Models of Inflammation: Carrageenan‐ or Complete Freund Adjuvant (CFA)

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Abstract
Table of Contents
Materials
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Literature Cited

Abstract

Animal models of inflammation are used to assess the production of inflammatory mediators at sites of inflammation, the anti?inflammatory properties of agents such as nonsteroidal anti?inflammatory drugs (NSAIDs), and the efficacy of putative analgesic compounds in reversing cutaneous hypersensitivity. This unit details methods to elicit and measure carrageenan? and complete Freund's adjuvant (CFA)?induced cutaneous inflammation. Due to possible differences between the dorsal root sensory system and the trigeminal sensory system, injections of either the footpad or vibrissal pad are described. In this manner, cutaneous inflammation can be assessed in tissue innervated by the lumbar dorsal root ganglion neurons (footpad) and by the trigeminal ganglion neurons (vibrissal pad). Curr. Protoc. Pharmacol. 56:5.4.1?5.4.7. © 2012 by John Wiley & Sons, Inc.

Keywords: inflammation; carrageenan; complete Freund's adjuvant; edema; nociception

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Reagents and Solutions
Commentary
Literature Cited
Figures
Tables

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Materials

Basic Protocol 1:

Materials

Male or female rats weighing >100 g upon arrival
0.5% to 2.0% (w/v) carrageenan solution (see recipe )
Complete Freund's adjuvant (CFA; see recipe )
0.9% (w/v) saline, sterile
70% (v/v) ethanol
Inhalational anesthesia [e.g., 5% (v/v) isoflurane], optional
Betadine

Solid‐bottom cages
Scales, accurate to 0.1 and 0.0001 g
27‐G, ¹ /₂ ‐in needle
1‐ml syringes
Kimwipes
Clear plastic box for the inhalational anesthesia or animal restrainer
6‐mm biopsy punch

Additional reagents and equipment for euthanizing the animal (Donovan and Brown, )

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Figures

Figure 5.4.1 (A ). Site of injection of substances into the footpad of the rat. (B ) Optimal area for paw punch biopsies to obtain consistent results.

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Figure 5.4.2 (A ) Photograph of a footpad from an animal 24 hr after injection with 100 µl saline. (B ) Photograph of a footpad from an animal 24 hr after injection with 100 µl CFA (0.5 mg/ml).

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Figure 5.4.3 Effects of ketorolac on carrageenan‐induced thermal hyperalgesia and edema. (A ) Thermal hypersensitivity in animals injected with saline (100 µl; animal tested 3 hr after injection) or carrageenan (100 µl of 0.5%; animal tested 3 hr after injection). Thermal hypersensitivity is expressed as the paw withdrawal latencies (sec) to a thermal stimulation using the Hargreaves apparatus. Ketorolac (10 mg/kg, i.p.; dark fill) administered 30 min prior to carrageenan injection completely reversed the inflammation‐induced thermal hypersensitivity ( p <0.05 determined by a t ‐test between vehicle‐ and ketorolac‐treated animals, n = 3/group). (B ) Edema induced by saline or carrageenan injection. The edema is expressed as the wet weight (mg) of paw punch biopsy mass. Ketorolac (10 mg/kg, i.p.; dark fill) administered 30 min prior to carrageenan injection partially reversed the inflammation‐induced edema ( p <0.05 determined by a t ‐test between vehicle‐ and ketorolac‐treated animals, n = 3/group).

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Literature Cited

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