Aortic wall cell proliferation via basic fibroblast growth factor gene transfer limits progression of experimental abdominal aortic aneurysm Katsuyuki.

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Presentation transcript:

Aortic wall cell proliferation via basic fibroblast growth factor gene transfer limits progression of experimental abdominal aortic aneurysm Katsuyuki Hoshina, MD, Hiroyuki Koyama, MD, Tetsuro Miyata, MD, PhD, Hiroshi Shigematsu, MD, PhD, Tsuyoshi Takato, MD, PhD, Ronald L. Dalman, MD, Hirokazu Nagawa, MD, PhD Journal of Vascular Surgery Volume 40, Issue 3, Pages 512-518 (September 2004) DOI: 10.1016/j.jvs.2004.06.018 Copyright © 2004 The Society for Vascular Surgery Terms and Conditions

Fig 1 In vivo gene transfer to aortic wall. a, A pair of electrode plates was positioned on the isolated abdominal aorta. b, After elastase infusion, plasmid solution was pooled for 20 minutes, followed by electroporation. Small bubbles appeared along the electrode during electric pulsation. Journal of Vascular Surgery 2004 40, 512-518DOI: (10.1016/j.jvs.2004.06.018) Copyright © 2004 The Society for Vascular Surgery Terms and Conditions

Fig 2 In vitro study for confirmation of plasmid transfer. Plasmid containing modified human basic fibroblast gene factor (bFGF) gene was transfected into cultured rat smooth muscle cells (SMCs). Culture medium of pCAcchbFGFcs23-treated SMCs (lanes 5-7) showed 2 forms of bFGF (18 and 22 kD), whereas medium samples of non-transfected SMCs (lanes 2-4) showed no band. Lane 1, molecular size marker. Journal of Vascular Surgery 2004 40, 512-518DOI: (10.1016/j.jvs.2004.06.018) Copyright © 2004 The Society for Vascular Surgery Terms and Conditions

Fig 3 In vivo expression of transfected gene. a, Frozen section of aortic wall 2 days after elastase infusion/electroporation of LacZ (×100). X-gal staining showed successful transfection (blue, arrow). b, Western blot analysis revealed that pCAcchbFGFcs23-treated samples (lanes 2-4) showed higher expression of bFGF than pCAZ3-treated samples (lanes 5-7). Lane 1, Molecular size marker. c, Densities of the bands on Western blot were quantified. Values are shown as mean ± SD. *P < .05). bFGF, pCAcchbFGFcs23-treated samples. Cont, pCAZ3-treated samples. Journal of Vascular Surgery 2004 40, 512-518DOI: (10.1016/j.jvs.2004.06.018) Copyright © 2004 The Society for Vascular Surgery Terms and Conditions

Fig 4 Maximum diameter of infrarenal aorta (a), and luminal area index (b). Average maximum diameter at day 7 was greater in bFGF group than in control group, whereas no difference was detected in luminal area index. Values are shown as mean ± SD. *P < .01, NS, no significant difference. Cont, control group; bFGF, bFGF group. Journal of Vascular Surgery 2004 40, 512-518DOI: (10.1016/j.jvs.2004.06.018) Copyright © 2004 The Society for Vascular Surgery Terms and Conditions

Fig 5 Photomicrographs of aneurysm wall from control group (a) and bFGF group (b) at 7 days after treatment (×200). c, Sections were immunostained with anti-α-smooth muscle actin antibody. α-Smooth muscle actin-positive cells stain brown. Medial smooth muscle cell density was evaluated by counting α-smooth muscle actin-positive cells in 4 high-power fields per section and averaging these values. d, Density of macrophages in aortic media was quantified in the same manner. e, CD31-positive cells on the luminal surface (endothelial cells) were counted. Values are shown as mean ± SD. *P < .01. NS, No significant difference. Cont, Control group; bFGF, bFGF group. Journal of Vascular Surgery 2004 40, 512-518DOI: (10.1016/j.jvs.2004.06.018) Copyright © 2004 The Society for Vascular Surgery Terms and Conditions

Fig 6 Photomicrographs of aneurysm wall from control group (a) and bFGF group (b) at 7 days after treatment (×400). Sections were immunostained with anti-bromodeoxyuridine (BrdU) antibody and counterstained with hematoxylin. BrdU-positive cells stain brown. Cell replication in the media was evaluated by counting BrdU-positive cells. Values are shown as mean ± SD. *P < .05. Cont, control group; bFGF, bFGF group. Journal of Vascular Surgery 2004 40, 512-518DOI: (10.1016/j.jvs.2004.06.018) Copyright © 2004 The Society for Vascular Surgery Terms and Conditions