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Objective To investigate the isolation and protective effect of a new islet purification solution (IPS)-Optiprep solution on the islet in mouse models. Methods The digested pancreatic islets were divided into the IPS and UW groups according to the islet volume. The pancreatic islets were isolated by the continuous gradient density centrifugation using IPS-Optiprep or UW-Optiprep solutions. The purification efficiency and isolated islet activity of purification solution were compared between two groups. The diabetic mouse models were successfully induced and randomly assigned into three groups. In the experimental group (n=10), the mice received pancreatic islet transplantation using islets isolated and purified by the IPS-Optiprep solution. In the control group (n=10), the mice underwent pancreatic islet transplantation using islets isolated and purified by the UW-Optiprep solution. In the sham surgery group (n=5), the mice merely underwent surgery without pancreatic islet transplantation. Postoperative blood glucose levels were detected and compared among three groups. The blood glucose levels of intraperitoneal glucose tolerance test at postoperative 21 d were statistically compared between the experimental and control groups. The cost of the preparation of two isolation solutions was also compared. Results Compared with the UW group, the islet equivalent (IEQ), islet purity, recovery rate and islet integrity were significantly higher in the IPS group. Islet morphological observation revealed that the islet membrane was complete and the islet diameter in the IPS group was considerably larger than that in the UW group. The activity of purified islets in the UW group was significantly higher than that in the IPS group [(88±5)% vs. (84±3)%, P<0.01]. Compared with the UW-Optiprep solution, identical in vivo islet function was obtained in the IPS-Optiprep solution.The cost of IPS-Optiprep solution was significantly less than that of the UW-Optiprep solution. Conclusions The new IPS-Optiprep solution yields higher islet isolation efficiency, purification, integrity and recovery rate and significantly reduces the purification cost compared with the UW-Optiprep solution. Nevertheless, IPS-Optiprep solution exerts a less protective effect on the activity of islet cells, which is probably correlated with the high islet integrity and the endotoxin in the IPS-Optiprep solution.
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Objective To explore the application of UW solution in the replantation of the rat limb,and to explore a simple method for the ideal preservation of the amputated limbs.Methods From September,2017 to Janurary,2018,one hundred and twenty healthy adult male SD rats,weight 320-350 g,were randomly divided into 8 groups:A,B,C,D,E,F,G,H group.The isolated limbs of the control group A,C,E,G were cryopreserved in refrigerator at 4 ℃,while those of the experiment group B,D,F,H were preserved under the same condition after being perfused with UW solution.All the severed limbs were replanted back to the rats after different time length of cry opreservation:A,B 12 hours,C,D 20 hours,E,F 28 hours and G,H 36 hours.During the operations the femoral artery and venous patency were detected immediately after anastomosis.And after the operations the survival rates of limb replantation were observed.On the third day after the limb replantation,the tibial posterior muscle tissue was examined for pathology.The cell areas in pathological photographs were compared with Image analysis software Image-pro Plus 6.0 and the ultra structural changes of skeletal muscle tissue were observed.Results The femoral artery and venous patency in experiment group A,C,E and control group B,D,F were all 100 percent.However,in the group G,H the severed limb vessels changed obviously,the patency dropped markedly and the survival rates of limb replantation turned out to be 0.The experiment groups perfused with UW solutions,had relatively higher survival rates compared with the control groups (12 h:93.33％ vs.53.33％,20 h:80.00％ vs.40.00％,28 h:53.33％ vs.13.33％) and the skeletal muscles edema was lighter than that of the control group,with a larger proportion of cell areas(12 h:69.49±8.57 vs.44.75±6.34,20 h:56.34±7.73 vs.36.69±5.74,28 h:45.76±8.23 vs.31.49±7.72).There was a significantly difference (P＜0.05).Electron microscope observation showed that the experiment group bore a lighter damage of myofibril,interstitial tissue and mitochondria compared with the control group.Conclusion The combination of UW solution and cryopreservation of organs is an ideal and proper way to preserve the amputation limb,to extend the time limit for replantation,and to achieve a better recovery effect of postoperative limb function.Besides,UW solution is easy to be promoted in the treatment of isolated limb in clinical work.
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Objective To assess the effect of intraarterial washout with UW solution and tanshinone Ⅱ A for skeletal muscle preservation during ischemia and reperfusion.Methods Ischemic rat limbs were perfused with UW solution or UW solution plus tanshinone Ⅱ A (0.05,0.1 and 0.2 mg/ml) for 0.5 h before reperfusion; controls received no perfusion.Serum CPK,LDH,and AST were measured pre-ischemia and after reperfusion (2 h,4 h,and 6 h).Muscle water content,MDA,SOD,ATPase were assessed pre-reperfusion and after 6 h reperfusion.ICAM-1 was detected after 6 h reperfusion.Results Intraarterial washout with UW and UW + T could inhibit the express of ICAM-1 in skeletal muscle.The serum levels of CPK,AST,and LDH in UW group were significant lower than those in control group after 2 h of reperfusion,but no difference was observed between UW group and control group after 4 h and 6 h of reperfusion.After 4 h of ischemia,there were significant differences in water content,MDA,SOD,and ATPase in skeletal muscle between UW group and control group,but no difference was found after 6 h of reperfusion.In contrast,all parameters of laboratory test and biochemical analyzing in UW + T(0.05,0.10,and 0.20 mg/ml) groups were significantly different from those in control group at corresponding reperfusion period.Conclusion Intraarterial washout with UW solution is effective in preserving skeletal muscle integrity against I/R insult.Tanshinone Ⅱ A as a beneficial adjunct to UW solution improves the protective effect of UW solution for ischemic skeletal muscle.To better preserve ischemic skeletal muscle,an appropriate dose of tanshinone Ⅱ A (0.1 or 0.2 mg/ml)added to UW solution is required.
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Objective To determine the effect of the modified UW (University of Wisconsin)solution in the treatment of acute renal tubular necrosis in newborn swine. Methods Ten one-week-old newborn swine were used to establish the animal model of acute renal tubular necrosis by clamping their renal arteries,and were divided into two groups: the model group( n = 5 ) and the treatment group ( n = 5 ) in which fructose diphosphate sodium UW solution was used. Sham surgery was performed on other five swine, which were used as the sham group. At 12 h,l d,2 d,3 d and 7 d after the operation,the urine volume,urine protein,blood urea nitrogen(BUN) and creatinine(Cr) were determined. At 12 h ,24 h and 7 d after the operation ,renal pathological examination was conducted. Results The renal pathological examination and the blood biochemistry tests showed that the animal model was successful. BUN and Cr in the model group and the treatment group were significantly higher than those in the sham group at 12 h after operation(P ＜0. 05) ,and they arrived at their peak values at 2 d after operation,showed remarkable decline at 7 d,especially in treatment group,and returned to the level of the sham group. The urine protein in the model group and treatment group were higher than those in the sham group at various times(P ＜0.05 or P ＜0.01) and it peaked at 1 d after operation,then declining gradually,especially in the treatment group. Compared with the sham group,there were a significant decrease in the urine volume at various times in the model group(P ＜0. 05 or P ＜0. 01 ) ,while in the treatment group,the decrease in the urine volume were significant only at 12 h, 1 d and 2 d( P ＜ 0. 05 ) ,and turned insignificant at 3 d and 7 d. The pathological examination showed that the pathological changes in the treatment group were significantly milder than those in the model group. Conclusion The modified UW solution is effective in reducing the acute renal tubular necrosis in newborn swine.
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Objective To compare F-Z solution with UW solution in preserving efficacy at cell level. Methods The LDH content in supernatant of endothelial cells after preservation-rewarming to determine the injury after preservation-rewarming. MTT was used to determine the activity of endo-thelial cells after preservation-rewarming. Results After being preserved for 24 h, the LDH content in supernatant of endothelial cells in UW group was not significantly different from that in F-Z group.After preservation for 48 h, there was no marked difference in the activity of endothelial cells between the 2 groups. Conclusion F-Z solution and UW solution have the same effect in reducing injury and maintaining activity of endothelial cells after preservation-rewarming at cell level.
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Objective To compare the efficacy of liver preservation in SX-1 solution,HC-A solution and UW solution in the two-cuff technique orthotopic liver transplantation model in rats.Methods We confected SX-1 solution using of aseptic technique and built up two-cuff technique orthotopie liver transplantation model in rats.The SX-1 solution,UW solution and HC-A solution preserved rats livers in 2 h,8 h and 24 h,then went orthotopic liver transplantation.The various hepatic indices were determined when the rats survived for 6 hours after the operation.Results For the ALT,AST,the SX-1 solution group(2 h,8 h,24 h)and UW solution group went up synchronously,there was no statistical significance,(P＞0.05).Compared with HC-A solution group the SX-1 solution group (2 h,8 h,24 h)has already shown the statistical signifcance(P＜0.05).For the LDH,the SX-1 solution group (2 h,8 h,24 h)and HC-A solution group went up synchronously,there was no statistical significance(P＞0.05).Compared with UW solution group the SX-1 solution group(2 h,8 h,24 h)has already shown the statistical significance(P＜0.05).For the number of liver with bile production,each group didn1 show the statistical significance (P＜0.05).The time of preserving liver increased with the number of liver with bile production reducing.Conclusions Confirmed by the orthotopie liver transplantation model in rat,the SX-1 solution is the same as the UW solution in the indices of hepatic enzymologists.exceeding the HC-A solution.The SX-1 solution is the same as HC-A solution and UW solution in the number of liver with bile production.
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Organ preservation is companied with ischemia,hypothermic injury and reperfusion injury.This article describe basic organ preserve method,developing history,current state of organ,and several research problem.
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PURPOSE: In living donor partial liver transplantation, the organ cold preservation time is relatively short, thereby, Euro-Collins (EC) solution could be used with comparable result to University of Wisconsin (UW) solution. METHODS: About 70% partial liver was harvested and reimplanted after 3.5 hours of cold preservation with EC or UW solution in same mongrel dog weighing about 25 kg. In 5 cases EC solutions were used, and UW solutions were used in another 5 cases. Blood sample and liver biopsy specimens were taken just before donor hepatectomy, postreperfusion 1 hour, 7 days and 14 days after operation. Serum aspartate aminotransferase (AST), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) were monitored, and morphologic grading of liver injury was performed as Spiegel et al. reported in 1999. RESULTS: Total ischemic time was 4 hours and 27 minutes in average. At 1 hour after reperfusion, there were no significant differences in liver enzyme and morphological score between the UW solution group and the EC group. Survival rate was similar in two groups; 2 dongs were alive at postoperative 7th day, and 1 dog was alive at postoperative 14th day in each group. CONCLUSION: In living donor partial liver transplantation where both operations could be performed simultaneously with relatively short graft preservation time, EC solution could be used with comparable results to UW solution.
Subject(s)
Animals , Dogs , Humans , Alkaline Phosphatase , Aspartate Aminotransferases , Biopsy , Hepatectomy , L-Lactate Dehydrogenase , Liver Transplantation , Liver , Living Donors , Pathology , Reperfusion , Survival Rate , Tissue Donors , Transplants , WisconsinABSTRACT
The purposes of initial preservation solution are rapid cooling of the organ and washing out the intravascular components. After the introduction of University of Wisconsin (UW) solution, it has become the standard organ preservation solution in liver transplantation. However, due to several problems such as high viscosity and potassium concentration, let alone its cost, there has been several attempts to use UW solution in combination with other preservation solutions for initial perfusion of the graft during the organ harvest procedure. In order to evaluate the efficacy of Ringers' Lactated (RL) solution as an initial perfusion solution, we performed orthotopic hepatic allografts on dogs. In 4 dogs, UW solution was used as the initial perfusion solution and in the other 4, RL solution was used. After initial perfusion, UW solution was used successively for the preservation of the graft. All harvested grafts were stored in UW solution. Average cold ischemic time was 163.5 minutes for RL group and 159 minutes for UW group. The two groups were compared in terms of hematologic and biochemical markers before and after the transplantation. The extent of graft injury during cold ischemia and after reperfusion was also compared directly under light and electron microscope. There was no difference in cold and warm ischemic time, anhepatic time, and total operation time between the two groups. The groups did not differ in liver function test, complete blood count and coagulation profile. Histologic exams also showed similar changes between the two groups. In conclusion, RL solution can be used as the initial perfusion solution in hepatic transplantation.