Hot-plate analgesia testing

Capsaicin- and vehicle-treated mice were subjected to hot-plate analgesia testing at 4, 8, and 12 weeks of age (before the start of tibial compression) to determine response time to a constant thermal stimulus of 55°C as previously described [36]. Mice were placed on a hot-plate (LE 7406, Coulborn Instruments, Whitehall, PA) and removed after indication of discomfort, determined as twitching or licking of a hind limb or jumping, or after a maximum of 30 seconds, and the latency time of the response was recorded. Each mouse was tested twice at each age, with approximately 2 hours between tests, and the latency times were averaged for each mouse.