Antisense basic fibroblast growth factor gene transfer reduces early intimal thickening in a rabbit femoral artery balloon injury model David G. Neschis,

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Antisense basic fibroblast growth factor gene transfer reduces early intimal thickening in a rabbit femoral artery balloon injury model David G. Neschis, MD, Shawn D. Safford, MD, Abigail K. Hanna, MD, Jonathan C. Fox, MD, PhD, Michael A. Golden, MD Journal of Vascular Surgery Volume 27, Issue 1, Pages 126-134 (January 1998) DOI: 10.1016/S0741-5214(98)70299-X Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 A, En face views of luminal surface of multiple discontinuous segments of a single rabbit superficial femoral artery treated with Ad.lacZ at 1 × 1010 pfu/ml via 20 minute local intraluminal incubation and stained with X-gal chromagen 4 days after balloon injury and vector delivery. These images demonstrate successful gene delivery and protein production along the full length of the treated vessel. Note segment in upper left image that demonstrates demarcation between treated and nontreated zones. Original magnification, ×7.5. B, Cross-sectional photomicrograph of a segment of rabbit superficial femoral artery treated with Ad.lacZ at 1 × 1010 pfu/ml via 20 minute local intraluminal incubation and stained with X-gal chromagen 4 days after balloon injury and vector delivery. Segments were then embedded, sectioned, and counterstained with nuclear-fast red. This image demonstrates greater than 90% gene transfer efficiency with gene delivery through the thickness of the media. Original magnification, ×125. Journal of Vascular Surgery 1998 27, 126-134DOI: (10.1016/S0741-5214(98)70299-X) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Hematoxylin and eosin–stained photomicrographs of rabbit superficial femoral arteries 14 days after balloon injury. A, Treated with Ad.lacZ at 1 × 1010 pfu/ml. B, Treated with PBS vehicle alone. C, Treated with Ad.ASbFGF at 1 × 1010 pfu/ml. All treatment solutions were delivered as a local intraluminal incubation for 20 minutes, placed immediately after balloon injury and brief PBS irrigation. These photomicrographs demonstrate a marked inhibition of neointimal thickening in the Ad.ASbFGF-treated vessels. Original magnification, ×125. Journal of Vascular Surgery 1998 27, 126-134DOI: (10.1016/S0741-5214(98)70299-X) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 I/M thickness ratios 14 days after balloon injury immediately followed by a 20-minute local incubation of treatment solution. Treatment groups included PBS vehicle alone, Ad.lacZ at 1 × 1010 pfu/ml, and Ad.ASbFGF at 1 × 1010 pfu/ml. Graph demonstrates significant reduction in intimal thickening in Ad.ASbFGF group compared with PBS and Ad.lacZ control groups (p = 0.004 and p < 0.0001, respectively). There was no significant difference in intimal thickening between the PBS and Ad.lacZ groups (p = 0.27). Journal of Vascular Surgery 1998 27, 126-134DOI: (10.1016/S0741-5214(98)70299-X) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 4 Indirect fluorescence microscopy of rabbit superficial femoral artery segments 4 days after balloon injury, treated with either Ad.lacZ at 1 × 1010 pfu/ml or Ad.ASbFGF at 1 × 1010 pfu/ml by local intraluminal incubation immediately after injury. A, C, and E, Red emission demonstrates propidium iodide counterstained nuclei (visualizing all nuclei present) of Ad.lacZ-treated, negative control, and Ad.ASbFGF-treated vessels, respectively. B, D, and F, Green emission visualizes FITC-labeled nuclei of cells undergoing apoptosis of Ad.lacZ-treated, negative control, and Ad.ASbFGF-treated vessels, respectively. Images demonstrate an increase in apoptotic cells in media of Ad.ASbFGF-treated vessel (F) compared with Ad.lacZ-treated vessels or negative control. Negative controls were of an Ad.ASbFGF-treated vessel stained in parallel with other sections but incubated in the absence of terminal deoxynucleotidyl transferase (TdT) enzyme. Original magnification, ×125. Journal of Vascular Surgery 1998 27, 126-134DOI: (10.1016/S0741-5214(98)70299-X) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions