Use of Brilliant Blue FCF during vein graft preparation inhibits intimal hyperplasia Michael J. Osgood, MD, Kevin Sexton, MD, Igor Voskresensky, MD, Kyle.

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Use of Brilliant Blue FCF during vein graft preparation inhibits intimal hyperplasia Michael J. Osgood, MD, Kevin Sexton, MD, Igor Voskresensky, MD, Kyle Hocking, PhD, Jun Song, BA, Padmini Komalavilas, PhD, Colleen Brophy, MD, Joyce Cheung-Flynn, PhD Journal of Vascular Surgery Volume 64, Issue 2, Pages 471-478 (August 2016) DOI: 10.1016/j.jvs.2015.02.028 Copyright © 2016 Terms and Conditions

Fig 1 Brilliant Blue FCF (FCF) inhibited platelet-derived growth factor (PDGF)-mediated migration in rat vascular smooth muscle cells. Migration of A7r5 cells was measured in a scratch assay in six-well plates. Cells were pretreated without (control) or with FCF (25 and 50 μM) before stimulation with PDGF. A, Cells invading the scratch were counted after 24 and 48 hours. Results are reported as relative increase in cell number compared with the control (n ≥ 4). The range bars show the standard error of the mean. *P < .05. B, Photomicrographs of representative experiment at 0 (unstimulated) and 48 hours (original magnification ×40). Journal of Vascular Surgery 2016 64, 471-478DOI: (10.1016/j.jvs.2015.02.028) Copyright © 2016 Terms and Conditions

Fig 2 Brilliant Blue FCF (FCF) inhibited platelet-derived growth factor (PDGF)-mediated proliferation in rat vascular smooth muscle cells. Proliferation of A7r5 cells was measured using a tetrazolium dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in 96-well plates. Cells were pretreated without (control) or with different concentrations of FCF before PDGF stimulation. Each treatment was performed in at least six wells and averaged for each assay. Results are reported as relative increase in absorbance compared with the unstimulated control (n = 6). The range bars show the standard error of the mean. *P < .05, **P < .005, n.s., nonsignificant. Journal of Vascular Surgery 2016 64, 471-478DOI: (10.1016/j.jvs.2015.02.028) Copyright © 2016 Terms and Conditions

Fig 3 Brilliant Blue FCF (FCF) treatment during graft explantation reduced intimal hyperplasia in the rabbit carotid interposition graft model. The external jugular veins (EJVs) from rabbits were stored in heparinized PlasmaLyte (HP) without (control) or supplemented with 50 μM FCF during a 60-minute explantation period before implantation into the carotid artery. Grafts were harvested after 28 days and stained to visualize the elastic lamina. Intimal and medial thickness were measured. A, Representative grafts stained with Verhoeff-Van Gieson in control or FCF-treated animals. The boxed areas in the top row (original magnification ×20) are shown in the bottom row (original magnification ×40). Scale bar = 50 μm. L, Lumen; M, media. The arrows indicate internal elastic lamina, and the white lines indicate the intimal thickness. B, Intimal thickness and (C) intimal-to-medial thickness (I/M) ratio were determined (n = 13). The range bars show the standard error of the mean. *P < .05. Journal of Vascular Surgery 2016 64, 471-478DOI: (10.1016/j.jvs.2015.02.028) Copyright © 2016 Terms and Conditions

Fig 4 Brilliant blue FCF (FCF) pretreatment reduced the intimal thickness of human saphenous vein (HSV) in organ culture. HSV rings were left untreated (control) or pretreated with FCF (50 μM) for 2 hours before culture in Roswell Park Memorial Institute medium supplemented with 30% fetal bovine serum for 14 days. Veins were stained to visualize the elastic lamina, and intimal layer thickening was measured. A, Results are presented as the relative change in intimal thickness compared with preculture rings (n = 8). The range bars indicate the standard error of the mean. *P = .016. B, Representative Verhoeff-Van Gieson stained HSV. The white lines indicate the intimal thickness. Scale bar = 500 μm. L, Lumen; M, media. Journal of Vascular Surgery 2016 64, 471-478DOI: (10.1016/j.jvs.2015.02.028) Copyright © 2016 Terms and Conditions