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Murine abdominal aortic aneurysm model by orthotopic allograft transplantation of elastase-treated abdominal aorta  Zhenjie Liu, MD, PhD, Qiwei Wang,

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Presentation on theme: "Murine abdominal aortic aneurysm model by orthotopic allograft transplantation of elastase-treated abdominal aorta  Zhenjie Liu, MD, PhD, Qiwei Wang,"— Presentation transcript:

1 Murine abdominal aortic aneurysm model by orthotopic allograft transplantation of elastase-treated abdominal aorta  Zhenjie Liu, MD, PhD, Qiwei Wang, MS, Jun Ren, MS, Carmel Rebecca Assa, Stephanie Morgan, PhD, Jasmine Giles, BS, Qi Han, BS, Bo Liu, PhD  Journal of Vascular Surgery  Volume 62, Issue 6, Pages e2 (December 2015) DOI: /j.jvs Copyright © 2015 Society for Vascular Surgery Terms and Conditions

2 Fig 1 A, Schematic representation of aorta transplant model of murine abdominal aortic aneurysm (AAA) induced by elastase. Before transplantation, the donor's abdominal aorta was perfused with porcine elastase (0.45 U/mL, 10 minutes) under a pressure of 100 mm Hg. The donor's aorta was then harvested and anastomosed to the recipient abdominal aorta in an end-to-side fashion. B, Representative photograph of elastase-treated allograft 14 days after the transplantation procedure. C, van Gieson stain shows the aneurysm formation of the transplanted abdominal aorta and the medial elastin degradation of aorta from the donor 14 days after surgery. Scale bar = 200 μm. Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

3 Fig 2 Aortic expansion in the transplant model of elastase-induced abdominal aortic aneurysm (AAA) was similar to that in the conventional elastase model. A, Representative pictures show the aortic expansion 14 days after surgery from (left to right) the inactive elastase-treated group (IE), elastase-treated group (E), inactive elastase-treated transplant group (TIE), and elastase-treated transplant group (TE). B, The percentage increase of maximum AAA diameters of each group. AAA is defined as a percentage increase in aortic diameter greater than 100% (red dashed line). n = 5 in each 7-day group, n = 6 in each 14-day group. **P < .01; ***P < .001 (E vs IE, TE vs TIE). NS, Not significant. Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

4 Fig 3 Macrophage (CD68+ cells) and lymphocyte (CD3+ cells) infiltration in elastase-treated abdominal aortas (E) and transplanted elastase-treated abdominal aortas (Transplanted E) 7 days after surgery. Scale bar = 50 μm. Quantification of macrophage and lymphocyte infiltration in aneurysm tissues as identified by CD68 and CD3 stains, respectively, expressed as CD68+ or CD3+ cells/nuclei. NS, Not significant, n = 5. Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

5 Fig 4 Transplanted elastase-induced abdominal aortic aneurysm (AAA) had an inflammatory phenotype similar to that of the conventional elastase-treated AAA. At 14 days after surgery, the AAA tissues were harvested from different experimental groups: inactive elastase-treated (IE, n = 8), transplanted inactive elastase-treated (Transplanted IE, n = 6), elastase-treated (E, n = 6), and transplanted elastase-treated (Transplanted E, n = 7). Relative fold changes of mRNA levels were determined by real-time polymerase chain reaction (PCR). Data are represented as mean ± standard error of the mean. *P < .05 (E vs IE, transplanted E vs transplanted IE). CCL5, Chemokine (C-C motif) ligand 5; IFN-γ, interferon γ; MCP-1, monocyte chemoattractant protein 1; NS, not significant; TNF-α, tumor necrosis factor α. Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

6 Fig 5 Both recipient- and donor-derived macrophages are present in elastase-treated transplant grafts. Elastase-treated abdominal aorta segments were allografted from C57B6 mice into GFP+ C57B6 mice (A) and vice versa (B). Representative confocal immunofluorescence images for macrophage marker CD68 (red) and GFP (green) in allografts 7 days after transplantation. Nuclei stained with DAPI (blue). The yellow arrowheads indicate CD68 and GFP double positive cells. GFP, Green fluorescent protein; Lu, lumen; WT, wild type. Scale bar = 100 μm. C, Quantification of GFP+/CD68+ or GFP−/CD68+ cells in transplanted elastase-treated aneurysms, n = 5. Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

7 Supplementary Fig 1 (online only)
A, Kaplan-Meier survival curve shows the survival rate of the first 45 elastase-treated orthotopic abdominal aortic transplants, which were divided into three groups in sequence: 1 to 15, 16 to 30, and 31 to 45. AAA, Abdominal aortic aneurysm. B, Analysis of the death causes. The intraoperative mortality is 24.4%, mainly due to bleeding. Other postoperative complications of this procedure include thrombosis of the aorta, aneurysm rupture, hypovolemic shock, and unknown causes. Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

8 Supplementary Fig 2 (online only)
Hematoxylin and eosin (H&E) staining of arterial tissues 7 days after surgery from different groups: inactive elastase-treated abdominal aorta (IE), n = 4; elastase-treated abdominal aorta (E), n = 5; transplanted inactive elastase-treated abdominal aorta (TIE), n = 3; and transplanted elastase-treated abdominal aorta (TE), n = 3. van Gieson stain shows elastin layer degradation in representative treated arteries 7 days after surgery. Smooth muscle alpha actin (SMaA) immunohistochemical stain shows increased smooth muscle degradation of the elastase-treated abdominal aorta transplant, which is similar to elastase-treated aorta in the conventional model. Scale bar = 50 μm. Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

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