ARA290 in a Rat Model of In fl ammatory Pain - Tissue-Protective Cytokines: Methods and Protocols - page 210

2.4 Assessment
of Pain: Mechanical
Allodynia

1. Raised platform with wire mesh floor and Perspex animal
enclosures with lids.
2. von Frey filaments (catalogue number: 37450; Ugo Basile,
VA, Italy).

2.5 Treatment with
ARA290

1. Saline, 0.9%.
2. ARA290 (Araim Pharmacueticals, Ossining, NY, USA).
3. Syringe, 1 ml with 25 gauge needle.
4. Eppendorf tube (Sterile), 1.5 ml.
5. Sterile filter, 0.22 μ m (Millipore, MA, USA), and syringe,

1 ml.

3 Methods

3.1 Preparation
of Complete Freund's
Adjuvant

1. Sterile filter 0.5 ml saline, using a 0.22 m Millipore filter

attached to a 1 ml syringe, into a 1.5 ml sterile Eppendorf
tube.
2. Vortex the complete Freund's adjuvant and add 0.5 ml of the
adjuvant to the saline using a 1 ml syringe.
3. Vortex the final solution.

μ

1. A sterile gown should be worn for all surgical procedures.
2. All instruments, gauze, and cotton-tipped applicators should
be autoclaved prior to use.
3. Surgical area should be cleaned with 70% alcohol or
disinfectant.
4. Place instruments onto a sterile paper sheet, such as the inside
of an autoclave bag.
5. Using a size 10 scalpel blade, cut a small strip of Gelfoam
(10 × 3 × 3 mm) in preparation for wrapping around the
nerve.
6. Switch heated blanket onto low setting 15 min prior to surgery
and place a sterile sheet onto the blanket.
7. Induce and maintain anesthesia in induction chamber
(isoflurane in oxygen; 3% induction (4 l/min O 2 ) and 1.75%
(0.8 l/min O 2 ) maintenance).
8. Once the rat is anesthetized, place prone onto the heated blan-
ket with the nose into the nose cone.
9. Prior to commencing surgery, check depth of anesthesia (see
Note 2).
10. Place 2-3 gauze swabs under the hind limb/thigh to be oper-
ated on. This will raise the limb and improve visibility for
surgery.

3.2 Induction
of the Neuritis Model
( see Note 1 )