Effect of IGL-1, a new preservation solution, on kidney grafts (a pre-clinical study)

Ischemia-reperfusion injury conditions short-term and long-term graft function. The effects of the inversion of K⁺ and Na⁺ concentrations and substitution with polyethylene glycol for hydroxyethyl starch in University of Wisconsin (K-UW) solution were evaluated in isolated perfused rat kidneys and in autotransplanted pig kidneys. In the rat model kidneys were cold-stored for 24 h in K-UW or Na-UW or Na-PEG UW solutions (IGL-1 solution). Fractional sodium reabsorption and glomerular filtration rate was better in kidneys preserved in Na-UW and IGL-1 solution than those preserved in K-UW solution. In the pig model the left kidney was harvested and preserved in K-UW or IGL-1 solution for 24 h and then transplanted. In the autotransplanted pig model, kidneys preserved in IGL-1 solution showed a better function and a significant reduction of MHC class II expression, cellular apoptosis and interstitial fibrosis. In conclusion, kidneys preserved in IGL-1 solution tolerated ischemia/reperfusion injury better than those preserved in K-UW solution.     

IGL-1 solution in kidney transplantation: first multi-center study

Abstract:  IGL-1 solution is characterized by inversion of K+ and Na+ concentrations in the University Wisconsin (UW) solution and polyethylene glycol 35 (PEG 35) substitution for hydroxy ethyl starch. In this prospective study, 121 patients transplanted with kidneys preserved in IGL-1 solution were compared to 102 patients grafted with kidneys preserved in UW solution. Serum creatinine and creatinine clearance, delayed graft function (DGF) and rejection episodes, patient and graft survival were evaluated in the first post-transplant year. Groups were comparable regarding to donor and recipient characteristics. Median creatinine levels were significantly lower in IGL-1 group from day 6 to day 14 and it decreased more rapidly in the IGL-1 group (from day 4 to day 15: p < 0.05). Creatinine clearance values were usually higher in the IGL-1 group for the first 15 d. During the follow-up period serum creatinine concentrations were lower in IGL-1 group at one, three, six and 12 months after transplantation (p = 0.04; p = 0.06, p = 0.01 and p = 0.08, respectively) while creatinine clearance values were similar during the follow-up. No significant difference in DGF and rejection rates as well as in patient and graft survival was shown between the two groups. Kidneys preserved in IGL-1 solution showed to have the same function as kidneys preserved in UW solution.     

温度对WMO-1号液保存犬肾的影响

目的 探讨温度对ＷＭＯ－１号液保存犬肾的影响。方法 采用保存－自体移植模型,比较了不同温度条件下ＷＭＯ－１号液保存犬肾２４小时后的肾功能和形态学变化。结果 保存肾的损害程度与保存温度有关。１５℃保存２４小时不足以防止肾脏缺血损害。５－８℃组术后最大血肌酐值和平均血肌酐值均显著低于１－４℃组和９－１２℃组,形态学变化亦较轻。结论 ＷＭＯ－１号液保存犬肾的最适宜保存温度为５－８℃,保存温度过低或过高均     

临床肾脏保存2086例经验

目的：总结高渗枸橼酸盐腺嘌呤（HC－A）肾保存液2086例临床应用经验。方法：1980年8月至2000年12月连续2086例肾移植，供肾采用HC－A肾保存液保存。按冷缺血时间分组观察移植后血肌酐，移植肾存活率和平均存活时间。结果：冷缺血时间＜12h(573例）、12－24h（779例）和＞24h(734例）组移植肾平均存活时间分别为9．32、10．93和7．23年，后者与前两组比较差异均有显著性意义（P＜0．05）；移植后血肌酐、移植肾存活率在各组间差异无显著性意义（P＞0．05）。约3．16％的移植肾因保存问题出现肾功能异常。结论：应用HC－A肾保存液可取得较好的近期肾功能恢复和长期移植肾存活，冷缺血时间以24h内最佳。     

Pulmonary preservation with Bretscheider''s HTK and Celsior solution in minipigs

Background: Pulmonary preservation with high potassium/low oncotic pressure Euro-Collins (EC) solution is associated with endothelial dysfunction and reduced surfactant function. We compared two low potassium solutions, histidine-tryptophane-ketoglutarate (HTK) and Celsior, to EC in lung ischemia-reperfusion injury. Methods: In 19 minipigs, the left lung was perfused in situ with cold preservation solution (EC, n=6; HTK, n=6; Celsior, n=7). Reperfusion was started after 90 min of warm ischemia. The right pulmonary artery and main bronchus were clamped. Bronchoalveolar lavage (BAL) was obtained before ischemia and after 2 h of reperfusion. Surfactant activity was determined from the BAL in a pulsating bubble surfactometer. Results: Animals in the EC group survived 3.7±1.4 h. Six Celsior and five HTK treated animals survived the observation period of 7 h (P<0.001). Compliance of the reperfused lung deteriorated less in both Celsior and HTK groups (P<0.001). In EC and HTK animals, the pO₂/FiO₂ ratio was lower (P=0.002), and pulmonary vascular resistance was higher (P=0.02) than in Celsior animals. Surfactant function was impaired after reperfusion in all groups. Conclusions: Compared to EC, HTK solution showed moderate and Celsior distinct improvement of post-ischemic pulmonary function. However, surfactant function was not well preserved in any group.     

亚低温下肾保存的实验研究

为了进一步提高肾移植的水平，了解肾脏在低温保存下的病理改变及生化指标的变化，利用低温可控冰箱，以ＨＣＡ液作为灌洗及保存液，对兔肾进行不同温度（４℃、０℃，－２℃、－４℃）及不同时间（７２、９６、１２０小时）的保存，经对保存肾脏进行形态及功能方面观察，认为灌洗后静脉流出液ＬＤＨ浓度及光、电镜观察是肾保存质量观察的可靠指标。同时，在－２℃以下温度保存的肾皮质细胞在镜下可见有冰晶形成引起的空泡，它对肾脏细胞及结构产生严重的损害。     

The effect of IGL-1 preservation solution on outcome after kidney transplantation: A retrospective single-center analysis

Institut Georges Lopez-1 (IGL-1) solution is increasingly used for kidney preservation, although little information on outcomes is available. Outcomes of all deceased donor kidneys preserved by IGL-1, University of Wisconsin solution (UW), or histidine-tryptophan-ketoglutarate (HTK) and transplanted in our center (2000-2018) were analyzed. Multivariable analysis for delayed graft function (DGF), functional DGF, estimated glomerular filtration rate (eGFR, CKD-EPI equation), proteinuria, acute rejection, death-censored graft loss, and patient survival were performed. A double robust approach, consisting of propensity score weighting and correction for confounders, minimized the risk of bias. In total, 1943 transplants were included: 234 with IGL-1, 1046 with UW, and 663 with HTK. As IGL-1 was only introduced in 2014, a prespecified sensitivity analysis of 917 kidneys (2010-2018) was performed using the same statistical approach. After weighting, IGL-1 retained a higher proportion of kidneys donated after circulatory death (DCD). IGL-1 was not independently associated with any of the outcomes when compared to UW or HTK. Sensitivity analysis between 2010 and 2018 showed similar results. In this retrospective analysis, using robust methodology to reduce the risk of bias, IGL-1 preservation results in equal outcomes compared to UW or HTK, despite more DCD transplants in the IGL-1 group.     

Effect of prostaglandin E1 on preservation injury of canine liver grafts preserved in UW solution

This study investigated whether prostaglandin E₁ (PGE₁) could reduce hepatic injury to the liver graft caused by harvesting and 24-h preservation in University of Wisconsin (UW) solution in a canine model. The PGE₁-treated group was intravenously administered 0.5 g/kg per minute of PGE₁ for 30 min before harvesting, as well as a concentration of 1 mg/l PGE₁ in the washout and UW solutions. In both the PGE₁-treated and the control group, all recipients survived for 1 week or more after transplantation. Arterial ketone body ratio (AKBR) remained over 1.0 in the early postoperative period. The PGE₁ group showed significant reductions in guanase, GOT, and LDH during the early postoperative period compared to the untreated control group. Histological examination disclosed partial mitochondrial swelling, hepatocyte vacuolation, and necrosis in the control group, while such abnormalities were rarely seen in the PGE₁ group. These results suggest that PGE₁ can effectively reduce hepatic injury to liver grafts preserved in UW solution prior to transplantation.     

川芎嗪改善高渗枸橼酸盐嘌呤溶液保存犬肾的效果

目的探讨一定浓度的川芎嗪对低温保存的犬肾的影响。方法分别以2～4℃加入川芎嗪（终浓度为4mg／L）的新型高渗枸橼酸盐嘌呤溶液（HC-A Ⅱ液，实验组）、HC-A Ⅱ液（HC-AⅡ液组）及UW液（UW液组）灌注肾脏，然后进行组织病理学观察及细胞凋亡测定。将按上述方法保存48、72h的肾脏进行自体移植，术后观察血肌酐浓度、恢复正常的时间以及受者的存活情况。结果体外保存实验中，在保存时间不超过48h时，三个组的组织形态基本相似，但保存72h后，实验组和UW液组肾组织的病理改变明显轻于HC-A Ⅱ液组，细胞凋亡指数也明显低于HC-A Ⅱ液组（P〈0．05），实验组和UW液组的上述指标相比较，差异无统计学意义（P〉0．05）。经保存的肾脏自体移植后，单纯HC-A Ⅱ液保存72h者有2只犬（2／5）因肾功能衰竭死亡；不论是保存48h还是72h，采用含川芎嗪的HC-AⅡ 液和UW液保存者移植后血肌酐水平明显低于单纯HC-A Ⅱ液保存者（P〈0．05），其肾功能恢复正常的时间也较单纯HC-AⅡ液保存者缩短（P〈0．05）。结论HCA-Ⅱ液中加入一定浓度的川芎嗪能改善肾脏低温灌注保存的效果。     

A preliminary report of the HTK randomized multicenter study comparing kidney graft preservation with HTK and EuroCollins solutions

The main goal of transplantation is to restore good renal function and to improve the quality of life of thousands of dialysis patients, something which can only be achieved by providing them with well functioning grafts. Delayed renal allograft function is a serious problem. It is important to prevent this complication because it makes the diagnosis of acute rejection in the early postoperative period difficult, increases the necessity for diagnostic procedures, introduces dialysis treatments and prolongs hospital stay. The aetiology of delayed graft function (DGF) is multifactorial, and factors including donor management, technique used for organ procurement and preservation, age, anatomical variations in the graft, ischemia periods, use of cyclosporine A (CyA) or recipient immunological reactions have been implicated. Using different preservation solutions DGF rates vary from 30% to 60%. Recent clinical data have demonstrated better preservation and improved renal function posttransplant with HTK and University of Wisconsin (UW) solutions compared to EuroCollins solution. In a randomized multicenter study in collaboration with the Eurotransplant organ exchange organization, the efficacy of the HTK solution in renal transplantation was compared to EuroCollins and UW solutions in two parallel prospective randomized trials. The first preliminary results comparing HTK and EuroCollins solutions are reported here.     

生理盐水替代HTK保存液保存活体供肾的临床研究

目的 探讨生理盐水替代组氨酸-色氨酸-酮戊二酸(histidine-tryptophan-ketoglurate,HTK)保存液对活体移植肾功能恢复的作用及可行性.方法 亲属活体供肾肾移植的患者82例,按患者意愿分为生理盐水组(29例)和HTK组(53例).生理盐水组用生理盐水作为移植肾灌注液,HTK组用HTK保存液作...     

The impact of liver preservation in HTK and UW solution on microcirculation after liver transplantation

Severe microcirculatory disturbances due to endothelial cell damage and leukocyte adherence during reperfusion of transplanted livers are considered to contribute to early graft failure. Since the degree of reperfusion injury after liver transplantation depends on the length of preservation time and the solution used for preservation, the aim of our study was to assess three solutions with respect to microvascular perfusion and leukocyte adhesion. Therefore, rat livers were stored up to 24 h in Euro-Collins (EC), University of Wisconsin (UW), or histidin-tryphtophan-ketoglutarate (HTK) solutions prior to orthotopic transplantation. The livers were studied in situ 60 min postoperatively using intravital fluorescence video microscopy. Using simple syringe flushing (10 ml), sinusoidal perfusion decreased below 50% in EC preserved livers after 8 h preservation, in HTK preserved livers after 16 h preservation, and remained higher than 70% in livers preserved in UW up to 24 h. Permanent adhesion of leukocytes was increased more rapidly in organs after 1, 8, 16, and 24 h preservation in HTK (16%, 15%, 34%, and 49.7% ± 4.7%) compared to those preserved in UW (15%, 18%, 17%; and 32.7% ± 3.3%; P < 0.05). Using a 10-fold volumn of the organ weight of HTK solution during the harvesting procedure, with an 8 min equilibration period, sinusoidal perfusion (39.6 ± 4.7%) and leukocyte adhesion (42.7 ± 3.1%) were not improved after 24 h. In contrast, equilibration with a volumn of approximately 40-times the liver weight improved sinusoidal perfusion (70.8% ± 2.7%; P < 0.01) and leukocyte adhesion (24.9% ± 3.1%; P < 0.01) significantly. Thus, using HTK solution, simple flushing prior to long-term cold storage resulted in microcirculatory disturbances when compared to UW solution. Larger volumns of HTK solution with an additional equilibration period of 8 min, however, reduced leukocyte adhesion and improved sinusoidal perfusion to a similar degree as UW solution.     

SMO保存液对犬肾低温保存期间细胞凋亡和能量代谢的影响

目的研究SMO保存液对犬肾低温保存期间细胞凋亡和能量代谢的影响。方法建立犬离体肾脏单纯低温保存模型，根据保存液的不同分为3组，分别用0～4℃HTK保存液（HTK组）、UW保存液（UW组）和自制的SMO保存液（SMO组）对供肾进行灌注和保存；于犬肾保存24、48、72h后取各组的肾皮质标本，进行组织病理学、细胞凋亡和组织能量代谢的检测。结果犬肾保存48和72h时的组织病理学损害，SMO组比HTK组轻，但在各时间点与Uw组基本相似。犬肾保存24h时的细胞凋亡指数，SMO组与HTK组比较，差异无统计学意义（P〉0．05），保存48和72h后，SMO组显著低于HTK组（P〈0．05）；但SMO组与Uw组在各时点比较，差异均无统计学意义（P〉0．05）。犬肾保存24和48h时组织中Na^＋ -LK^＋ATP酶的活性，SMO组与HTK组比较，差异无统计学意义（P〉0．05），保存72h时，SMO组明显高于HTK组（P〈0．05）；而SMO组与Uw组在保存的各时点差异均无统计学意义（P〉0．05）。结论SMO保存液在减轻细胞形态学改变、减缓肾脏细胞凋亡和保持Na^＋ -LK^＋ATP酶活性方面显著优于HTK保存液，与UW保存液相当。     

Effect of different preservation solutions on adenine nucleotide content and metabolism in human kidney transplantation

Differences in purine metabolism produced by three preservation solutions were studied by determining the adenine nucleotide (ATP, ADP, AMP, and IMP) and nucleoside (adenosine, inosine, and hypoxanthine) levels in human kidney cortical biopsies. Forty kidney allografts were studied using University of Wisconsin (UW) solution (n=20), Euro-Collins (EC) solution (n=12), and modified EC solution with mannitol (M;n=8). No significant differences were found between the three solutions studied with regard to ATP, ADP, or AMP changes. The mean ATP level (nmol/mg prot±SEM) at the end of preservation in the UW group was 2.7±0.3 nmol/mg, in the EC group 3.8±0.7 nmol/mg, and in the M group 2.3±0.4 nmol/mg. ATP 30 min after reperfusion in the UW, EC, and M groups was 5.7±0.8 nmol/mg, 6.4±1.0 nmol/mg, and 4.6±0.5 nmol/mg, respectively. However, an important difference appeared in the catabolic products determined. Kidneys perfused with UW solution had a significantly higher level of adenosine (2.6±0.6 nmol/mg), inosine (11.8±2.2 nmol/mg), and hypoxanthine (18.1±2.1 nmol/mg) at hypoxanthine of cold storage than those perfused with EC (0.4±0.1 nmol/mg, 2.0±0.8 nmol/mg, and 7.1±1.4 nmol/mg) and M solutions (0.2±0.05 nmol/mg, 0.5±0.1 nmol/mg, and 5.2±0.6 nmol/mg; P<0.05). These levels returned to initial values 30 min postreperfusion and there were no differences with the EC or M solution groups at that time. Thus, the adenosine present in UW solution does not appear to be useful in recovering the adenine nucleotide pool at reperfusion. Moreover, it produces a marked increase in degradation products. Our findings do not support the beneficial metabolic effect of UW solution in terms of adenine nucleotide metabolism in comparison with simpler and less expensive preservation solutions like EC.     

新型器官保存液低温保存猪肾效果的研究

目的研究新型器官保存液Lifor液对猪肾脏的低温保存效果。方法 24只白色杂种猪,随机均分为Lifor液保存组和威斯康星大学保存液（UW液）组各12只,建立离体肾脏非循环灌注模型,供肾取出后分别以0~4℃的Lifor液和UW液灌注并低温保存,再根据保存时间随机分成2个亚组,分别为保存24h、48h组,然后行猪自体肾移植。比较两组肾脏低温保存结束后其病理学及肾皮质三磷腺苷（ATP）含量的改变,并观察自体肾移植后肾功能恢复情况。结果供肾离体保存24h、48h后,Lifor液组与UW液组的肾组织病理学改变基本一致,肾皮质ATP含量比较差异无统计学意义（P〉0.05）。移植后两组血清肌酐水平比较差异亦无统计学意义（P〉0.05）。结论 Lifor液低温保存肾脏的效果与UW液相当,且成本低廉,因此更具有临床应用前景。     

含饱和氢气肾保存液对大鼠肾脏冷缺血再灌注损伤的影响

目的 评价含饱和氢气肾保存液对大鼠肾脏冷缺血再灌注损伤的影响.方法 健康雄性Wistar大鼠24只,周龄8～10周,体重200～ 250 g,采用随机数字表法,将其随机分为3组(n=8):对照组(H1组)大鼠仅切除右肾;普通肾保存液组(H2组)大鼠采用冷缺血再灌注模型,用4℃普通HC-A肾保存液对左肾行冷灌注和冷保存;含饱和氢气肾保存液组(H3组)大鼠操作同H2组,灌注液及保存液换用自制的4℃含饱和氢气HC-A肾保存液.于再灌注24 h时抽取下腔静脉血样,测定血清BUN、Cr、TNF-α和IL-6浓度;切取左肾,测定肾组织MDA和8-羟基脱氧鸟苷(8-OHdG)含量,光镜下观察肾组织病理学结果.结果 与H1组相比,H2组和H3组大鼠血清BUN、Cr、TNF-α和IL-6浓度及肾组织MDA和8-OHdG含量均升高(P＜ 0.05);与H2组相比,H3组血清BUN、Cr、TNF-α和IL-6浓度及肾组织MDA和8-OHdG的含量均降低(P＜0.05).H1组肾组织形态结构未见明显异常,H2组肾小管损伤明显,H3组肾小管损伤较H2组减轻.结论 含饱和氢气肾保存液可明显减轻大鼠肾脏冷缺血再灌注损伤.     

Comparison of histidine-tryptophan-ketoglutarate (HTK) solution versus University of Wisconsin (UW) solution for organ preservation in human liver transplantation

Over a 30-month period, 60 patients (30 in each group) suffering from end-stage liver disease or primary hepatic malignancy and scheduled for liver transplantation were enrolled in a prospective, randomized study to compare two methods of liver preservation: histidinetryptophan-ketoglutarate (HTK) solution versus University of Wisconsin (UW) solution. Entry criteria for both groups were: age (18–65 years), elective surgery (transplantable or urgent category of the recipients), first transplantations and harvesting procedure performed by the same team. The parameters under investigation were the clinical and laboratory data preand post-transplantation, as well as follow-up data such as complications and survival. There were no significant differences in the two groups as far as the evaluation criteria were concerned, even when cold ischemia time was more than 15h (n=7). A slight, yet not significant, increase in late complications of the biliary anastomoses could be seen in the UW group. Hepatocellular injury (SGOT, SGPT, GLDH, lactate) appeared to be more marked in the HTK group. These results suggest that both HTK and UW solutions are appropriate for clinical use in liver transplantation, even if cold ischemia time is more than 15h.     

非HLA抗体相关肾移植排斥反应的研究进展

人类白细胞抗原（HLA）是由HLA基因复合体所编码的产物,定位于6号染色体短臂上,是异体免疫的主要靶点,但HLA抗体阳性并不能解释所有的肾移植排斥反应。非HLA抗体是同种异体肾移植中供者基因表达的产物,术中缺血-再灌注损伤、异体和自身免疫相互作用、细胞外囊泡的介导作用等都可以触发免疫系统反应,促使非HLA抗体产生。多项研究表明非HLA抗体是诱发排斥反应、影响肾移植结局的重要因素。因此,本文就肾移植非HLA抗体类型及形成机制进行综述,总结非HLA抗体相关肾移植排斥反应的研究进展,以期为肾移植术后非HLA抗体相关排斥反应的研究提供参考。     

Static cold storage preservation of ischemically damaged kidneys. a comparison between IGL-1 and UW solution.

Especially in damaged organs, adequate organ preservation is critically important to maintain viability. Institut Georges Lopez-1 (IGL-1) is a new preservation solution, with an extracellular sodium/potassium ratio and polyethylene glycol as a colloid. The influence of warm and cold ischemia was evaluated in a rat Lewis-Lewis transplant model with a follow up of 14 days. Eight groups of donation after cardiac death donor kidneys were studied with warm ischemia of 0 and 15 min followed by 0- or 24-h cold storage (CS) preservation in IGL-1 or UW-CSS. Blood was collected daily during the first week and at day 14. Recipients were placed in metabolic cages at day 4 and 14 after transplantation allowing urine collection and adequate measurement of glomerular filtration rate. Focussing on inflammation, reactive oxygen species production, proximal tubule damage, proteinuria, histology, and renal function after transplantation we could not show any relevant difference between IGL-1 and UW-CSS. Furthermore, the combination of 15-min warm ischemia and by 24-h cold ischemia did not result in life sustaining kidney function after transplantation, irrespective of the used solution. In the present experiment, static CS preservation of ischemically damaged rat kidneys in either IGL-1 or UW-CSS rendered equal results after transplantation.     

速尿对改善供心保存的作用

目的研究在冷缺血期供心保存液中速尿对改善鼠心保存效果的影响。方法Ｓｔ．Ｔｈｏｍａｓ液中加入速尿（５０ｍｇ／Ｌ）停搏并冷保存大鼠心脏６小时,利用离体鼠心非循环式Ｌａｎｇｅｎｄｏｒｆ灌流功能测定模型,测定心脏功能的恢复,检测心肌细胞ＡＴＰ和ＴＡＮ的含量等。结果加速尿组保存后的左心功能恢复明显优于不加速尿组,加速尿组保存后心肌细胞ＡＴＰ和ＴＡＮ的恢复率明显高于不加速尿组。结论保存液中的速尿能改善供心的保护效果