Method Article

Reliable Creation of a Murine Abdominal Aortic Aneurysm Model Using Elastin Sensitization and Perivascular Elastase

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DOI:

10.3791/72015

June 26th, 2026

In This Article

Summary

This protocol provides a repeatable and reliable method for inducing abdominal aortic aneurysms (AAAs) in C57BL/6 mice using elastin sensitization and perivascular elastase application, as an AAA model that incorporates an immune-sensitization component.

Abstract

Degradation of the elastic lamellar matrix in the abdominal aortic wall is a histopathologic feature of abdominal aortic aneurysms (AAA). Growing evidence implicates elastin degradation and related host immune responses in the pathogenesis of AAA. Effective modeling of these processes is important for improving the understanding and treatment of this common disease. While previous AAA models have used elastin-disrupting techniques for aneurysm induction, the single use of elastase in many of these models does not account for the ongoing immune response against elastin and its degradation products (EDP). To model this ongoing immune response, we developed a model that sensitizes C57BL/6 mice to EDP by subcutaneously injecting EDP 7 days prior to aneurysm induction surgery. Aneurysm induction surgery proceeds with exposure of the abdominal aorta and application of topical elastase in the periaortic compartment, with the intention to expose elastin and EDP on the mouse aorta. With these moieties exposed, the animal should mount an immune response against these antigens to which it was previously sensitized. Repeat “booster” EDP injection 7 days later serves to propagate this immune response. Herein, we describe the protocol for the reliable creation of AAA in C57BL/6 mice via multiple sensitizations with EDP, followed by AAA induction surgery with perivascular elastase application. This model can be created quickly and at moderate cost, while inducing >30% aortic dilation in 82% of mice by video micrometry and 73% by ultrasound measurement, and >50% dilation in 64% by video micrometry and 27% by ultrasound, as measured at day 21 post-induction surgery. This technique is also associated with a 95% intraoperative survival rate.

Introduction

Abdominal aortic aneurysm (AAA) affects >35 million people worldwide, and 4–8% of men aged 65-801,2. Rupture, the most devastating complication of AAA, typically occurs when the mechanical wall stress of the aneurysm exceeds the strength of the aortic tissue, leading to massive hemorrhage and an overall mortality rate of 80−90%3,4. Current management is aimed at prevention, with ultrasound screening of high-risk individuals and surgical repair when the risk of rupture exceeds the risk of repair, typically at 5.5 cm in diameter

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Protocol

The animal protocols were approved by the Indiana University Institutional Animal Care and Use Committee (protocol number 24027). All studies followed the National Research Council’s Guide for the Care and Use of Laboratory Animals and were compliant with animal welfare Animal Research: Reporting of In Vivo Experiments (ARRIVE) guidelines. Male C57BL/6J wild-type mice were received at 12 weeks of age for use in the experimental protocol. Animals were allowed to acclimate for 7 days prior to the start of the protocol. Induction surgeries are performed 14 days after animals are received. Commercial lab rodent chow and water were supplied ad libitum, and mice were ....

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Results

A total of 28 male C57BL/6J mice have undergone this surgical protocol, with one perioperative mortality from an IVC injury (3.6%). There were three additional delayed mortalities, one of which was due to an incisional hernia and the other two of which were for unknown reasons, for a total 21-day survival of 85.7%. The average time from induction of anesthesia to the end of anesthetic administration was 46.4 ± 8.0 minutes. Sham-surgery mice underwent a nearly identical protocol to the aneurysm group, including initi.......

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Discussion

Since it was first described in rats in 1990, the elastase-based AAA model has been modified several times and used in various animals10,19,20,21,22,23. Some of the modifications include aortic cannulation, which is likely to increase mortality and introduce a traumatic component to the aortic pathology17

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Disclosures

The authors report no proprietary or commercial interest in any product mentioned or concept discussed in this article.

Acknowledgements

This work was supported by the Cryptic Masons Medical Research Foundation (Brownsburg, IN).

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Materials

List of materials used in this article
NameCompanyCatalog NumberComments
1 mL Tuberculin SyringesBecton Dickinson309628
10% Buffered Formalin PhosphateFisher ChemicalSF100-4
70% Isopropyl Alcohol SwabsCardinal HealthS-16183
Angled sharp forcepsDumontI I25 I-35
AutoClip 9mm (Skin stapler)MikRon427630
BetadineAtlantis Consumer Healthcare67618-155-16
Cotton gauze padDynarex Corporation3223
Cotton tipped applicatorPuritan Medical Products25-806 2WC
Elastase from porcine pancreas (≥4.0 units/mg protein)Sigma-AldrichE1250Stored in aqueous solution at 4 °C
Elastase from mouse pancreas (4.75 units/mg protein)Elastin Products Co., Inc.MS838Stored in aqueous solution (0.1 mg in 1 mL PBS)at 4 °C
ElastinMP Biomedicals101636Stored in aqueous solution at 4 °C
Enersight Desktop SoftwareLeicahttps://www.leica-microsystems.com/products/microscope-software/p/enersight/
Ethiqa XR (extended release buprenorphine)FidelisNDC 86084-100-30
Eye lubricantOptixCareOpx-4242
Flexacam i5 (Stereo)Leicahttps://www.leica-microsystems.com/products/microscope-cameras/p/12730536-leica/
Freund’s complete adjuvantMP Biomedicals642851Stored in aqueous solution at 4 °C
Freund’s incomplete adjuvantMP Biomedicals642861Stored in aqueous solution at 4 °C
Gloves (Sterile)Encore7824PF
Hypodermic Needles (23 G)Becton Dickinson305145
Hypodermic Needles (27 G)Becton Dickinson305109
Isospire (Isoflurane)Dechra Veterinary07-894-8943
Labeling TapeFisher Scientific15959
MasksDukal Corp.1540
Needle driverScanlan6006-36
Omnicon F/air Anesthetic Gas CanisterA.M. Bickford Inc.80120
Press'N SealGlad12587704417
RC2 Rodent Circuit ControllerVetEquip Incorporated922100
RNAlater Stabilization solutionThermoFisher ScientificAM7021
ScissorsFine Science Tools14005-12
Skin staples (clips)MikRon205016
Small Rodent Warmer HeaterStoelting53851
Small Rodent Warmer StageStoelting53812
Stereo Microscope M60Leicahttps://www.leica-microsystems.com/products/light-microscopes/stereo-microscopes/p/leica-m80/
Straight sharp forcepsDumontI I 25 I-30
SurgiSuite self-retaining magnetic retractor set and stageKent Scientific13-005-180
Tahoe Portable Oxygen ConcentratorVetland Medical595-2300A
Toothed forcepsFine Science Tools11043-08, 11042-08
Vevo 2100 UltrasoundFUJIFILM VisualsonicsEDU00496
Visorb polyglycolic acid sutureCP Medical397A
VWR AbsorbentsVWR95057-860

References

  1. Song P, He Y, Adeloye D, Zhu Y, Ye X, Yi Q, et al. The Global and Regional Prevalence of Abdominal Aortic Aneurysms: A Systematic Review and Modeling Analysis. Ann Surg. 2023;277(6):912-9.
  2. Kent KC. Clinical practice. Abdominal aortic aneurysms. N Engl J Med. 2014;371(22):2101-8.
  3. Macura KJ, Corl FM, Fishman EK, Bluemke DA. Pathogenesis in acute aortic syndromes: aortic aneurysm leak and rupture and traumatic aortic transection. AJR Am J Roentgenol. 2003;181(2):303-7.
  4. Hoornweg LL, Storm-Versloot MN, Ubbink DT, Koelemay MJ, Legemate DA, Balm R. Meta analysis on mortality of ruptured abdominal aortic aneurysms. Eur J Vasc Endovasc Surg. 2008;35(5):558-70.<....

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Tags

Murine AAA ModelElastic Lamellar MatrixElastin DegradationImmune ResponseAortic DilationC57BL 6 MiceAneurysm Induction
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