Preservation of rat limbs by hyperbaric carbon monoxide and oxygen

Naoyuki Hatayama,Munekazu Naito,Shuichi Hirai,Xiao-Kang Li,Hayato Terayama,Masahiro Itoh,Jun Araki,Masayuki Matsushita,Hiroki Yokota

doi:10.1038/s41598-018-25070-y

Abstract

Cold ischemia times ranging from <6 h to as long as 24 h are generally quoted as the limits for attempting the replantation of amputated extremities. In this study, we aimed to assess the effect of hyperbaric carbon monoxide (CO) and oxygen (O2) on rat limb preservation. Donor rat limbs were preserved in a chamber filled with hyperbaric CO and O2 for 3 days (CO + O2 3 days) or 7 days (CO + O2 7 days). Positive and negative control groups were created by using non-preserved limbs (NP) and limbs wrapped in saline-moistened gauze for 3 days (SMG 3 days), respectively. The survival rate of transplanted limbs at postoperative day 90 was 88% in the NP and 86% in the CO + O2 3 days. The corresponding survival rate was 50% in the CO + O2 7 days at postoperative day 90 but was 0% in the SMG 3 days at postoperative day 3. Muscle mass decreased in the CO + O2 3 days and CO + O2 7 days compared with the NP, but sciatic–tibial nerve conduction velocities did not differ. These results indicate that amputated extremities preservation with hyperbaric CO and O2 could extend the time limits of preservation, maintaining their viability for replantation.

Highlights

Replantation is the surgical reattachment by microsurgical techniques of a body part—most commonly a finger, a hand, or an arm—that has been completely cut from the body
Donor rat limbs were preserved in a chamber filled with Carbon monoxide (CO) and O2 for 3 days (CO + O2 3 days) or 7 days (CO + O2 7 days) at 4 °C under high pressure
Positive and negative control groups were created by using non-preserved limbs (NP group) and limbs wrapped in saline-moistened gauze (SMG) for 3 days at 4 °C (SMG 3 days), respectively

Summary

Introduction

Replantation is the surgical reattachment by microsurgical techniques of a body part—most commonly a finger, a hand, or an arm—that has been completely cut from the body. Given that accidents occur suddenly, and because time is needed to find a specialist to perform replantation, novel methods are needed to extend ischemia times when preserving amputated extremities for replantation[12]. Research has shown that these beneficial effects can somewhat prevent ischemia–reperfusion injury (IRI) following transplantation, with a stock solution of bubbling low-dose CO helping to preserve pig kidneys and rat livers[15,16]. We tried to preserve rat hearts in CO and O2 at high pressures and succeeded in preserving tissue for 24–48 h19. We aimed to assess the efficacy of using CO and O2 under high pressures to preserve rat limbs. We transplanted preserved rat hind limbs to syngeneic recipient rats in which the corresponding limbs had been removed

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Conclusion