Annette Ebner, PhD, David M. Poitz, PhD, Antje Augstein, PhD, Ruth H

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Functional, morphologic, and molecular characterization of cold storage injury Annette Ebner, PhD, David M. Poitz, PhD, Antje Augstein, PhD, Ruth H. Strasser, MD, PhD, Andreas Deussen, MD, PhD Journal of Vascular Surgery Volume 56, Issue 1, Pages 189-198.e3 (July 2012) DOI: 10.1016/j.jvs.2011.12.012 Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 1 Vessel function after cold storage at 4°C in an optimized storage solution. Control vessels (0 days) were stored for 2 hours. Cold storage for 2 days significantly impairs vascular function with respect to potassium-induced constriction and acetylcholine as well as nitroprusside-induced relaxation compared with control conditions. The error bars show the standard error of the mean. Journal of Vascular Surgery 2012 56, 189-198.e3DOI: (10.1016/j.jvs.2011.12.012) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 2 Contributions of different endothelium-derived mediators to acetylcholine (ACH)-dependent vessel relaxation: (A) maximum relaxation, (B) relaxation sensitivity (half-maximum effective dose [EC50]). Relaxation in mouse aorta vessel is mostly mediated via nitric oxide synthase signaling, because only a small residual relaxation is observed in the presence of L-NG-monomethyl arginine (L-NMMA). Inhibition of cyclooxygenase by indomethacin (INDO) does not alter maximum relaxation. In the combined presence of indomethacin and L-NMMA (I+L), no impairment of residual relaxation by cold storage was observed. The error bars show the standard error of the mean. Journal of Vascular Surgery 2012 56, 189-198.e3DOI: (10.1016/j.jvs.2011.12.012) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 3 Contributions of different potassium channel types to acetylcholine (ACH)-dependent vessel relaxation: Impairment of acetylcholine-dependent vessel relaxation was observed in the presence of the adenosine triphosphate-dependent potassium channel inhibitor glibenclamide (GC) but not in the combined presence of charybdotoxin and apamin (A+C), inhibitors of large and intermediate calcium-sensitive potassium channels. The error bars show the standard error of the mean. Journal of Vascular Surgery 2012 56, 189-198.e3DOI: (10.1016/j.jvs.2011.12.012) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 4 Acetylcholine-dependent vessel tone modulation. Left panel, Mouse aorta displays two distinct responses to acetylcholine (ACH). In low concentration (<10 to 6.5), ACH is able to relax a preconstricted vessel; in higher concentrations (>10 to 6), relaxation ceases and turns toward vasoconstriction. Right panel, ACH-induced maximum vasoconstriction (expressed as percentage of phenylephrine preconstriction) is impaired by 2 days of cold storage. This response is blocked by inhibitors of neuronal and inducible nitric oxide synthase (TRIM), thromboxane A2/prostaglandin H receptor (SQ29548), or cyclooxygenase (INDO). The error bars show the standard error of the mean. Journal of Vascular Surgery 2012 56, 189-198.e3DOI: (10.1016/j.jvs.2011.12.012) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 5 Staining with (top) picro-Sirius red and (middle) elastica van Giessen, and (bottom) α-actin immunohistochemistry of aorta cross-sections shows no loss or disruption of connective tissue (red in the top panel) and that elastic fiber (black-lilac in the middle panel) was induced by prolonged cold storage. Smooth muscle cell organization was not disturbed. Original magnification was 100× for overview pictures and 400× for zoom views. Journal of Vascular Surgery 2012 56, 189-198.e3DOI: (10.1016/j.jvs.2011.12.012) Copyright © 2012 Society for Vascular Surgery Terms and Conditions