Formalin Model Research Articles

Morphine, clonidine coadministration in subanalgesic doses: effective control of tonic pain.

The aim of the present study was to investigate whether clonidine and morphine interact positively to produce analgesia against the low intensity tonic pain represented by the formalin model in rats. Sub-threshold doses of morphine (0.5 mg kg-1) and clonidine (0.025 mg kg-1) were found to elicit marked antinociceptive effects when co-administered intraperitoneally, 15 min prior to formalin challenge. Repeated administration of this combination for eight days did not exhibit any significant decay of this analgesic response, whilst morphine (2 mg kg-1)-induced analgesia, deteriorated after similar administration. Clonidine and morphine thus exhibit a supra-additive effect against low intensity pain with negligible potential for induction of tolerance. This finding may be relevant for the long term control of chronic pain in certain clinical conditions.

Nitrous oxide induces preemptive analgesia in the rat that is antagonized by halothane.

Noxious stimulation-induced sensitization of the central nervous system has been proposed as a key element in the development of subsequent protracted pain. Accordingly, the authors used the formalin model of pain to test the hypothesis that general anesthesia can produce preemptive analgesia and thereby interfere with noxious stimulation-induced central sensitization. Rats received 0.9% or 1.8% halothane, 30% or 75% nitrous oxide (N2O), or 75% N2O plus 0.9% halothane (n = 4 or 5 per group). Control rats (n = 5) received only 100% oxygen. Fifteen minutes after the induction of anesthesia, formalin was injected subcutaneously into a hind paw of each rat, and anesthesia was maintained for 5 more min. Because the behavioral pain response to formalin (i.e., flinching of the injected paw) is biphasic, these treatment groups were anesthetized only during phase 1 (acute phase). Another group (n = 5) received 75% N2O only during phase 2 (delayed phase). Reversibility of the N2O effect was tested by the administration of naloxone before phase 1 or naltrexone during phase 2 (n = 5 per group). Finally, additional rats anesthetized as described above (n = 4 or 5 per group) underwent tail-flick testing during anesthesia. All anesthetics reduced phase 1 pain behavior, but only N2O produced antinociception on tail-flick testing. Thirty percent and 75% N2O, administered during phase 1, suppressed phase 2 flinching 29% and 49%, respectively, whereas nitrous oxide administered after phase 1 did not suppress phase 2 pain behavior. This effect of nitrous oxide was reversed by an opioid antagonist given during phase 1 but not phase 2. Halothane administered during phase 1 had no effect on phase 2 flinching, and it antagonized the effect of 75% N2O. Nitrous oxide induces dose-dependent preemptive analgesia in this model that is reversed partially by naloxone, thus suggesting the involvement of endogenous opioids in this action. In contrast, halothane has no preemptive analgesic properties and even antagonizes the analgesic effect of nitrous oxide. Hence, the hypnotic potency of an anesthetic is a poor indication of its preemptive analgesic potential.

Synergistic anti-nociceptive effect of L-NG-nitro arginine methyl ester (L-NAME) and flurbiprofen in the mouse.

1. L-NG-nitro arginine methyl ester (L-NAME) administered i.p. produces anti-nociception in the mouse assessed by the formalin-induced paw licking and acetic acid-induced abdominal constriction models. The non-steroidal anti-inflammatory drug (NSAID), flurbiprofen, was similarly anti-nociceptive in both models. 2. Combination of a sub-threshold dose of L-NAME (10 mg kg-1) with increasing doses of flurbiprofen (25- 75 mg kg-1) or a sub-threshold dose of flurbiprofen (50 mg kg-1) with increasing doses of L-NAME (10- 100 mg kg-1) resulted in potentiated anti-nociception in the formalin model. Combined therapy with sub-threshold doses of L-NAME (10 mg kg-1) and indomethacin (10 mg kg-1) also resulted in significant anti-nociception. In addition, combining sub-threshold doses of L-NAME (12.5 mg kg-1) and flurbiprofen (2 mg kg-1) significantly reduced acetic acid-induced abdominal constriction. 3. L-NAME (10 mg kg-1) administered i.p. caused a significant (approximately 35%) increase in MAP in the urethane-anaesthetized mouse. Flurbiprofen (50 mg kg-1) was inactive. Combination treatment with L-NAME (10 mg kg-1) and flurbiprofen (50 mg kg-1) failed to elevate MAP above that observed with L-NAME alone. Neither L-NAME (10 mg kg-1) nor flurbiprofen (50 mg kg-1) either alone or in combination significantly altered mouse locomotor activity. 4. These results suggest that L-NAME and flurbiprofen/indomethacin act synergistically in their anti-nociceptive action in the mouse. Combination therapy with L-NAME and flurbiprofen and a similar NSAID may provide an alternative to the clinical control of pain in man.

Neurogenic component of different models of acute inflammation in the rat knee joint.

This study was performed to investigate whether different models of acute joint inflammation showed a neurogenic component and to establish whether this is mediated through sensory afferent or sympathetic efferent nerve fibres. Intra-articular injection of 2% carrageenan, 20 micrograms substance P, 1% formalin, and 2% urate all produced an inflammatory response. Prior surgical denervation of the joint significantly inhibited this response in the carrageenan and formalin models, but not the others. Pretreatment of the joint with 1% capsaicin (about one week previously) significantly reduced the inflammatory response in all models except formalin. In animals pretreated long term with reserpine (to deplete sympathetic nerve endings of their neurotransmitters) significant reductions occurred in the inflammatory responses to substance P and urate. Intraarticular injection of compound 48/80 produced a marked inflammatory response, which was only significantly reduced by capsaicin pretreatment. These results suggest that both the formalin and carrageenan models of inflammation depend to some extent on the integrity of the sensory innervation of the joint, and thus have a neurogenically mediated component to the inflammatory process they generate. In these models there seems to be little contribution from sympathetic efferent fibres. Each model of inflammation showed a different pattern of response to the pretreatments, suggesting that the mediators of the inflammatory process may differ in each case.

Neurokinin and NMDA antagonists (but not a kainic acid antagonist) are antinociceptive in the mouse formalin model

While much evidence implicates substance P (SP), an endogenous neurokinin (NK), as a primary sensory transmitter of acute pain in mammalian spinal cord, its role in continuous (tonic) pain is less clear. Although glutamate is co-localized with SP in dorsal root ganglion neurons, its role in nociceptive processing is uncertain. While antagonists of NKs and excitatory amino acids (EAAs) have been found to be antinociceptive in some acute assays, they have not been tested against tonic pain. We hypothesize that: 1. (1) NKs and EAAs contribute to signaling of tonic chemogenic nociception 2. (2) interaction between NK and EAA systems is important in determining the perceived intensity of a continuous noxious stimulus. We therefore evaluated two NK antagonists ([ d-Pro 2, d-Trp 7,9] SP (DPDT-SP, 0.26–6.6 nmoles, non-specific) and [ d-Pro 4, d-Trp 7,9,10,Phe 11]-SP(4–11) (DPDTP-octa, 1.6–12.3 nmoles, somewhat NK-1 selective)), as well as dl-2-amino-5-phosphonovalerate ( dl-AP5, NMDA antagonist, 0.05–1 nmole) and urethane (a kainic acid (KA) antagonist at 2.5 μmoles) for antinociceptive activity in the mouse formalin model. Administered intrathecally (i.t.), dl-AP5 and both NK antagonists were significantly antinociceptive while urethane (2.5 μmoles) and naloxone (2.7 nmoles) were inactive. A 50 values for mean % analgesia, nmoles/mouse i.t. (95% CLs) were: DPDT-SP, 1.1 (0.79–1.6); DPDTP-octa, 3.9 (2.4–6.1); dl-AP5, 0.29 (0.16–0.71). The antinociception associated with 1.3 nmoles of DPDT-SP was not reversed by co-administering 2.7 nmoles of naloxone. Co-administration of 0.1 nmoles of dl-AP5 with either 1.3 nmoles of DPDT-SP or 3.3 nmoles of DPDTP-octa did not lead to additive antinociception. One nmole of dl-AP5 also produced significant antinociception in the mouse tail-flick test. These data support the contention that NK (probably NK-1) and NMDA (but not KA) receptors are involved in spinal processing of tonic chemical/inflammatory nociception in mice.

Methodological refinements to the mouse paw formalin test: An animal model of tonic pain

The formalin tonic pain test has recently been adapted for use in mice. We have observed two problems with this procedure. First, compared to a vehicle control group, the majority of mice injected in a hind paw with dilute formalin exhibited a significant nociceptive response (paw licking), including peaks of activity, well beyond the end of the recommended observation period; and, second, an unacceptably wide variation in response occurred. We report two modifications that eliminate these difficulties: a) extending the observation period to 20–45 min postformalin injection, and b) lightly anesthetizing the mice with ether before injection of formalin into the paw. The modified protocol was tested by evaluating morphine, pentazocine and zomepirac (analgesic A 50 values were 2.1, 23.8, and > 100 mg/kg, s.c., respectively). An intrathecal or intracerebroventricular injection of saline, given to the lightly anesthetized mouse, did not interfere with the nociceptive behavior; this finding widens the applicability and usefulness of the test. Collectively, these refinements significantly improve the mouse formalin model of tonic pain.