the developmental phase of laminitis, and the systemic nature of the inflammatory process. Furthermore, neutrophil elastase may play a key role in the disintegration of the hoof basal membrane and be a target for the development of new treatments for laminitis.

Comparison of characteristics and enzymatic products of leukocytes in the skin and laminar tissues of horses administered black walnut heartwood extract or lipopolysaccharide.

OBJECTIVE: To compare characteristics and enzymatic products of leukocytes detected in the skin and laminar tissues of horses administered black walnut heartwood extract (BWHE) and horses administered purified lipopolysaccharide (LPS). ANIMALS: 25 healthy 5- to 15-year-old horses. PROCEDURES: Horses were randomly assigned to receive LPS (20 ng of O55:B5 Escherichia coli endotoxin/kg; n = 5) IV or 6 L of BWHE (10) or water (control group; 10) via nasogastric intubation. Horses were euthanatized 12 hours after treatment or at onset of Obel grade 1 lameness. Laminar tissue samples and skin samples from the middle region of the neck were harvested at the time of euthanasia. Leukocyte emigration (determined via CD13 immunohistochemical analysis) and matrix metalloproteinase (MMP)-2 and MMP-9 gene expressions and activities (determined via reverse transcription PCR assay and gelatin zymography, respectively) were measured in skin and laminar tissue samples. RESULTS: Tissues of horses receiving BWHE contained significantly higher numbers of CD13-positive cells and increased MMP-9 gene expression and activity, compared with findings in the other 2 groups. Values for laminar tissue and skin from LPS-treated horses were not increased, compared with findings in the control group, in any experiment. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that BWHE administration causes increases in CD13-positive leukocyte numbers and MMP-9 expression and activity in laminar tissue and skin in horses; similar effects were not detected following LPS administration. Leukocyte emigration in horses with experimentally induced endotoxemia and in horses administered BWHE differed markedly, thereby providing additional evidence that the development of laminitis involves more complex mechanisms than endotoxemia-induced leukocyte activation alone.

Neutrophil myeloperoxidase measurements in plasma, laminar tissue, and skin of horses given black walnut extract.

OBJECTIVE: To compare measurements of myeloperoxidase (MPO) in plasma, laminar tissues, and skin obtained from control horses and horses given black walnut heartwood extract (BWHE). ANIMALS: 22 healthy 5- to 15-year-old horses. PROCEDURES: Horses were randomly assigned to 4 groups as follows: a control group given water (n = 5) and 3 experimental groups given BWHE (17) via nasogastric intubation. Experimental groups consisted of 5, 6, and 6 horses that received BWHE and were euthanatized at 1.5, 3, and 12 hours after intubation, respectively. Control horses were euthanatized at 12 hours after intubation. Plasma samples were obtained hourly for all horses. Laminar tissue and skin from the middle region of the neck were harvested at the time of euthanasia. Plasma and tissue MPO concentrations were determined via an ELISA; tissue MPO activity was measured by use of specific immunologic extraction followed by enzymatic detection. RESULTS: Tissues and plasma of horses receiving BWHE contained significantly higher concentrations of MPO beginning at hour 3. Laminar tissue and skin from horses in experimental groups contained significantly higher MPO activity than tissues from control horses. Concentrations and activities of MPO in skin and laminar tissues were similar over time. CONCLUSIONS AND CLINICAL RELEVANCE: In horses, BWHE administration causes increases in MPO concentration and activity in laminar tissue and skin and the time of increased MPO concentration correlates with emigration of WBCs from the vasculature. These findings support the hypothesis that activation of peripheral WBCs is an early step in the pathogenesis of acute laminitis.