Experimental study of the compound tissue preservation

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(1)The vascular compliance and the contractility of vascular smooth muscle of rat freshly amputated hindlimbs and hindlimbs preserved at 4ﾟC or-1ﾟC for 12,24,48, and 72 hours were evaluated using a fresh blood extracorporeal circulatory system. Vascular compliance after storage at -1ﾟC for 12 hours or longer decreased significantly from that of freshly amputated limbs (p<0.01). The contractility of vascular smooth muscle after storage at -1ﾟC for up to 24 hours were not different from that of freshly amputated limbs. However, it decreased significantly after 48 hours or longer. A comparison of these results to limbs Preserved at 4ﾟC suggested that the storage at -1ﾟC was inferior to that at 4ﾟC with respect to the vascular compliance, but was superior to the storage at 4ﾟC with respect to smooth muscle contractility. (2)The application of hypothermia to the replanted limb was observed. The mean arterial blood flow decreased significantly by 44% with cooling at 25ﾟC.The oxygen consumption decreased significantly by 28% of normothermia values. (3)Messenger RNA for GAPDH (glyceraldehyde 3-phosphate dehydrogenase) on the preserved anterior tibial muscle of rats were evaluated quantitatively using northen blot analyzes. The amputated hindlimbs were divided into 7 groups : 0,1,3,6,9 hours storage at room temperature (22ﾟC) and 6,9 hours storage at 4ﾟC.After a predetermined period of ischemia, each muscle was resected from the hindlimb and mRNA was analyzed. In the control group, the band for GAPDH was clearly detected. However, the bands were smeared in normothermic preserved groups. After 3 or more hours of storage at room temperature, mRNA content was significantly less than freshly amputated muscle. After up to 9 hours storage at 4ﾟC,mRNA did not decrease markedly. These results indicated that mRNA degradation of GAPDH correlate with muscle damage, since muscle damage was increased as preserved time passed.