**5. Materials and Methods**

#### *5.1. Animals*

In this study, we used male Wistar rats weighting between 170 and 190 g. The rats were housed in an environment with temperatures of 21 ± 2 ◦C, and light-controlled with a 12 h/12 h light/dark cycle. Standard food and water were available ad libitum. The procedures were performed according to the guidelines for the ethical use of conscious animals in pain research, following the International Association for the Study of Pain [32]. This study was approved by the Institutional Animal Care Committee of the Butantan Institute (CEUAIB, protocol number 9766020419 (date of approval 19 March 2014) and 1245/14 (date of approval 17 April 2019)). The ARRIVE guidelines was followed while conducting the study.
