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We describe a surgical procedure used to induce peripheral ischemia in rabbits with hyperlipidemia and diabetes. This surgery acts as a preclinical model for conditions experienced in peripheral artery disease in patients. Angiography is also described as a means to measure the extent of introduced ischemia and recovery of perfusion.
Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular disease is the development of ischemia. In severe cases, patients can develop critical limb ischemia in which they experience constant pain and an increased risk of limb amputation. Current therapies for peripheral ischemia include bypass surgery or percutaneous interventions such as angioplasty with stenting or atherectomy to restore blood flow. However, these treatments often fail to the continued progression of vascular disease or restenosis or are contraindicated due to the overall poor health of the patient. A promising potential approach to treat peripheral ischemia involves the induction of therapeutic neovascularization to allow the patient to develop collateral vasculature. This newly formed network alleviates peripheral ischemia by restoring perfusion to the affected area. The most frequently employed preclinical model for peripheral ischemia utilizes the creation of hind limb ischemia in healthy rabbits through femoral artery ligation. In the past, however, there has been a strong disconnect between the success of preclinical studies and the failure of clinical trials regarding treatments for peripheral ischemia. Healthy animals typically have robust vascular regeneration in response to surgically induced ischemia, in contrast to the reduced vascularity and regeneration in patients with chronic peripheral ischemia. Here, we describe an optimized animal model for peripheral ischemia in rabbits that includes hyperlipidemia and diabetes. This model has reduced collateral formation and blood pressure recovery in comparison to a model with a higher cholesterol diet. Thus, the model may provide better correlation with human patients with compromised angiogenesis from the common co-morbidities that accompany peripheral vascular disease.
Peripheral arterial disease (PAD) is a common circulatory disorder in which the progression of atherosclerotic plaque formation leads to a narrowing of blood vessels in the limbs of the body. The recent increase in risk factors for atherosclerosis, including diabetes, obesity, and inactivity, has led to increasing prevalence of vascular disease1. Currently, it is estimated that 12%–20% of the general population over 60 years old has peripheral arterial disease2. A major consequence of peripheral arterial disease is the development of peripheral ischemia, most commonly found in the lower limbs. In severe cases, pati....
Studies involving animals were performed with the approval of the University of Texas at Austin and the UTHealth Science Center at Houston Institutional Animal Care and Use Committee (IACUC), the Animal Care and Use Review Office (ACURO) of The United States Army Medical Research and Materiel Command Office of Research Protections, and in accordance with NIH guidelines for animal care.
1. Induction of diabetes and hyperlipidemia
Following induction of diabetes and initiation of the 0.1% cholesterol diet, the total cholesterol for the rabbits with diabetes and cholesterol diet was 123.3 ± 35.1 mg/dL (n = 6 male rabbits) averaged overall time points and rabbits. The BGL level for these rabbits was 248.3 ± 50.4 mg/dL (n = 6 male rabbits). A time course for blood chemistries and leg blood pressure ratios in a typical rabbit is shown in Figure 3 in comparison to rabbits under a higher choles.......
We have presented a preclinical model for inducing hind limb ischemia in rabbits with diabetes and hyperlipidemia. In many studies, there is ambiguity to the technique used to create hind limb ischemia in rabbits. In mice, the severity and recovery from hind limb ischemia is highly dependent on the location the ligation and technique used to induce ischemia. The significance of the technique presented in this work is that it allows for the consistent induction of ischemia that does not fully recover after 8 weeks in diab.......
The authors gratefully acknowledge funding through the Department of Defense Congressionally Directed Research Program (DOD CDMRP; W81XWH-16-1-0582) to ABB and RS. The authors also acknowledge funding through the American Heart Association (17IRG33410888), the DOD CDMRP (W81XWH-16-1-0580) and the National Institutes of Health (1R21EB023551-01; 1R21EB024147-01A1; 1R01HL141761-01) to ABB.
....Name | Company | Catalog Number | Comments |
0.9% Sodium Chloride | Henry Schein Medical | 1537468 / 1531434 | 250 mL bag / 1000 mL irrigation btl |
1 mL Syringe | VWR | BD309628 | |
10 mL Syringe | VWR | BD309695 | |
10% Formalin | Fisher-Scientific | 23-245684 | |
18G Needle | VWR | 89219-294 | |
20G Needle | VWR | 89219-340 | |
25G Needle | VWR | 89219-290 | |
27G Needle | VWR | 89219-288 | |
5 mL Syringe | VWR | BD309646 | |
5% Dextrose | Patterson Veterinary | 07-800-9689 | |
Acepromazine | Patterson Veterinary | VEDC207 | |
Alfaxalone | Patterson Veterinary | 07-891-6051 | |
Alginate | Sigma-Aldrich | PHR1471-1G | |
Alloxan Monohydrate | Sigma-Aldrich | A7413 | |
Angiography Equipment | Toshiba | Infinix-i | |
Angiography Injector | Medrad | ||
Anti-Mouse Ab Alexa 594 | Thermo Fisher Scientific | A-11032 | Secondary Antibody for IHC |
Anti-Rabbit Ab Alexa 488 | Thermo Fisher Scientific | A-11008 | Secondary Antibody for IHC |
a-SMA Antibody | Abcam | ab5694 | Primary Antibody for IHC |
Baytril | Bayer Animal Health | 724089904201 | Enrofloxacin |
Blood Chemistry Panel | IDEXX | 2616 | Rabbit Panel |
Blood Pressure Cuff | WelchAllyn | Flexiport Disposable BP Cuff-infant size 7 | |
Blood Pressure Monitor | Vmed Technology | Vmed Vet-Dop2 | |
Bupivacaine | Henry Schein Medical | 6023287 | |
Buprenorphine | Patterson Veterinary | 42023017905 | |
Buprenorphine SR | ZooPharm | ||
Calcium Sulfate | CB Minerals | Food and Pharmaceutical Grade USP and FCC | |
Chlorhexidine Scrub | Patterson Veterinary | 07-888-4598 | |
Chloroform | Fisher-Scientific | C298-4 | |
Cholesterol | Sigma-Aldrich | C8503 | |
DAPI | Thermo Fisher Scientific | 62248 | |
Ear Vein Catheter | Patterson Veterinary | SR-OX165 | Surflo IV catheters |
Endotracheal tube | Patterson Veterinary | Sheridan Brand, Depends on Rabbit Size | |
Glucometer | Amazon | B001A67WH2 | Accu-Chek Aviva |
Glucometer Test Strips | McKesson Medical-Surgical | 788222 | Accu-Chek Aviva Plus |
Guidewire | Boston Scientific | 39122-01 | |
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 |
Heparin | Patterson Veterinary | ||
Insertion Tool | Merit Medical Systems | MAP550 | metal wire insertion tool |
Insulin | HPB Pharmacy | Novalin R & Novalin N | |
Insulin Syringes | McKesson Medical-Surgical | 942674 | |
Introducer | Cook Medical | G28954 | 3F Check Flo Performer Introducer |
Isoflurane | Henry Schein Medical | 1100734 | |
Ketamine | Patterson Veterinary | 856440301 | |
Lactated Ringers | McKesson Medical-Surgical | 186662 | |
Lidocaine | McKesson Medical-Surgical | 239936 | |
Lidocaine/Prilocaine cream | McKesson Medical-Surgical | 761240 | |
Ligaloop | V. Mueller | CH117 / CH116 | White Mini / Yellow Mini |
Mazola Corn Oil | Amazon | B0049IIVCI | |
Medrad Syringe | McKesson Medical-Surgical | 346920 | 150 mL |
Meloxicam | Patterson Veterinary | ||
Metal ball sutures | Ethicon-Johnson & Johnson | K891H | 4-0 silk C-1 30" |
Metzenbaum Scissors | Fine Science Tools | 14019-13 | |
Midazolam | Henry Schein Medical | 1215470 | |
Nitroglycerin | McKesson Medical-Surgical | 927528 | |
PECAM Antibody | Novus Biologicals | NB600-562 | Primary Antibody for IHC |
Perfusion Pump | Masterflex | ||
Pigtail Catheter | Merit Medical Systems | 1310-21-0053 | 3F pigtail |
Polydioxanone (PDS II) suture | McKesson Medical-Surgical | 129271 | 4-0 taper RB-1 (needle comes on suture) |
Polydioxanone (PDS II) suture | McKesson Medical-Surgical | 129031 | 4-0 reverse cutting FS-2 |
Polyglactin 910 (Vicryl) suture | Butler | 7233-41 | 3-0 taper RB-1 |
Polyglactin 910 (Vicryl) suture | McKesson | 104373 | 4-0 reverse cutting FS-2 |
Rabbit Chow (Alfalfa) | LabDiet | 5321 | |
Rabbit Restrainer | VWR | 10718-000 | |
Rib Cutters | V. Mueller | ||
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" |
Stainless Steel Ball | McMaster-Carr | 1598K23 | 3-mm diameter |
Surgical Drapes | Gepco | 8204S | |
Syringe Pump | DRE Veterinary | Versaflow VF-300 | |
Visipaque contrast media | McKesson Medical-Surgical | 509055 | |
Weitlaner Retractor | Fine Science Tools | 17012-13 |
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