4.8. Behavioural Experiments

The old allodynia method was used to evaluate the tail flick latency. The animal’s tail’s dorsum was kept in a cup of 10 °C cold water and time taken to flick its tail to avoid the painful stimulation was recorded. The cutoff time of 15 s was to be considered to prevent any injury. This was performed on the 0th, 4th, and 8th week of the study before sacrificing the animal [55].

A hot plate at 55 °C was used to measure the hyperalgesic response to thermal nociceptive stimuli on the 0th, 4th, and, 8th week of the study. The time taken by an animal to lick its paws for the first time or jump to protect itself from heat was used to assess the pain threshold [56].