Analgesic Activity of Tramadol and Buprenorphine after Voluntary Ingestion by Rats (Rattus norvegicus).

Effective pain management for rats and mice is crucial due to the continuing increase in the use of these species in biomedical research. Here we used a recently validated operant orofacial pain assay to determine dose-response curves for buprenorphine and tramadol when mixed in nut paste and administered to male and female rats. Statistically significant analgesic doses of tramadol in nut paste included doses of 20, 30, and 40 mg/kg for female rats but only 40 mg/kg for male rats. For male rats receiving buprenorphine mixed in nut paste, a significant analgesic response was observed at 0.5 and 0.6 mg/kg. None of the doses tested produced a significant analgesic response in female rats. Our results indicate that at the doses tested, tramadol and buprenorphine produced an analgesic response in male rats. In female rats, tramadol shows a higher analgesic effect than buprenorphine. The analgesic effects observed 60 min after administration of the statistically significant oral doses of both drugs were similar to the analgesic effects of 0.03 mg/kg subcutaneous buprenorphine 30 min after administration. The method of voluntary ingestion could be effective, is easy to use, and would minimize stress to the rats during the immediate postoperative period.

Assessment of an Orofacial Operant Pain Assay as a Preclinical Tool for Evaluating Analgesic Efficacy in Rodents.

A model system capable of providing clinically relevant analgesic doses with minimal trauma has been elusive in laboratory animal medicine. Our laboratory has developed an orofacial operant pain system that effectively discriminates between non-noxious and noxious thermal stimuli in rats and mice. Male and female rats (Crl:SD) and mice (Crl:SKR-HR(hr)) were trained to perform a task (placing their face through an opening and having their cheeks stay in contact with thermodes) to receive a reward (a solution of sweetened condensed milk). Currently accepted doses of buprenorphine were tested by using a crossover design. Pain was induced in both species by sensitizing the depilated skin over both cheeks with capsaicin cream or by creating a surgical incision (rats only) and then allowing the animals to contact a temperature-regulated thermode while obtaining a reward. Optimal antinociceptive doses included 0.05 and 0.1 mg/kg in male mice but only 0.05 mg/kg in female mice. In rats, optimal antinociceptive doses included 0.03 and 0.05 mg/kg for male rats but only 0.03 mg/kg for female rats. The 2 pain-induction models in rats (capsaicin cream and surgical incision) did not differ. Our orofacial operant pain assay can determine clinically relevant analgesic doses for rodents in a preclinical assay. The automated, investigator-independent nature of the assay, in conjunction with its high sensitivity, makes this method an improvement over traditional noninvasive methods, providing better data for developing optimal analgesic recommendations for rats and mice.

A comparison of two methods of endoscopic dilation of acute subglottic stenosis using a ferret model.

OBJECTIVES/HYPOTHESIS: Balloon dilation is accepted as a first line treatment of acute subglottic stenosis, but its effects on the subglottic tissue remain largely unknown. We aimed to develop an animal model of acute subglottic stenosis using endoscopic techniques. Once developed, this model was used to compare the immediate effects of balloon dilation and endotracheal tube dilation on subglottic tissue. STUDY DESIGN: Prospective randomized animal study. METHODS: Acute subglottic injury was induced in 10 ferrets by endoscopic cauterization with silver nitrate. After 48-72 hours of observation, eight animals were randomized to undergo subglottic dilation with either a 5-mm balloon or endotracheal tubes of increasing diameter. These eight ferrets were euthanized within 10 minutes after dilation. The other two ferrets served as controls and were euthanized following observation only. The larynx from each ferret was harvested, and the subglottis was examined histologically by a pathologist blinded to the treatment arms. RESULTS: Acute subglottic stenosis was induced in all 10 ferrets using the endoscopic technique. Both balloon and endotracheal tube dilation resulted in comparable improvement in the subglottic airway diameter. A decreased thickness of submucosa/lamina propria was seen in the balloon dilation group. CONCLUSIONS: Acute subglottic stenosis can be reliably induced in ferrets using endoscopic techniques. Multiple dilation methods can be used to relieve acute obstruction. Balloon dilators seem to improve airway patency, in part, by decreasing the thickness of the submucosa and lamina propria. Further research is needed to determine how this impacts later stages of wound healing and final outcomes.

Effect of a short-term fast on ketamine-xylazine anesthesia in rats.

Although ketamine-xylazine (KX) anesthesia is commonly used in rats, it is often reported to have an inconsistent anesthetic effect, with a prolonged induction time, an inadequate anesthetic plane, or a very short sleep time. Blood flow to the liver is known to shift after a meal in rats, perhaps explaining anesthetic variability among rats with variable prandial status. The current study tested the hypothesis that a short period of fasting (3 h) prior to induction with intraperitoneal KX anesthesia would provide a shorter time to recumbency, a longer total sleep time, and a more consistent loss of toe pinch response than would fed rats. Two groups of male Sprague-Dawley rats were used in blinded, crossover experiments. KX anesthesia was administered at 2 different doses (50 mg/kg-5 mg/kg and 70 mg/kg-7 mg/kg) after ad libitum feeding or a 3-h fast. There were no significant differences between groups in induction time, total sleep time, or loss of toe pinch response. We conclude that fasting rats for 3 h prior to KX intraperitoneal anesthesia does not affect induction time, total sleep time, loss of toe pinch response or reduce KX anesthetic variability in male Sprague-Dawley rats.

Adeno-associated virus-mediated correction of a canine model of glycogen storage disease type Ia.

Glycogen storage disease type Ia (GSDIa; von Gierke disease; MIM 232200) is caused by a deficiency in glucose-6-phosphatase-alpha. Patients with GSDIa are unable to maintain glucose homeostasis and suffer from severe hypoglycemia, hepatomegaly, hyperlipidemia, hyperuricemia, and lactic acidosis. The canine model of GSDIa is naturally occurring and recapitulates almost all aspects of the human form of disease. We investigated the potential of recombinant adeno-associated virus (rAAV) vector-based therapy to treat the canine model of GSDIa. After delivery of a therapeutic rAAV2/8 vector to a 1-day-old GSDIa dog, improvement was noted as early as 2 weeks posttreatment. Correction was transient, however, and by 2 months posttreatment the rAAV2/8-treated dog could no longer sustain normal blood glucose levels after 1 hr of fasting. The same animal was then dosed with a therapeutic rAAV2/1 vector delivered via the portal vein. Two months after rAAV2/1 dosing, both blood glucose and lactate levels were normal at 4 hr postfasting. With more prolonged fasting, the dog still maintained near-normal glucose concentrations, but lactate levels were elevated by 9 hr, indicating that partial correction was achieved. Dietary glucose supplementation was discontinued starting 1 month after rAAV2/1 delivery and the dog continues to thrive with minimal laboratory abnormalities at 23 months of age (18 months after rAAV2/1 treatment). These results demonstrate that delivery of rAAV vectors can mediate significant correction of the GSDIa phenotype and that gene transfer may be a promising alternative therapy for this disease and other genetic diseases of the liver.

Effects of an enrichment device on voluntary alcohol consumption on single-housed rats.

We evaluated the effect of an enrichment device (that is, a polyurethane bone) on the voluntary consumption of ethanol containing gel by single-housed rats. Male Sprague-Dawley rats (n = 5 per group) were exposed for 4 d to each of the following 3 treatments: access to a new synthetic bone and ethanol gel for 1 h daily (treatment 1); a new bone was left in the cage for 24 h, with access to ethanol gel for 1 h daily (treatment 2); and both the bone and ethanol gel remained in the cage for 24 h (treatment 3). Average alcohol consumption over 4 d was 0.86 +/- 0.13, 0.99 +/- 0.13, and 5.19 +/- 0.37 g/kg in the absence of the bone for treatments 1, 2, and 3, respectively, and 1.00 +/- 0.13, 0.620 +/- 0.07, and 5.55 +/- 0.38 g/kg with the bone for treatments 1, 2 and 3, respectively; none of these values differed significantly with regard to presence of the bone. During treatment 1, time spent with the synthetic bone was highest on the first 2 d, which altered the rate of ethanol consumption but not the total amount of ethanol consumed. During treatments 2 and 3, the rate and amount of ethanol consumption were comparable to basal levels. We conclude that adding an enrichment device that rats can chew and manipulate does not alter ethanol gel consumption. If used, environmental enrichment techniques should be evaluated during the research planning stages to avoid unintended alterations in the response to variables of interest.