The orofacial formalin test

The subcutaneous injection of formalin into the rat upper lip generates behavioral responses that last several minutes. The time course of the response is similar to what is observed following formalin injection into the paw, i.e. biphasic, with an early and short-lasting first phase followed, after a quiescent period by a second, prolonged (tonic) phase. The applied chemical stimulus (formalin) can be qualified as noxious since it produces tissue injury, activates Aδ and C nociceptors as well as trigeminal and spinal nociceptive neurons and is felt as painful in man. In addition, increasing the concentration of formalin causes a parallel aggravation of histological signs of tissue inflammation and injury. The measured behavioral response (face rubbing) is a relevant end-point: prolonged face rubbing is evoked by formalin but not saline injection and a positive relationship between the amplitude of the response and the formalin concentration is observed, at least up to 2.5%. At higher formalin concentrations, the use of other or additional end-points should be considered. Finally, the behavioral response in the orofacial formalin test is sensitive to various opioid and non-opioid analgesics. The orofacial formalin test can then be considered as a reliable way of producing and quantifying nociception in the trigeminal region of the rat.