Appropriate control of ex vivo gene therapy delivering basic fibroblast growth factor promotes successful and safe development of collateral vessels in.

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Appropriate control of ex vivo gene therapy delivering basic fibroblast growth factor promotes successful and safe development of collateral vessels in rabbit model of hind limb ischemia Shigeyuki Ishii, MD, Hiroyuki Koyama, MD, Tetsuro Miyata, MD, Seiji Nishikage, MD, Hirohumi Hamada, MD, Shin-Ichi Miyatake, MD, Hiroshi Shigematsu, MD Journal of Vascular Surgery Volume 39, Issue 3, Pages 629-638 (March 2004) DOI: 10.1016/j.jvs.2003.09.016

Fig 1 Experimental protocol for intra-arterial injection of various numbers of gene-transduced fibroblasts. Journal of Vascular Surgery 2004 39, 629-638DOI: (10.1016/j.jvs.2003.09.016)

Fig 2 Calf blood pressure ratio immediately after femoral artery excision, and immediately before, immediately after, and 28 days after cell administration. At 28 days there were significant differences between each group in line * and each group in line † (P < .01). There were no significant differences among the groups within each line. Journal of Vascular Surgery 2004 39, 629-638DOI: (10.1016/j.jvs.2003.09.016)

Fig 3 Selective internal iliac angiograms of rabbits at 28 days after injection of vehicle (a) and 2 × 105 (b), 1 × 106 (c), 5 × 106 (e), and 2.5 × 107 (e) AxCAMAssbFGF-tranduced fibroblasts. Arrow, internal iliac artery. f, Development of collateral vessels was quantified by angiographic score. There were significant differences between each group in line * and each group in line † (P < .05). There were no significant differences among groups within each line. Journal of Vascular Surgery 2004 39, 629-638DOI: (10.1016/j.jvs.2003.09.016)

Fig 4 a, Capillary density of left semimembranous muscle. b, In vivo blood flow of left internal iliac artery at rest. c, Maximum in vivo blood flow of left internal iliac artery. There were significant differences between each group in line * and each group in line † (P < .01 for capillary density; P < .05 for maximum blood flow). There were no significant differences between groups within each line. For blood flow at rest (b) there were significant differences between each group in line ‡ and the vehicle group (P < .05), but no significant differences among the groups within this line. Journal of Vascular Surgery 2004 39, 629-638DOI: (10.1016/j.jvs.2003.09.016)

Fig 5 Distribution of indium 111–labeled fibroblasts in organs and tissues at 5 hours after injection of 1 × 106 cells (a), 5 × 106 cells (b), and 2.5 × 107 cells (c) into the left internal iliac artery. Data are presented as percentage of radioactivity distributed in each tissue relative to total radioactivity of administered cells. d, Cell number of 111In-labeled fibroblasts in above-knee and below-knee muscles of left hind limb, lung, and liver at 5 hours after injection of 1 × 106, 5 × 106, and 2.5 × 107 cells into the left internal iliac artery. *P < .001. AK, Above-knee muscles; BK, below-knee muscles; Lt, left; Rt, right; Kd,; kidney; Ht, heart; Spl, spleen; Test, testis; NS, no significant difference. Journal of Vascular Surgery 2004 39, 629-638DOI: (10.1016/j.jvs.2003.09.016)

Fig 6 bFGF accumulation in left adductor muscle (a, b), lung (c), and liver (d) with Western blot technique. b, bFGF accumulation of control animals (21 days after femoral artery excision) was similar to that in vehicle-treated animals. PC, Positive control; vehicle, vehicle-treated sample; Cm, left adductor muscle sample from control animal; Clu, lung sample from control animal; Cli, liver sample from control animal; d, days after infection. Journal of Vascular Surgery 2004 39, 629-638DOI: (10.1016/j.jvs.2003.09.016)

Fig 7 a, Time course of systemic bFGF level measured with ELISA. b, Time course of anti-adenovirus antibody titer quantified with neutralizing test. Titers are shown as dilution ratio, and titers less than 1:4 were assigned a value of 1. * P < .0001 (5 × 106 cells vs 2.5 × 107 cells). pre, Preinjection; d, days after injection. Journal of Vascular Surgery 2004 39, 629-638DOI: (10.1016/j.jvs.2003.09.016)

Fig 10 Expression of luciferase in left above-knee (AK) muscles of rabbits. d, Days after second challenge. Journal of Vascular Surgery 2004 39, 629-638DOI: (10.1016/j.jvs.2003.09.016)