| OBJECTIVE:Cryopreservation was an important technique in the study of skin graft. Cryoprotectants had been rapidly used to prevant the damage that tissue would get when it underwent freezing and thawing. This study comparatively analyzed the protection of three different cryoprotectants on the activity in cryopreserving rat skin, and provided the best one to lay the theoretical and experimental foundation for the clinical application of skin cryopreservant.METHODS:Healthy male Wistar rats were randomly divided into three experimental groups. Fresh skin were stored in the following three cryoprotectants as Trehalose/dimelhyl sulfoxide(group Ⅰ), Propanediol/dimelhyl sulfoxide(group Ⅱ), Acetamide/dimelhyl sulfoxide(group Ⅲ), and the treatment of each part within the cryoprotectant last for10,20,30,40,50,60min. The control group(group Ⅳ) used no cryoprotectant. Stored the experimental groups and control group at-26℃for24h,-80℃for14d after they were sealed in a plastic package and marked. Rewarmed the frozen rat skin quickly, then used the succinate dehydrogenase method to determinate the activity of rat skin.RESULTS:The activity of rat skin in group Ⅰ showed the best effect compared with other three groups. The results were analyzed statistically and the difference between the groups was statistically significant(P<0.05). The activity of rat skin increased with the time of saturating in the cryoprotectant and reached the maxism at30min. Then it decreased gradually and got less than the control group after60min.CONCLUSION:Trehalose/dimelhyl sulfoxide showed great advantage in protecting activity in cryopreserving rat skin compared with Propanediol/dimelhyl sulfoxide and Acetamide/dimelhyl sulfoxide. The protection could reach the best effect at the time of30min saturating in the cryoprotectant. |
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