Effects of warm ischemia and cryopreservation on cartilage viability of tracheal allografts

Background. For clinical use of a cryopreserved tracheal allograft, it is important to evaluate cartilage viability. We assessed cell viability of the cartilage in a cryopreserved tracheal allograft by measurement of (Na2SO4)-S-35 incorporation. We also investigated the effects of warm ischemic time on tracheal cartilage viability. Methods. The tracheas from Lewis rats were harvested and preserved at different warm ischemic times from cardiac death to preservation (0, 1, 2,4,6,9, and 12 hours, each group n = 8). The cartilage was labeled with 4 mu Ci/mL of (Na2SO4)-S-35. The specimen was hydrolyzed in 0.5 mol/L NaOH, and a solution of the extracts was then counted by liquid scintillation counter. Tracheas were transplanted into Brown Norway rats. Results. (35)Sulfur incorporation in the cartilage decreased as warm ischemic time increased. In addition, (35)Sulfur incorporation decreased from 76% to 67% after cryopreservation. Histologic examinations of the normal tracheal cartilage before preservation and after thawing were done in all the groups. After transplantation, the cartilage had severe fibrous changes, and its layer was almost nonobservable in the 9- and 12-hour groups. Conclusions. The viability of the tracheal cartilage decreased with warm ischemic time and from 76% to 67% after cryopreservation. In the rat tracheal transplantation model, a cryopreserved tracheal allotransplant could be done safely with a graft that was cryopreserved within 6 hours of warm ischemic time. (Ann Thorac Surg 2000;70:1876-9) (C) 2000 by The Society of Thoracic Surgeons.