Functional, morphologic, and molecular characterization of cold storage injury

• Published in: Journal of vascular surgery 2012-07, Vol.56 (1), p.189-198.e3
• Main Authors: Ebner, Annette, PhD, Poitz, David M., PhD, Augstein, Antje, PhD, Strasser, Ruth H., MD, PhD, Deussen, Andreas, MD, PhD
• Format: Article
• Language: English
• Subjects: Acetylcholine - pharmacology; Animals; Aorta - physiopathology; Biological and medical sciences; Cryopreservation - methods; Endothelium, Vascular - drug effects; Endothelium, Vascular - physiopathology; Gene Expression; Indomethacin - pharmacology; Male; Medical sciences; Mice; Mice, Inbred C57BL; Microarray Analysis; Muscle, Smooth, Vascular - drug effects; Muscle, Smooth, Vascular - physiopathology; Nitric Oxide Synthase - pharmacology; omega-N-Methylarginine - pharmacology; Potassium Chloride - pharmacology; Real-Time Polymerase Chain Reaction; RNA, Messenger - analysis; Signal Transduction; Statistics, Nonparametric; Surgery; Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases; Time Factors; Tissue Survival; Tissue Transplantation; Traumas. Diseases due to physical agents; Vascular injuries: limbs, aorta, vena cava; Vascular surgery: aorta, extremities, vena cava. Surgery of the lymphatic vessels
• ISSN: 0741-5214; 1097-6809
• DOI: 10.1016/j.jvs.2011.12.012

Objective Cold storage is used to preserve tissue for later transplantation. There is particular interest in prolonging cold storage time for transplantation purposes. To date, the mechanisms that contribute to vascular dysfunction in response to cold storage are poorly understood. The present study aims to characterize cold storage injury of blood vessels on functional and molecular levels. Methods To assess vessel function of mouse aorta, isometric force measurements were performed in a Mulvany myograph after cold storage at 4°C for various intervals. Morphologic changes were judged by histologic analysis of aortic cross-sections. To characterize cold storage-induced alterations on RNA levels, microarray analysis with subsequent polymerase chain reaction analysis was performed. Results Cold storage for 2 days revealed significant impairment of vessel function with respect to potassium-induced vessel tone development and acetylcholine-induced vessel relaxation. Detailed analysis of acetylcholine-mediated vascular response using specific pharmacologic blockers revealed that calcium-activated potassium channels seem to be impaired after 2 days of cold storage. At this point, no severe histologic changes (eg, elastic fiber disruption) were visible. RNA expression of 24 genes was significantly changed due to cold storage even after 2 hours. These include genes associated with vessel tone development (prostaglandin E3 receptor), cardiovascular function (adiponectin), electron transport chain (uncoupling protein 1), or calcium signaling (protein kinase A regulatory subunit 2b). Conclusions Long-term cold storage impairs vascular function, especially with respect to potassium signaling by calcium-dependent potassium channels. Microarray analysis confirmed impairment of pathways that are involved in calcium signaling and vascular function. Furthermore, various genes were significantly altered even after 2 hours, significantly before functional impairment was observed.