Dose-Finding in the Development of an LPS-Induced Model of Synovitis in Sheep

Models of transient synovitis that can be controlled with antiinflammatory and analgesic drugs have been used to study pain amelioration. To this end, we aimed to determine the dose of intraarticularly administered E. coli LPS that induced signs of synovitis without systemic signs in clinically healthy male castrated sheep (n = 14). In phase 1, a single dose of LPS (0.5, 1.0, 1.5, or 2.0 ng in a total volume of 0.5 mL) was administered into the right stifle joint. In phase 2, a dose of LPS (1.0 or 2.0 μg) in 0.3 mL was administered to 4 naïve sheep. In phase 3, 4 sheep from phase 1 were inoculated after a 60 d washout period with either 0.5 or 1.0 μg of LPS. During the first 48 h after LPS administration, the following were performed: assessment of clinical parameters; scoring for lameness, pain on limb flexion, and local swelling; and ultrasonography of the joints were performed. The doses tested during phase 1 produced subtle signs. During phase 2, mild to moderate lameness with no evidence of systemic signs occurred at both doses. In phase 3, clinical responses were similar between the 0.5- and 1-µg doses. Signs of swelling were not observed at any time. Therefore, we consider the 0.5-µg to be the most appropriate for this model, because it was the lowest dose tested capable of causing lameness without signs of systemic inflammation in all animals.

As an experimental model for the study of arthropathies, the aseptic administration of small doses of endotoxin in the joint induces mild to moderate inflammation and the development of clinical signs similar to those of the naturally occurring disease.⁶ Some studies have used models of transient synovitis to determine whether the associated pain can be controlled with antiinflammatory and analgesic drugs. The use of an LPS-induced model of synovitis to evaluate the analgesic effect of various therapeutic protocols has mainly been reported for horses.^9,16,27,28 However, sheep are an important model species in biomedical research, particularly in orthopedic studies,^15,20,32 due to their similarity in weight, size, and joint and bone structure with humans, and in cardiovascular^7,11 studies, because they are good models of cardiac anatomy and physiology. Consequently, the development of analgesia protocols for acute pain conditions is greatly needed.Animal experiments are under increasing focus regarding their ethical and legal aspects. In vivo studies are permitted when methods consistent with the 3Rs principals (replacement, refinement, and reduction) are considered and implemented.²⁶ This means that experiments have to be performed without animals when possible (replacement) or with as few animals as possible (reduction) and with as little pain and distress as possible (refinement). In this context, species-specific analgesia is considered an important refinement method applicable to the majority of research.²⁵ However, few studies have been conducted to determine analgesia protocols for different pain conditions in sheep. The standardization of animal pain models is necessary for the reliable evaluation of efficient and different drug protocols.^10,19,24To guide standardization of the dose of E. coli LPS for intraarticular administration, with the aim of developing a pain model for studies of analgesia in sheep, we here assessed the ability of various intraarticular doses of LPS to trigger synovitis. Our hypothesis was that the dose established for use in horses (0.5 ng/joint) would trigger similar effects in sheep.