OBJECTIVE To evaluate the effect of lipopolysaccharide (LPS) on apoptosis of equine neutrophils in vitro.
SAMPLE Venous blood samples from 40 adult horses.
PROCEDURES Neutrophils were isolated from blood samples and cultured with or without LPS from Escherichia coli O55:B5 for 12 or 24 hours. Neutrophil apoptosis was assessed by use of cytologic examination, annexin V and propidium iodide staining quantified with flow cytometry, coincubation with inducers of intrinsic and extrinsic apoptosis or a toll-like receptor (TLR) 4 inhibitor, and measurement of caspase-3, -8, and -9 activities.
RESULTS Treatment with LPS resulted in a significant delay in apoptosis after incubation for 12 and 24 hours (neutrophils from blood samples of 40 horses). There was a significant correlation between increases in LPS dose and decreases in apoptosis after incubation for 24 hours (3 experiments, each of which involved neutrophils obtained from the same 3 horses at 3 separate times). Caspase-9 activity, but not caspase-3 or -8 activity, was significantly reduced in LPS-treated neutrophils after incubation for 12 hours (neutrophils from blood samples of 17 horses). Treatment with a TLR4 inhibitor or intrinsic and extrinsic inducers of apoptosis prevented LPS-delayed apoptosis.
CONCLUSIONS AND CLINICAL RELEVANCE LPS treatment delayed apoptosis of equine neutrophils in vitro for up to 24 hours in a dose-dependent manner by alteration of the intrinsic pathway of apoptosis and was dependent on TLR4 signaling. Increased neutrophil life span may contribute to the development of a systemic inflammatory response syndrome in endotoxemic horses.
OBJECTIVE To determine the effect of arthrotomy alone or in combination with osteotomy of the proximal portion of the tibia on blood delivery to the patellar tendon of dogs.
SAMPLE 24 canine cadavers.
PROCEDURES One hind limb from each cadaver was assigned to 1 of 4 treatment groups: medial arthrotomy (MA; MA group), lateral arthrotomy (LA; LA group), MA and LA with tibial tuberosity transposition (MALA group), and MA with tibial plateau leveling osteotomy (TPLO; TPLO group). The contralateral hind limb served as the control sample. Contrast solution (barium [33%], India ink [17%], and saline [0.9% NaCl] solution [50%]) was injected through an 8F catheter inserted in the caudal portion of the abdominal aorta. Limbs were radiographed to allow examination of vascular filling. The patella, patellar tendon, and tibial crest were harvested, radiographed to allow examination of tissue vascular filling, and fixed in 4% paraformaldehyde. Vessels perfused with contrast solution were counted in sections obtained from the proximal, middle, and distal regions of each patellar tendon.
RESULTS Vessel counts did not differ significantly among the 3 tendon regions. Compared with results for the control group, delivery of contrast solution to the patellar tendon was significantly decreased in the MALA and TPLO groups but was not changed in the MA or LA groups.
CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that surgical procedures used to treat cranial cruciate injuries (ie, TPLO) and patellar luxation decreased blood delivery to the patellar tendon of canine cadavers, at least acutely.
Objective—To investigate the relationship between inflammatory responses of the temporomandibular joint (TMJ) and the metacarpophalangeal (MCP) joint in clinically normal horses.
Animals—7 mature horses.
Procedures—In each horse, 1 TMJ and 1 MCP joint were injected with lipopolysaccharide (LPS; 0.0025 μg). The contralateral TMJ and MCP joint were injected with saline (0.9% NaCl) solution. Synovial fluid samples were collected from all 4 joints over 24 hours after injection. Concentrations of interleukin-6, tumor necrosis factor-α, transforming growth factor-β, and total protein were measured via immunoassay. Horses were assessed for clinical signs of joint inflammation at each time point.
Results—Concentrations of interleukin-6 were not significantly different between LPS-injected MCP joints and TMJs at any time point. Transforming growth factor-β concentrations were significantly increased in MCP joints, compared with concentrations in TMJs, at 12 and 24 hours after injection. Tumor necrosis factor-α concentrations were significantly higher in LPS-injected TMJs than in LPS-injected MCP joints at 1 and 6 hours after injection. Total protein concentration did not differ significantly between LPS-injected MCP joints and TMJs. Injection of LPS induced clinical inflammation at all time points; additionally, 2 MCP joints (but no TMJs) had an inflammatory response to injection of saline solution.
Conclusions and Clinical Relevance—The inflammatory response to LPS appeared to be attenuated more quickly in TMJs than in MCP joints of horses. The difference in response suggested that a lack of clinical osteoarthritis in the TMJ of horses could be attributable to a difference in cytokine response.