An Experimental Model of Peripheral Ischemia: A Procedure to Generate Peripheral Ischemia via Femoral Artery Ligation in a Rabbit

Protocolo

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Isolation of the femoral artery

1. Anesthetize a New Zealand rabbit (4–6 months old) with pre-induced diabetes and hyperlipidemia, using 20–40 mg/kg ketamine and 2 mg/kg midazolam via subcutaneous injection. Place the rabbit on 1.5%–3% isoflurane (typically 2%) throughout the initial sedation using a mask.
2. Make a longitudinal incision in the skin over the right femoral artery using a scalpel (#15 blade). Ensure that the incision extends inferiorly from the inguinal ligament ending at the area just proximal to the patella (approximately 6 cm).
3. Use blunt dissection with curved Metzenbaum scissors or a curved mosquito hemostat to expose the femoral artery.
4. Use Weitlaner retractors to hold the incision open.
5. Add 0.5 mL of 1% lidocaine locally to reduce nerve irritation and promote vasodilation.
6. Continue blunt dissection of the tissues to free the entire length of the femoral artery along with all branches of the femoral artery, including the inferior epigastric, deep femoral, lateral circumflex, and superficial epigastric arteries (Figure 1A).
7. Dissect further along the popliteal and saphenous arteries as well as the external iliac artery (Figure 1A). Periodically moisten the area with saline to protect from tissue damage. If the blunt dissection is performed along the femoral groove (between the muscles) there is no need to cut the muscle.
8. Carefully separate the artery from the vein and nerve as shown in Figure 1B,C. Ligate the arteries indicated by the diagram with 4.0 silk sutures by placing two ties with enough space between them to cut the artery. These ties are performed with a Surgeon's knot followed by four square knots.
9. Cut between the two ties on the ligated arteries using the small Metzenbaum scissors. Excise the femoral artery from its proximal origin as a branch of the external iliac artery to the point distally where it bifurcates to form the saphenous and popliteal arteries.

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Resultados

Figure 1: Induction of hind limb ischemia in rabbits through femoral artery ligation and excision. (A) Illustration of the vascular anatomy of the rabbit hind limb. Place ties at all the points marked to ligate the arteries. (B) Surgical field showing the cut down to the femoral artery prior to ligation. (C) Femoral arteries with ligations in place to induce hind limb ischemia.

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Materiais

Name	Company	Catalog Number	Comments
0.9% Sodium Chloride	Henry Schein Medical	1537468 / 1531434	250 mL bag / 1000 mL irrigation btl
10 mL Syringe	VWR	BD309695
27G Needle	VWR	89219-288
Acepromazine	Patterson Veterinary	VEDC207
Alfaxalone	Patterson Veterinary	07-891-6051
Buprenorphine SR	ZooPharm
Hair Clippers	Amazon	B000CQZI3Q	Oster #40 blade
Heating Pad	Cincinnati Subzero	273
Heating Pad Pump	Gaymar		Gaymar T/Pump
Hemostat	Fine Science Tools	13009-12	Curved Mosquito Hemostat
Isoflurane	Henry Schein Medical	1100734
Ketamine	Patterson Veterinary	856440301
Lidocaine	McKesson Medical-Surgical	239936
Lidocaine/Prilocaine cream	McKesson Medical-Surgical	761240
Metzenbaum Scissors	Fine Science Tools	14019-13
Midazolam	Henry Schein Medical	1215470
Scalpel	Fine Science Tools	10003-12
Scalpel Blade	Fine Science Tools	10015-00	#15 blade
Silk Sutures	Ethicon-Johnson & Johnson	A183H	4-0 silk ties 18"
Surgical Drapes	Gepco	8204S
Weitlaner Retractor	Fine Science Tools	17012-13