Assessment of Allodynia Relief by Tissue-Protective Molecules in a Rat Model of Nerve Injury-Induced Neuropathic Pain - Tissue-Protective Cytokines: Methods and Protocols - page 185

17. Student iris scissors, straight 11.5 cm (Fine Science Tools,
Heidelberg, Germany).
18. Halsey needle holder, smooth (Fine Science Tools, Heidelberg,
Germany).

1. Plateau with grid (UGO Basile, Varese, Italy).
2. Perspex cages with lid (UGO Basile, Varese, Italy).
3. Semmes-Weinstein monofilaments (North Coast Medical
Inc., San Jose, CA, USA).

2.2 Assessment
of Pain: Tactile
Allodynia

1. Syringe (1 ml).
2. Needle 25 G, bent 90°.
3. Acetone, analytical grade.

2.3 Assessment
of Pain: Cold Allodynia

1. PBS.
2. ARA290 (Araim Pharmaceuticals, Ossining, NY, USA).
3. Syringe, 1 ml equipped with 25 G needle.

2.4 Treatment
with ARA290

3 Methods

3.1 Induction
of the Neuropathic
Pain Model: Spared
Nerve Injury

1. Sterilize the instruments, for instance with a tabletop
sterilizer.
2. Disinfect the surgical area of the table with 70% ethanol.
3. Place an absorbing under pad on the surgical area and place
the surgical tools.
4. Fifteen minutes prior to surgery, administer a single dose of
0.01-0.05 mg/kg buprenorphin subcutaneously in the scruff
of the neck for the relief of acute post operative pain.
5. To start surgery, induce and maintain anesthesia (6% induc-
tion, 3% maintenance in medicinal air mixture).
6. Place animal on the stomach and shave the leg that is going to
be operated on.
7. Disinfect the shaved hind leg and direct and fixate it with a
piece of tape towards yourself.
8. Draw an imaginary line between the patella and the crest of
the ilium and locate the center of the line. This is approxi-
mately where the trifurcation of the nerve is situated
(Fig. 1a ).
9. Lift the skin of the hind leg with the standard pattern
forceps.
10. Make a small incision with the Metzenbaum scissors perpen-
dicular to the imaginary line 1 cm distally from where the
trifurcation is supposed to be.