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Gene transduction into aortic wall using plasmid-loaded cationized gelatin hydrogel- coated polyester stent graft  Hongshan Zhong, MD, Osamu Matsui, MD,

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Presentation on theme: "Gene transduction into aortic wall using plasmid-loaded cationized gelatin hydrogel- coated polyester stent graft  Hongshan Zhong, MD, Osamu Matsui, MD,"— Presentation transcript:

1 Gene transduction into aortic wall using plasmid-loaded cationized gelatin hydrogel- coated polyester stent graft  Hongshan Zhong, MD, Osamu Matsui, MD, Ke Xu, MD, Takahiro Ogi, MD, Jun-ichiro Sanada, MD, Yasuo Okamoto, PhD, Yasuhiko Tabata, PhD, Yoh Takuwa, MD  Journal of Vascular Surgery  Volume 50, Issue 6, Pages (December 2009) DOI: /j.jvs Copyright © 2009 Society for Vascular Surgery Terms and Conditions

2 Fig 1 a, Partially-covered polyester stent graft; b, X-ray film of the rabbit that underwent the implantation of three CG hydrogel-coated partially-covered stent grafts (black arrows) in the aorta via right femoral artery. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

3 Fig 2 Whole mount X-gal staining of vessel segments and grafts. X-gal staining-positive blue dots and spots were revealed in both the aorta (a) and the neointima on the pCAGGS-LacZ-loaded graft (b). Parts of positive signals are indicated by black arrowheads. No blue dot was detected in the control aorta (c) or the graft (d). Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

4 Fig 3 X-gal staining and immunohistochemical staining of cryosections of the pCAGGS-LacZ-loaded stent graft implanted vessel segment. a, X-gal staining followed by fast-red nuclear counter staining showed blue positive dots (black arrows) in the neointima and the tissue ingrowth into the graft; b, Immunohistochemical staining using anti-α smooth muscle actin antibody. The α-smooth muscle actin-positive cells (brown color) were observed in the neointima, media, and vasa vasorum in the adventitia; c, Immunohistochemical staining using anti-macrophages (RbM2) antibody. RbM2-positive macrophages (brown color) were observed mainly in the neointima. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

5 Fig 4 Real-time reverse transcriptase polymerase chain reaction (RT-PCR) analysis of aortic segments and various organs for LacZ mRNA expression. LacZ mRNA (a) and β-actin mRNA as internal control (b). LacZ mRNA was detected only in the pCAGGS-LacZ-loaded, CG hydrogel-coated stent graft implanted vessel segment. No dissemination was observed to distal or proximal sections of the aorta or to remote organs. Positive control (PC) of pCAGGS-LacZ transfected Cos 7 cells; 1: Vessel segment with pCAGGS-LacZ-loaded stent graft; 2: Vessel segment with pCAGGS-loaded stent graft; 3: Vessel segment proximal to stent grafted (pCAGGS-LacZ) vessel segment; 4: Vessel segment distal to stent grafted (pCAGGS-LacZ) vessel segment; 5: Brain; 6: Heart; 7: Liver; and 8: Kidney. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

6 Fig 5 Time-dependent expression of LacZ in aortic segments and stent grafts after graft stent implantation. a, Whole mount X-gal staining of vessel segments and stent grafts at days 1, 3, and 7 after implantation of stent grafts loaded with pCAGGS-LacZ at the plasmid concentration of 10.0 mg/mL. Lower panel represents the magnified views of the blocked areas in the upper panel. Much greater numbers of X-gal staining-positive blue dots are observed in the vessel segment at day 1 after implantation compared with the segments at days 3 and 7. b, Quantitative real-time reverse transcriptase polymerase chain reaction (RT-PCR) analysis of LacZ mRNA expression in the aortic segment. The transduction efficiency at day 1 was significantly higher than at day 7 (*P < .05 by one-way analysis of variance (ANOVA) and Newman-Keuls multiple comparison test). Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

7 Fig 6 Dose-dependent expression of LacZ in aortic segments and stent grafts after graft stent implantation. a, Whole mount X-gal staining of vessel segments and stent grafts at day 1 after implantation of graft stents loaded with pCAGGS-LacZ at the plasmid concentrations of either 0.1, 1.0, or 10.0 mg/mL. Lower panel represents the magnified views of the blocked areas in the upper panel. Larger numbers of X-gal staining-positive blue dots are observed in the vessel segment with stent grafts loaded with 10.0 mg/mL plasmid solution compared with 1.0 and 0.1 mg/mL plasmid solutions. b, Quantitative real-time reverse transcriptase polymerase change reaction (RT-PCR) analysis of LacZ mRNA expression in the aortic segment. The transduction efficiency in 10.0 mg/mL plasmid was significantly higher than 1.0 and 0.1 mg/mL plasmid (*P < .05 vs 0.1 mg/mL loading concentration and f P < .01 vs 1.0 mg/mL loading concentration by one-way analysis of variance (ANOVA) and Newman-Keuls multiple comparison test). Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

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