Systat 12

In order to allow a direct comparison, much of the methodology of these studies is identical to our previous report which used bladder re-inflammation as an adult insult^{29 (link)} rather than hind limb inflammation. Anesthetics, drug doses, timing and other variables were therefore chosen to be consistent with that other report. Studies were performed in 171 female Sprague-Dawley rats and were approved by the University of Alabama at Birmingham’s Institutional Animal Care and Use Committee and followed the ethical guidelines of the International Association for the Study of Pain^{53 (link)}. To obtain female rat pups, timed pregnant females were obtained from Envigo Laboratories (Formerly Harland, Indianapolis, IN) and date of birth verified by daily observation of cages. As pups, separate groups of the female pups underwent treatments for three consecutive days on P14–16. Following each treatment, pups were returned to home cages. Subsequently, rats were raised using standard husbandry methods with weaning from the dams at three to four weeks of age. All rats were raised to 12–15 weeks of age and then underwent additional testing. For statistical analyses, the SYSTAT 12 (SPSS, Inc. San Jose, CA, United States) software package was utilized.

Descriptive statistics are reported as group mean ± SEM. Statistical comparisons were made using one way and/or repeated measures analysis of variance (ANOVA) with Tukey’s HSD used for pairwise post hoc comparisons (Systat 12; SPSS, Inc. San Jose, CA, USA). Statistical significance for all analyses was set at p≤0.05.