title: Improved preservation of rat hindlimbs with the University of Wisconsin solution and butanedione monoxime.
abstract: The purpose of this study was to compare the effects of the University of Wisconsin solution plus butanedione monoxime, the University of Wisconsin solution without butanedione monoxime, and saline on the preservation of muscle tissue. Forty-nine rat hindlimbs were amputated and replanted. The study population was subdivided into four groups according to flushing solution, storage, and replantation protocols. The limbs of the control group (n = 12) were flushed with 20 ml University of Wisconsin solution and immediately replanted onto the same rat. In the remaining three groups, the limbs were immersed in solution, stored in a refrigerator at 4 degrees C for 24 hours, and then replanted onto a fresh rat. The limbs in the no flushout group (n = 7) were placed into storage in cold saline solution without being flushed. The limbs in the University of Wisconsin solution group (n = 17) were flushed with 20 ml of University of Wisconsin solution prior to storage, and those in the University of Wisconsin solution plus butanedione monoxime group (n = 13) were flushed with 20 ml University of Wisconsin solution plus 20 mM butanedione monoxime. Limb survival rate was 100 percent for the control and University of Wisconsin solution plus butanedione monoxime groups, 87 percent for the University of Wisconsin solution group, and 71 percent for the no flushout group. Seven days after replantation, ATP levels were 71 percent of control in the University of Wisconsin solution plus butanedione monoxime group, 33 percent in the University of Wisconsin solution group, and 29 percent in the no flushout group. Tissue K+/Na+ ratio showed that the University of Wisconsin solution plus butanedione monoxime group maintained electrolyte balance, whereas the balance was significantly lowered in University of Wisconsin solution and no flushout groups. The University of Wisconsin solution plus butanedione monoxime limbs did not exhibit cell swelling, whereas total tissue water values for the University of Wisconsin solution and no flushout groups increased significantly. Serum creatinine kinase, measured 24 hours after replantation, was 120 percent of control in the University of Wisconsin solution plus butanedione monoxime group, 550 percent in the University of Wisconsin solution group, and 772 percent in the no flushout group. Limbs in the University of Wisconsin solution plus butanedione monoxime group had more flexible ankle joints and pliable muscle (i.e., less contracture) than those in the University of Wisconsin solution and no flushout groups. In conclusion, rat hindlimbs can be preserved hypothermically for 24 hours using the University of Wisconsin solution, the University of Wisconsin solution plus butanedione monoxime, or saline. However, the University of Wisconsin solution plus butanedione monoxime limbs had better ATP levels and less cellular injury after replantation. Based on these results, we believe that, biochemically, flushing and storage of muscle tissue in the University of Wisconsin solution plus butanedione monoxime are the most effective means of those studied for preserving composite tissue grafts for 24 hours.
authors.affiliation: Division of Plastic and Reconstructive Surgery, University of Wisconsin Hospital and Clinics, Madison, USA.
authors.forename: M A
authors.initials: MA
authors.lastname: Norden
authors.forename: V K
authors.initials: VK
authors.lastname: Rao
authors.forename: J H
authors.initials: JH
authors.lastname: Southard
mesh_terms: D000227:Adenine Nucleotides / Q000378:metabolism; D000241:Adenosine; D000493:Allopurinol; D000818:Animals; D002801:Cholinesterase Reactivators; D003080:Cold Temperature; D003402:Creatine Kinase / Q000378:metabolism; D003931:Diacetyl / Q000031:analogs & derivatives*; D005978:Glutathione; D006614:Hindlimb; D007328:Insulin; D008297:Male; D018482:Muscle, Skeletal / Q000378:metabolism; D019188:Organ Preservation Solutions; D011188:Potassium / Q000378:metabolism; D011887:Raffinose; D051381:Rats; D011917:Rats, Inbred Lew; D012092:Replantation; D012964:Sodium / Q000378:metabolism; D013997:Time Factors
chemical_list: D000227:Adenine Nucleotides; D002801:Cholinesterase Reactivators; D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; C004717:diacetylmonoxime; D000493:Allopurinol; D012964:Sodium; D003402:Creatine Kinase; D005978:Glutathione; D003931:Diacetyl; D000241:Adenosine; D011887:Raffinose; D011188:Potassium