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      Development of a Novel Rabbit Model of Abdominal Aortic Aneurysm via a Combination of Periaortic Calcium Chloride and Elastase Incubation

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          Abstract

          Background

          The purpose of this study was to introduce a novel, simple and effective technique for creating a reliable rabbit model of abdominal aortic aneurysm (AAA) via a combination of periaortic calcium chloride (CaCl 2) and elastase incubation.

          Methods

          Forty-eight New Zealand white rabbits were divided into four groups. The AAA model was developed via a 20-minute periaortic incubation of CaCl 2 (0.5 mol/L) and elastase (1 Unit/µL) in a 1.5-cm aortic segment (Group CE). A single incubation of CaCl 2 (Group C) or elastase (Group E) and a sham operation group (Sham Group) were used for the controls. Diameter was measured by serial digital subtraction angiography imaging on days 5, 15 and 30. Animals were sacrificed on day 5 and day 30 for histopathological and immunohistochemical studies.

          Results

          All animals in Group CE developed aneurysm, with an average dilation ratio of 65.3%±8.9% on day 5, 86.5%±28.7% on day 15 and 203.6%±39.1% on day 30. No aneurysm was found in Group C, and only one aneurysm was seen on day 5 in Group E. Group CE exhibited less intima-media thickness, endothelial recovery, elastin and smooth muscle cell (SMC) content, but stronger expression of matrix metalloproteinase-2, matrix metalloproteinase-9 and RAM11 compared to the controls.

          Conclusions

          The novel rabbit model of AAA created by using a combination of periaortic CaCl 2 and elastase incubation is simple and effective to perform and is valuable for elucidating AAA mechanisms and therapeutic interventions in experimental studies.

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          Most cited references37

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          Inflammation and cellular immune responses in abdominal aortic aneurysms.

          Peter Libby, R. Mitchell, Koichi Shimizu (2006)
          Expansion and rupture of abdominal aortic aneurysms (AAA) result in high morbidity and mortality rates. Like stenotic atherosclerotic lesions, AAA accumulate inflammatory cells, but usually exhibit much more extensive medial damage. Leukocyte recruitment and expression of pro-inflammatory Th1 cytokines typically characterize early atherogenesis of any kind, and modulation of inflammatory mediators mutes atheroma formation in mice. However, the mechanistic differences between stenotic and aneurysmal manifestations of atherosclerosis remain unexplained. We recently showed that aortic allografts deficient in interferon-gamma (IFN-gamma) signaling developed AAA correlating with skewed Th2 cytokine environments, suggesting important regulatory roles for Th1/Th2 cytokine balance in modulating matrix remodeling and important implications for the pathophysiology of aortic aneurysm and atherosclerosis. Further probing of their distinct aspects of immune and inflammatory responses in vascular diseases should continue to shed new light on the pathophysiologic mechanisms that give rise to aneurysmal versus occlusive manifestations and atherosclerosis.
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            Matrix metalloproteinases 2 and 9 work in concert to produce aortic aneurysms.

            G Matthew Longo, Wanfen Xiong, Timothy C. Greiner … (2002)
            Matrix metalloproteinases (MMPs) 9 and 2 are increased in human abdominal aortic aneurysm (AAA) tissue, but their precise role and potential interaction remain unclear. Experimental induction of aortic aneurysms in mice genetically deficient in these peptidases could provide new insight into AAA pathogenesis. Mice deficient in the expression of MMP-9 (MMP-9KO) or MMP-2 (MMP-2KO) and their corresponding wild-type background mice (WT) underwent AAA induction by abluminal application of calcium chloride (CaCl(2)). No aneurysm formation was observed at 10 weeks after treatment in either the MMP-9KO or the MMP-2KO mice, whereas the corresponding WT mice showed an average 74% and 52% increase in aortic diameter, respectively. Reinfusion of competent macrophages from the corresponding WT strains into knockout mice resulted in reconstitution of AAA in MMP-9KO but not MMP-2KO mice. These findings suggest that macrophage-derived MMP-9 and mesenchymal cell MMP-2 are both required and work in concert to produce AAA.
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              Elastin degradation and calcification in an abdominal aorta injury model: role of matrix metalloproteinases.

              Barry C Starcher, Dan T. Simionescu, Dina M Basalyga … (2004)
              Elastin calcification is a widespread feature of vascular pathology, and circumstantial evidence exists for a correlation between elastin degradation and calcification. We hypothesized that matrix metalloproteinase (MMP)-mediated vascular remodeling plays a significant role in elastin calcification. In the present studies, we determined that short-term periadventitial treatment of the rat abdominal aorta with low concentrations of calcium chloride (CaCl2) induced chronic degeneration and calcification of vascular elastic fibers in the absence of aneurysm formation and inflammatory reactions. Furthermore, the rate of progression of calcification depended on the application method and concentration of CaCl2 applied periarterially. Initial calcium deposits, associated mainly with elastic fibers, were persistently accompanied by elastin degradation, disorganization of aortic extracellular matrix, and moderate levels of vascular cell apoptosis. Application of aluminum ions (known inhibitors of elastin degradation) before the CaCl2-mediated injury significantly reduced elastin calcification and abolished both extracellular matrix degradation and apoptosis. We also found that MMP-knockout mice were resistant to CaCl2-mediated aortic injury and did not develop elastin degeneration and calcification. Collectively, these data strongly indicate a correlation between MMP-mediated elastin degradation and vascular calcification.
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                Author and article information

                Contributors
                Vincenzo Lionetti: Role: Editor
                Journal
                Journal ID (nlm-ta): PLoS One
                Journal ID (iso-abbrev): PLoS ONE
                Journal ID (publisher-id): plos
                Journal ID (pmc): plosone
                Title: PLoS ONE
                Publisher: Public Library of Science (San Francisco, USA )
                ISSN (Electronic): 1932-6203
                Publication date Collection: 2013
                Publication date (Electronic): 2 July 2013
                Volume: 8
                Issue: 7
                Electronic Location Identifier: e68476
                Affiliations
                [1 ]Department of Radiology, The First Affiliated Hospital of China Medical University, Shenyang, China
                [2 ]Department of Interventional Radiology, The First Affiliated Hospital, Zhengzhou University, Zhengzhou, China
                [3 ]Key Laboratory of Diagnostic Imaging and Interventional Radiology of Liaoning Province, Shenyang, China
                [4 ]Department of Ultrasound, The First Affiliated Hospital of China Medical University, Shenyang, China
                Scuola Superiore Sant’Anna, Italy
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: YB HZ KX. Performed the experiments: YB. Analyzed the data: YB HZ ZZ XQ. Contributed reagents/materials/analysis tools: YB HZ KX ZZ XQ. Wrote the paper: YB HZ. Provided valuable comments during revision: HZ XQ YX LR. Contributed to writing the manuscript: YX LR.

                Article
                Publisher ID: PONE-D-12-39632
                DOI: 10.1371/journal.pone.0068476
                PMC ID: 3699498
                PubMed ID: 23844207
                SO-VID: 32c298f6-f519-4c4e-bb7a-39900acf0e9c
                Copyright statement: Copyright @ 2013
                License:

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                Date received : 11 December 2012
                Date accepted : 5 June 2013
                Page count
                Pages: 9
                Funding
                This work was supported by the National Natural Science Foundations of China (Grant No. 81000654, 81171443 and 81000620). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Subject: Research Article
                Subject: Biology
                Subject: Model Organisms
                Subject: Animal Models
                Subject: Medicine
                Subject: Anatomy and Physiology
                Subject: Cardiovascular System
                Subject: Circulatory Physiology
                Subject: Cardiovascular
                Subject: Acute Cardiovascular Problems
                Subject: Aortic Diseases
                Subject: Clinical Research Design
                Subject: Animal Models of Disease
                Subject: Surgery
                Subject: Cardiovascular Surgery

                ScienceOpen disciplines: Uncategorized
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                ScienceOpen disciplines: Uncategorized

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