Outcomes of podotrochlear (navicular) bursa injections for signs of foot pain in horses evaluated via magnetic resonance imaging: 23 cases (2005-2007).

OBJECTIVE: To determine clinical outcome following intrathecal injection of the podotrochlear (navicular) bursa for signs of foot pain in horses evaluated via magnetic resonance imaging (MRI) and evaluate efficacy of corticosteroids administered with or without hyaluronate. DESIGN: Retrospective case series. ANIMALS: 23 horses. PROCEDURES: Data collected included signalment, history, intended use, duration and severity of lameness, results of diagnostic anesthesia, radiographic abnormalities, MRI abnormalities, and outcomes for return to use. RESULTS: MRI was conducted on 23 horses with lameness localized to the foot. Thirteen horses had bilateral forelimb lameness, and 10 had unilateral forelimb lameness. Mean duration of lameness was 10.5 months. Seventeen of 23 (74%) horses had excellent outcomes and returned to intended use within 2 to 4 weeks after navicular bursa injection. Hyaluronate treatment was not associated with outcome; however, horses receiving < 10 mg of trimacinolone had significantly worse outcomes than those treated with hyaluronate. Among horses with excellent outcomes, mean duration of soundness was 7.3 months. Seven of 8 horses with erosive lesions of the flexor surface of the distal sesamoid (navicular) bone diagnosed via MRI had a poor outcome. Horses with navicular bursitis responded optimally to injection, compared with horses with other problems. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that intrathecal injection of corticosteroid in horses with erosions of the flexor surface of the navicular bone associated with deep digital flexor tendon adhesions yielded a poor response. Treatment of horses with navicular bursitis via injection of the navicular bursa should be highly effective in alleviating lameness.

Alginate encapsulation impacts the insulin-like growth factor-I system of monolayer-expanded equine articular chondrocytes and cell response to interleukin-1beta.

Alginate hydrogel culture has been shown to reestablish chondrocytic phenotype following monolayer expansion; however, previous studies have not adequately addressed how culture conditions affect the signaling systems responsible for chondrocyte metabolic activity. Here we investigate whether chondrocyte culture history influences the insulin-like growth factor-I (IGF-I) signaling system and its regulation by interleukin-1 (IL-1). Articular chondrocytes (ACs) from equine stifle joints were expanded by serial passage and were either encapsulated in alginate beads or maintained in monolayer culture for 10 days. Alginate-derived cells (ADCs) and monolayer-derived cells (MDCs) were then plated at high density, stimulated with IL-1beta (1 and 10 ng/mL) or IGF-I (50 ng/mL) for 48 h, and assayed for levels of type I IGF receptor (IGF-IR), IGF binding proteins (IGFBPs), and endogenously secreted IGF-I. Intermediate alginate culture yielded relatively low IGF-IR levels that increased in response to IL-1beta, whereas higher receptor levels on MDCs were reduced by cytokine. MDCs also secreted substantially more IGFBP-2, the predominant binding protein in conditioned media (CM), though IL-1beta suppressed levels for both cell populations. Concentrations of autocrine/paracrine IGF-I paralleled IGFBP-2 secretion. Disparate basal levels of IGF-IR and IGFBP-2, but not IGF-I, were attributed to relative transcript expression. Systemic differences coincided with varied effects of IL-1beta and IGF-I on cell growth and type I collagen expression. We conclude that culture strategy impacts the IGF-I signaling system of ACs, potentially altering their capacity to mediate cartilage repair. Consideration of hormonal regulators may be an essential element to improve chondrocyte culture protocols used in tissue engineering applications.

Transcriptional and proteolytic regulation of the insulin-like growth factor-I system of equine articular chondrocytes by recombinant equine interleukin-1beta.

Interleukin-1 (IL-1) and insulin-like growth factor-I (IGF-I), which have opposing effects on matrix metabolism within articular cartilage, are thought to play prominent roles in the pathogenesis of osteoarthritis. To better understand the link between these anabolic (IGF-I) and catabolic (IL-1) stimuli, we examined exogenous IL-1 regulation of the IGF-I signaling system of articular chondrocytes (ACs). Equine ACs from non-arthritic stifle joints were expanded in monolayer culture, encapsulated for 10 days in alginate beads, and stimulated as high-density monolayers with recombinant equine IL-1beta (0, 1, 10 ng/ml) for 48 h. IL-1beta enhanced expression of IGF-IR levels, as determined by both [125I]-IGF-I binding studies and Western blotting, while reducing the concentration of endogenous IGF-I detected in conditioned media by radioimmunoassay. Western ligand blotting revealed that chondrocytes primarily secreted IGF binding proteins (IGFBPs) with molecular weights of 28-30 and 32-34 kDa, which were identified as IGFBPs 5 and 2, respectively, and that IL-1beta treatment diminished IGFBP-2, the prominent homolog in conditioned media. Northern blot analysis suggested IL-1beta regulation of IGF-I and, to some extent, IGF-IR was mediated by transcription; however, the cytokine did not affect IGFBP-2 expression. To test for evidence of proteolysis by matrix metalloproteinases (MMPs), additional cultures were co-incubated with inhibitors for MMPs 2/9, 3, and 8. IGFBP-2 suppression was partially reversed by gelatinase (MMP-2/9) inhibition. In summary, these findings further delineate the role of IL-1 as a key regulator of the IGF-I system within articular cartilage, demonstrating that regulation occurs through both direct (transcriptional) and indirect (proteolytic) mechanisms.

Hand-assisted laparoscopic ovariohysterectomy in the mare.

OBJECTIVE: To develop a minimally invasive, hand-assisted laparoscopic ovariohysterectomy (HALS-OHE) technique in the mare and to evaluate safety and any associated complications. STUDY DESIGN: Experimental study. ANIMALS: Eight, 11-24-year-old mares with anatomically normal urogenital tracts. METHODS: The surgical technique was developed in 2 non-survival mares and subsequently evaluated in 6 survival procedures. Food was withheld for 48 hours, then mares were anesthetized and positioned in dorsal recumbency for laparoscopic surgery. A hand access device (Omniport) was placed followed by 4 laparoscopic portals. Transection of the ovarian pedicles and broad ligaments was achieved using a combination of a laparoscopic stapling instrument (Endo GIA II), an ultrasonically activated instrument (Harmonic Scalpel), and endoscopic clips (Endo Clip II ML). The genital tract was exteriorized through the laparotomy, and the uterus transected and sutured in a conventional pattern. Horses were evaluated through postoperative day 14 when a necropsy was performed. RESULTS: Four mares recuperated well after surgery, 1 mare was euthanatized because of bilateral femur fracture during anesthetic recovery, and another developed severe pleuropneumonia. At necropsy all but 1 abdominal incision was healing routinely. One mare had abscessed along the celiotomy incision and developed visceral adhesions. Uncomplicated healing of transected mesovarial, mesometrial, and uterine remnants was observed. CONCLUSIONS: Ovariohysterectomy in horses can be accomplished using HALS technique. CLINICAL RELEVANCE: HALS-OHE technique represents a minimally invasive and technically feasible alternative for conventional OHE. Careful patient selection and preparation may reduce the complications observed. The HALS technique may be useful in other laparoscopic surgical procedures.

Elution of metronidazole and gentamicin from polymethylmethacrylate beads.

OBJECTIVE: To characterize the elution and bioactivity of metronidazole and gentamicin sulfate polymerized, individually and in combination, with polymethylmethacrylate (PMMA). STUDY DESIGN: In vitro experimental study. METHODS: PMMA beads containing metronidazole (3 concentrations), gentamicin sulfate, or metronidazole and gentamicin sulfate were immersed in 5 mL of phosphate-buffered saline in triplicate. Eluent was replaced at specified time intervals for 1 or 21 days, and antibiotic concentrations were measured by high-performance liquid chromatography. Changes in antibiotic bioactivity attributable to polymerization or copolymerization of the antibiotics with PMMA, ethylene oxide sterilization, and storage of AIPMMA beads containing metronidazole were evaluated. RESULTS: Antibiotic elution patterns were similar for all groups. Day 1 elution for groups containing metronidazole or gentamicin individually represented a mean 63%-66% and 79%, respectively, of the 21-day total. Approximately 50% of the day 1 elution occurred during the first hour. The elution of metronidazole was dose dependent. The elution of metronidazole (day 3-21) and gentamicin (all days) was significantly greater when metronidazole and gentamicin were combined (P <.05). The addition of metronidazole delayed polymerization of PMMA. Neither polymerization nor copolymerization of metronidazole and gentamicin with PMMA, gas sterilization, or 2-month storage of beads containing metronidazole significantly affected antimicrobial bioactivity. CONCLUSIONS: Metronidazole elution from PMMA was dose dependent. Copolymerization of metronidazole and gentamicin sulfate in PMMA resulted in increased rates of elution. Intraoperative preparation of metronidazole-impregnated PMMA beads is not practical, but sterilization and storage for 2 months should not affect efficacy. CLINICAL RELEVANCE: The local delivery of biologically active metronidazole and gentamicin by elution from PMMA is feasible.

Evaluation of the harmonic scalpel for laparoscopic bilateral ovariectomy in standing horses.

OBJECTIVE: To evaluate use of the Harmonic Scalpel (Ethicon Endo-Surgery Inc., Cincinnati, OH) for performing laparoscopic bilateral ovariectomy in standing horses. STUDY DESIGN: Experimental study. ANIMALS: Eight mares aged 2-20 years and weighing 410-540 kg. METHODS: Standing laparoscopic bilateral ovariectomy was performed in 8 mares with normal reproductive tract anatomy. The Harmonic Scalpel (an ultrasonically activated instrument) was used to simultaneously transect and obtain hemostasis of the ovarian pedicle. Necropsy was performed on 4 mares 3 days after surgery and 4 mares 30 days after surgery. Gross and histopathologic evaluation of the ovarian pedicles was performed to characterize tissue reaction. RESULTS: Complete hemostasis of the ovarian pedicles was obtained in all mares. Median transection time for the ovarian pedicle was 28 minutes. Postoperative complications included transient fever, moderate subcutaneous emphysema, and incisional seroma formation. On necropsy examination, there were no signs of generalized peritonitis, postoperative hemorrhage, or adhesion formation. Mild to moderate acute inflammation and scar formation with moderate chronic inflammation at the ovarian pedicle was found at 3 and 30 days. Median depth of coagulation necrosis at 3 days was 2.87 mm. CONCLUSIONS: The Harmonic Scalpel appears to provide reliable hemostasis of the ovarian pedicle during elective laparoscopic ovariectomy in horses. Clinical Relevance-The Harmonic Scalpel represents a safe alternative to other methods of hemostasis during elective laparoscopic ovariectomy in horses.

Experimental evaluation of urinary bladder marsupialization in male goats.

OBJECTIVE: To evaluate the outcome of urinary bladder marsupialization in male goats. STUDY DESIGN: Prospective, experimental study. ANIMALS: Six healthy mixed-breed male goats. METHODS: After experimentally induced urethral obstruction, 6 male goats had urinary bladder marsupialization. Renal ultrasonography, complete blood count, and serum biochemical analysis were evaluated preoperatively (day 0), at 7 postoperative days, and then at 30-day intervals until 180 days. Stomal diameter was recorded immediately postoperatively and at each postoperative interval. Necropsy examination was performed on day 180 or when stomal stricture or death occurred. RESULTS: Stomal stricture occurred in 1 goat at 120 days. Another goat was found dead at 150 days; severe, suppurative cystitis was identified on necropsy. All goats had mild urine scald dermatitis. Serum biochemical values remained within normal limits, but significant decreases in white blood cell count, serum creatinine concentration, and stomal diameter occurred. At necropsy, all bladders were tubular in shape. Histological evidence of chronic suppurative cystitis and chronic, mild lymphoplasmacytic pyelitis occurred in all goats. Bacterial culture of renal tissue yielded growth in 3 goats, and bladder mucosal swabs yielded bacterial growth in all goats. CONCLUSIONS: Although clinical signs of ascending urinary tract infection were not observed in goats with patent stomata, urinary bladder marsupialization may result in ascending urinary tract inflammation or infection. CLINICAL RELEVANCE: Based on our results, urinary bladder marsupialization should be recommended with caution as the primary method for management of urinary tract obstruction in clinical cases.

Effects of equine recombinant interleukin-1alpha and interleukin-1beta on proteoglycan metabolism and prostaglandin E2 synthesis in equine articular cartilage explants.

OBJECTIVES: To evaluate the effects of equine recombinant interleukin-1alpha (rEqIL-1alpha) and recombinant interleukin-1beta (rEqIL-1beta) on proteoglycan metabolism and prostaglandin E2 (PGE2) synthesis by equine articular chondrocytes in explant culture. SAMPLE POPULATION: Near full-thickness articular cartilage explants (approx 50 mg) harvested from stifle joints of a 3-year-old and a 5-year-old horse. PROCEDURE: Expression constructs containing cDNA sequences encoding EqIL-1alpha and EqIL-1beta were generated, prokaryotically expressed, and the recombinant protein purified. Near full-thickness articular cartilage explants (approx 50 mg) harvested from stifle joints of a 3-year-old and a 5-year-old horse were separately randomized to receive rEqIL-1alpha or rEqIL-1beta treatments 10 to 500 ng/ml). Proteoglycan release was evaluated by 1,9-dimethylmethylene blue spectrophotometric analysis of explant media glycosaminoglycan (GAG) concentration and release of 35S-sulfate-labeled GAG to explant media. Proteoglycan synthesis was assessed by quantification of 35S-sulfate incorporation into proteoglycan. Explant media PGE2 concentrations were evaluated using a PGE2-specific enzyme-linked immunoassay. Data were collected at 48-hour intervals and normalized by DNA content. RESULTS: Proteoglycan release was induced by rEqIL-1alpha and rEqIL-1beta at concentrations > or =0.1 ng/ml, with 38 to 76% and 88 to 98% of total GAG released by 4 and 6 days, respectively. Inhibition of proteoglycan synthesis (42 to 64%) was observed at IL-1 concentrations > or = 0.1 ng/ml at 2 and 4 days. Increased PGE2 concentrations were observed at IL-1 concentrations > or = 0.1 ng/ml at 2 and 4 days. CONCLUSIONS AND CLINICAL RELEVANCE: The rEqIL-1 induced potent concentration-dependent derangement of equine chondrocyte metabolism in vitro. These findings suggest this model may be suitable for the in vitro study of the pathogenesis and treatment of joint disease in horses.

Evaluation of lameness associated with arthroscopy and arthrotomy of the normal canine cubital joint.

OBJECTIVE: To evaluate gait, range of motion (ROM), and lameness in normal dogs after arthroscopy or arthrotomy of the cubital joint. STUDY DESIGN: Experimental study. ANIMALS: Fourteen mature, healthy dogs. METHODS: Dogs were randomly assigned to treatment groups. Seven underwent arthrotomy and 7 underwent arthroscopy of the left cubital joint. Dogs were evaluated using kinetic gait assessment, subjective evaluation scores, and cubital joint ROM. Evaluations were performed before and on days 2, 4, 7, 15, 22, and 29 after surgery. Radiographs made before and after the surgical procedures were evaluated. RESULTS: Significant differences in peak vertical force and vertical impulse force were not observed between surgery groups (P =.88 and.49, respectively). Joint ROM was not significantly different between groups (P =.09 for flexion and.91 for extension). For all dogs, joint ROM and radiographic evaluations remained normal throughout the study period. Significant differences in subjective lameness scores, weight bearing, or pain were not observed between groups (P >/ or =.19 for all variables). CONCLUSIONS: In this study population, significant differences between dogs undergoing arthroscopy or arthrotomy of the cubital joint with respect to postoperative pain, weight bearing, joint ROM, or temporal improvement of ground reaction forces were not observed. CLINICAL RELEVANCE: The results of this study suggest that postoperative morbidity should not be a factor when making a decision to perform either arthroscopy or arthrotomy for exploration of the medial compartment of the canine cubital joint.