Laura M. Riggs

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.

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.

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.

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.

Long Term Outcome After Laser Assisted Modified Forssell's in Cribbing Horses

Objective To (1) report long-term outcome after laser assisted modified Forssells 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 Forssells 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 Forssells 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.OBJECTIVE To (1) report long-term outcome after laser assisted modified Forssells 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 Forssells 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 Forssells 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.

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.

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 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.

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.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.