The role of short-term oxygen administration in the prevention of intimal hyperplasia Charu Lata, MBBS, Derrick Green, MD, Jing Wan, MD, Sabita Roy, PhD,

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The role of short-term oxygen administration in the prevention of intimal hyperplasia Charu Lata, MBBS, Derrick Green, MD, Jing Wan, MD, Sabita Roy, PhD, Steven M. Santilli, MD, PhD Journal of Vascular Surgery Volume 58, Issue 2, Pages 452-459 (August 2013) DOI: 10.1016/j.jvs.2012.11.041 Copyright © 2013 Society for Vascular Surgery Terms and Conditions

Fig 1 Creation of the arteriovenous fistula (AVF). A, Describes the normal anatomy of the iliac artery and vein and its surrounding vessels. B, After clamping the left common iliac artery and vein proximally and distally, the iliac vein was transected and the distal end oversewn. An arteriotomy was created in the common iliac artery, and the iliac vein was anastomosed to iliac artery in an end-to-side manner. The artery proximal to the fistula was considered the proximal artery, and the artery distal to the fistula was considered the distal artery. The vein immediately adjacent to the fistula was considered proximal vein and the vein distal to the fistula was the distal vein. IVC, Inferior vena cava. Journal of Vascular Surgery 2013 58, 452-459DOI: (10.1016/j.jvs.2012.11.041) Copyright © 2013 Society for Vascular Surgery Terms and Conditions

Fig 2 Effect of supplemental oxygen on intimal hyperplasia (IH) at the arteriovenous fistula (AVF). A, No change in IH with supplemental oxygen at day 1. B, No change in IH with supplemental oxygen at day 3. C, Supplemental oxygen shows a trend toward inhibiting IH at all sites at day 7. D, Supplemental oxygen significantly inhibits IH at the proximal vein and distal vein at day 21. E, Supplemental oxygen inhibits IH at all sites at day 42. F, Supplemental oxygen inhibits IH at all sites at day 90. Data are expressed as the ratio of intimal area (IA) to media area (MA). Data are expressed as mean ± standard error of the mean. P ≤ .05 was considered significant. Journal of Vascular Surgery 2013 58, 452-459DOI: (10.1016/j.jvs.2012.11.041) Copyright © 2013 Society for Vascular Surgery Terms and Conditions

Fig 3 Effect of supplemental oxygen on smooth muscle cell proliferation (SMCp). A, No significant proliferation was observed on day 1 in any treatment group. On day (B) 3 and (C) 7, the arteriovenous fistula (AVF) operation markedly increased the 5-bromo-2′-deoxyuridine (BrdU) staining, suggestive of proliferation at all sites. However, it was not significantly altered with oxygen supplementation. D, On day 21, the AVF operation markedly increased BrdU staining at all sites, and this proliferation was significantly decreased by oxygen supplementation at the site of the fistula and the distal vein. On day (E) 42 and (F) 90, there was no BrdU staining, suggesting absence of proliferation. This suggests that SMCp is a self-limiting event. Data are expressed as percent units (positively stained BrdU cells/all cells). Data are expressed as mean ± standard error of the mean. P ≤ .05 was considered significant. Journal of Vascular Surgery 2013 58, 452-459DOI: (10.1016/j.jvs.2012.11.041) Copyright © 2013 Society for Vascular Surgery Terms and Conditions

Fig 4 Intimal hyperplasia (IH) at the distal vein. A and B, A 90-day nonoxygen-treated rabbit (hematoxylin and eosin stain, original magnification, ×100, ×200). The yellow arrow represents the increased area of intima in a nonoxygen- treated vein. C and D, A 90-day oxygen-treated rabbit (hematoxylin and eosin, original magnification, ×100, ×200). Journal of Vascular Surgery 2013 58, 452-459DOI: (10.1016/j.jvs.2012.11.041) Copyright © 2013 Society for Vascular Surgery Terms and Conditions

Fig 5 Cellular proliferation at arteriovenous fistula (AVF). Immunoperoxidase staining of 5-bromo-2′-deoxyuridine(BrdU)-labeled cells. The black arrows identify darkly stained nuclei, which represent BrdU-postive staining cells. A and B, A 21-day nonoxygen-treated rabbit (biotinylated mouse anti-BrdU, original magnification, ×100, ×200). C and D, A 21-day oxygen-treated rabbit (biotinylated mouse anti-BrdU, original magnification, ×100, ×200). Journal of Vascular Surgery 2013 58, 452-459DOI: (10.1016/j.jvs.2012.11.041) Copyright © 2013 Society for Vascular Surgery Terms and Conditions