Preservation of rat limbs by hyperbaric carbon monoxide and oxygen

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.

Application of Cryopreservation Technique in the Preservation of Rat Limbs

“…Although, in subsequent experiments, studies have found that after wrapping the limb with saline-moistened gauze, sealing it in a plastic bag, and keeping it on ice for 94 h, the nger can still be successfully replanted. However, in order to ensure the success of the operation, it is still recommended that the separated limbs be retransplanted within 12-24 h [4,5].…”

Different effects of partial pressure in a high-pressure gaseous mixture of carbon monoxide and oxygen for rat heart preservation

“…It was also shown that rat heart function after 24 h of high-pressure preservation using a mixture of CO and O 2 gases was almost the same as control hearts 14 . Moreover, we reported successful transplantation of a rat limb following 7 days of preservation under conditions of CO + O 2 (PCO:PO 2 = 4:3 atm) 15 . We also showed significantly lower blood levels of creatine phosphokinase in recipients who received limbs preserved in a chamber containing CO and O 2 gases for 18 h after wrapped in saline-moistened gauze for 6 h compared with limbs only wrapped in saline-moistened gauze for 24 h 15 .…”

“…Moreover, we reported successful transplantation of a rat limb following 7 days of preservation under conditions of CO + O 2 (PCO:PO 2 = 4:3 atm) 15 . We also showed significantly lower blood levels of creatine phosphokinase in recipients who received limbs preserved in a chamber containing CO and O 2 gases for 18 h after wrapped in saline-moistened gauze for 6 h compared with limbs only wrapped in saline-moistened gauze for 24 h 15 . It was also reported that hyperbaric CO and O 2 successfully preserved rat kidney grafts for 24 h by protecting tubular epithelial cells from apoptosis and inhibiting inflammation 16 .…”

High-Pressure Carbon Monoxide and Oxygen Mixture is Effective for Lung Preservation

“…We recently developed a high-pressure gas (HPG) preservation method using a mixture of CO and oxygen (O 2 ) and succeeded in resuscitating rat hearts after long-term preservation [15,16], revealing the utility of this method through histological analysis and metabolic evaluation [17]. Furthermore, this preservation method was reportedly useful for the preservation of rat kidneys [18] and limbs [19]. In addition, the lung is a solid organ similar to the kidney and liver, although it can be filled with gas from the inside, which is its decisively unique character.…”