Comparison of renal allograft fibrosis after transplantation from heart-beating and non-heart-beating donors

BACKGROUND: Renal transplants from non-heart-beating donors (NHBDs) yield acceptable function and allograft survival rates in the medium term. However, the long-term results are less certain and there is a paucity of information relating to the development of chronic allograft nephropathy. The aim of this study was to compare allograft fibrosis in kidneys transplanted from NHBDs and conventional heart-beating donors (HBDs). METHODS: A series of 37 NHBD and 75 HBD renal transplants were studied. Protocol renal transplant biopsies were performed at 6 and 12 months after transplantation. Biopsy sections were stained with Sirius red to demonstrate interstitial extracellular matrix. Renal allograft fibrosis was quantified using a computerized image analysis system. RESULTS: The mean first warm ischaemia time for kidneys from NHBDs was 24 min. A significant delay in graft function occurred in eight of 75 recipients in the HBD group and 31 of 37 in the NHBD group (P < 0.001). There were no significant differences in the level of allograft fibrosis between the two groups at any time point. CONCLUSION: Despite high rates of delayed graft function secondary to a prolonged warm ischaemia time, NHBD kidneys do not appear to be more susceptible to the development of renal allograft fibrosis. This study supports the growing body of evidence that kidneys from NHBDs are an acceptable alternative to those from HBDs.     

A case-control comparison of the results of renal transplantation from heart-beating and non-heart-beating donors.

INTRODUCTION: The decline in heart-beating brainstem dead organ donors has necessitated the search for other organ sources. In the field of renal transplantation one alternative source currently available, but little used, is that of kidneys from non-heart-beating donors (NHBD). Reticence to use NHBD kidneys is in part due to concerns over the effect that warm ischemic may have subsequent graft function. Presented here are the results of the NHBD renal transplants at the Leicester transplant unit, and compared with matched heart-beating donor transplants as a case control analysis. METHODS: In order to analyze any differences in graft performance between the two organ sources, the confounding effect of other variables known to influence the outcome of renal transplantation was minimized by matching NHBD and HBD transplants for the following criteria: donor age and sex, first or re-transplant, anastomosis and cold times, tissue match and PRA sensitisation. Transplant performance was assessed primarily by graft survival, the statistical evaluation of which was by log rank analysis of Kaplan-Meier curves. RESULTS: 72 NHBD and 192 HBD transplants were performed over an eight year period. Of the 192 HBD transplants, 105 matched one or more of the NHBD by the criteria outlined above, and thus constituted the control group for comparison. There was no significant difference in overall graft survival between the two groups. The 5 year survival for the NHBD was 73% compared with 65% for HBD kidneys. When death with a functioning graft is treated as censored data, then these figures become 75% and 81% respectively, again without statistical significance. CONCLUSION: NHBD kidneys are a valuable additional source of organs for transplantation, with long-term survival, comparable to transplants from HBD.     

Thrombospondin-1 mRNA expression in experimental kidney transplantation with heart-beating and non-heart-beating donors

BACKGROUND: Ischemia-reperfusion (IR) is a risk factor for delayed graft function, a clinical syndrome more frequently observed in non-heart-beating donors (NHBDs). Previous studies have reported that transforming growth factor-beta1 (TGF-beta1) and hypoxia-inducible factor-1alpha (HIF-1alpha) gene expression increase in the first few days after kidney transplant and that this increase in TGF-beta1 expression is lower in NHBD animals. The purpose of this study was to extend the gene profile analysis by characterizing TGF-beta1 activator thrombospondin-1 (TSP-1) and genes related to HIF-1alpha such as heme oxygenase-1 (HO-1), nitric oxide synthase-2 (NOS-2) and NOS-3. METHODS: The experimental pig model of kidney transplantation comprised heart-beating donors (HBDs, n=9) and NHBDs (n=22). Cortical biopsies were collected after anesthetic induction (baseline), after warm ischemia (WI), after cold ischemia (CI), after 1 hour of reperfusion (1R) and 5 days (5D) after transplant. TSP-1, HO-1, NOS-2 and NOS-3 mRNA expression was determined by real-time PCR. RESULTS: No change in expression of any of the genes analyzed was found during the transplant procedure (WI, CI, 1R) in HBD and NHBD cortical samples. TSP-1 mRNA was significantly increased at 5D in NHBD animals but unchanged in the HBD group. HO-1 was up-regulated in HBD (p<0.05) and NOS-2 mRNA was significantly increased in both groups (p<0.05). No difference in NOS-3 expression was observed at 5D. CONCLUSIONS: The increased TSP-1 expression in NHBDs may indicate a compensatory response to the reported diminished TGF-beta1 expression. The augmented NOS-2 and HO-1 expression in HBDs could have a positive effect on the recovery of kidney function.     

Outcome of kidney transplantation from nonheart-beating versus heart-beating cadaveric donors

BACKGROUND: This study aimed to assess outcomes of kidney transplants from nonheart-beating (NHB) compared with heart-beating (HB) cadaveric donors with meta-analytical techniques. METHODS: A literature search was performed for studies comparing kidney transplants from NHB vs. HB cadaveric donors between 1992 and 2005. The following outcomes were evaluated: warm and cold ischemia times, primary nonfunction, delayed graft function, length of hospital stay, acute graft rejection, patient and graft survival, and post-transplant serum creatinine. RESULTS: Eighteen comparative studies of 114,081 patients matched the selection criteria; 1,858 received kidney from NHB and 112,223 from HB donor. Warm ischemia time was significantly longer for the NHB group by 24 min (P<0.001). Cold ischemia time was similar for the two groups (P=0.97). The incidence of primary nonfunction and delayed graft function was 2.4 times (P<0.001) and 3.6 times (P<0.001) greater, respectively, in the NHB group. Length of hospital stay was longer for the NHB group by 4.6 days (P<0.001). The 6-month, 2-year, and 5-year patient survival were similar between the two groups. The incidence of acute rejection was similar between the two groups whereas the initial graft survival advantage in favor of the HB group diminished gradually over the course of time. There was no statistically significant difference between the two groups for the recipient serum creatinine levels at 3 and 12 months after transplantation. CONCLUSION: NHB donors carry the potential of expanding the cadaveric kidney pool. Although, transplants from NHB donors are associated with a greater incidence of early adverse events, long-term outcomes appear comparable with those of transplants from HB donors.     

Ablating the ischemia-reperfusion injury in non-heart-beating donor kidneys

BACKGROUND: The objective of this study was to determine if allopurinol (AL) and/or trifluoperazine (TFP) added to the Belzer machine preservation solution (MPS) improves the function of non-heart-beating donor (NHBD) canine kidneys. METHODS: Anesthetized canines underwent bilateral dissection of the renal vessels, obtaining baseline flow. After removing one kidney (heart-beating donor [HBD]), the dog was exsanguinated. After remaining in situ for 120 min (30-min warm ischemia time, 90-min cold ischemia time), the second kidney was removed (NHBD), flushed, biopsied, and weighed. The kidneys were machine-perfused separately for 20 hr, and pressure, flow, and resistance were measured serially. The kidneys were randomly assigned to a perfusate group (G): G1=MPS, G2=MPS+TFP, G3=MPS+AL, and G4=MPS+TFP+AL. Kidneys were implanted separately into a single recipient dog. Flow, resistance, and urine output were measured serially for 4 hr. Blood and urine samples and kidney biopsies were then obtained. All measurements were standardized to 100 g of kidney weight. RESULTS: HBD kidneys functioned better than NHBD kidneys in all groups, as expected. Although perfusate G1 was the most effective solution for HBD kidneys, the TFP additive (perfusate G2) more effectively reversed the vasospastic effects of ischemia/reperfusion for NHBD than the MPS solution (G1) with or without other additives. In HBD kidneys, the addition of AL resulted in the best creatinine clearance; however, AL was less effective than MPS alone in NHBD kidneys. TFP+AL together were completely ineffective in preserving renal function, regardless of whether the kidneys were from HBD or NHBD. CONCLUSIONS: MPS+TFP more effectively protected renal function against reperfusion injury in the NHBD than MPS alone, AL, or AL+TFP. AL exerts a salutary effect on creatinine clearance in HBD but not in the NHBD. The TFP and AL combination should not be used together with the MPS in machine preservation of kidneys.     

Extracorporeal membrane oxygenation support of donor abdominal organs in non-heart-beating donors

Both family consent and legal consent were required for organ/tissue donation from non-heart-beating donors (NHBD) in Taiwan. A district attorney had to come to the bedside to confirm the donor's asystole, confirm the family consent, and complete some legal documents before a legal consent was issued for organ donation. The resultant warm ischemic time would be unpredictably long and in fact precluded the organ donation from NHBD in Taiwan. We developed a method of using extracorporeal membrane oxygenation (ECMO) to maintain NHBD for a longer time and prevent warm ischemic injury of the donor abdominal organs. After ventilator disconnection in NHBD, phentolamine and heparin were injected and mannitol infusion was given. After the donor's asystole was confirmed by the electrocardiogram (EKG) strip recording, the ECMO support was set up through the right femoral veno-arterial route, an occlusion balloon catheter was inserted through the left femoral artery to occlude the thoracic aorta, and bilateral femoral arteries were ligated. Usually, the ECMO could begin within 10 min after the donor's asystole. The ECMO, combined with a cooler, provided cold oxygenated blood to the abdominal visceral organs, and prevented their warm ischemic injuries. Under the ECMO support (range: 45-70 min), eight renal grafts were procured from 4 NHBD. With the exception of the first two renal grafts with delayed function, all others had immediate function postoperatively and dialysis was no longer needed. In conclusion, by our ECMO technique, NHBD could be maintained for a longer time and the renal grafts had better immediate postoperative function than those reported by other methods.     

Innovative pulmonary preservation of non-heart-beating donor grafts in experimental lung transplantation.

OBJECTIVE: Lung transplantation is limited by scarcity of donor organs. Lung retrieval from non-heart-beating donors (NHBD) might have the potential to extend the donor pool and has been reported recently. However, no studies in NHBD exist using the novel approach of retrograde preservation with Perfadex solution. METHODS: Asystolic heparinized pigs (n = 5/group) were continuously ventilated for 90, 180 or 300 min of warm ischemia. Lungs were then retrogradely preserved with Perfadex and stored at 4 degrees C in inflation. After 3 h of additional cold ischemia, left lung transplantation was performed. Hemodynamics, pO(2)/F(i)O(2) and dynamic compliance were monitored for 5 h. Intrapulmonary lung water was determined by both global wet-to-dry lung weight ratio (W/D ratio) and standard stereological examination of relative volume fractions of intraalveolar edema. All results were compared to sham-operated controls and to lungs obtained from standard heart-beating donors after retrograde preservation with Perfadex and 27 h of cold ischemia. Statistics comprised ANOVA analysis with repeated measures and Mann-Whitney tests. RESULTS: No mortality was observed. During flush preservation of NHBD lungs, continuous elimination of blood clots via the pulmonary artery was observed. Oxygenation, compliance, intraalveolar edema fraction and W/D ratio were comparable between groups, whereas PVR was significantly lower in sham-controls. CONCLUSIONS: Use of NHBD lungs is feasible and results in similar postischemic outcome when compared to sham-controls and standard preservation procedures even after 5 h of pre-harvest warm ischemia. Especially, the NHBD with high-risk constellations for intravascular coagulation might benefit from retrograde preservation by elimination of thrombi from the pulmonary circulation. This innovative technique might also be considered in situations, where brain-dead organ donors become hemodynamically unstable prior to onset of organ harvest. Further trials with longer warm and cold ischemic periods are initiated to further elucidate this promising approach of donor pool expansion.     

乌司他丁对无心跳兔供肺的保护作用

目的 研究乌司他丁(UTI)对无心跳供者(NHBD)供肺的保护作用及其作用机制.方法 选取新两兰大白兔作为NHBD供肺离体冉灌注实验的供、受者,随机将兔分为A、B、C三组,每组各5对.A组为对照组,先使供者发生失血性休克并维持30 min,然后静脉注射氯化钾使心脏停跳,行胸外心脏按压以维持循环10 min后,原位冷却供肺,并经肺动脉灌注4℃的低钾右旋糖苷(LPD)液,取出供肺并冷保存5 h,最后将供肺与受者建立NHBD供肺离体再灌汴模型,供肺冉灌注时间为90 min;B组为UTI灌注组,供肺灌注时,LPD液中加入UTI(500 000 U/kg),其余处理同A组;C组为UTI预处理组,供者在休克期间静脉注射UTI(50 000 U/kg)行预处理,其余处理同B组.再灌注后1、30、60和90 min 4个时点,监测供肺的血氧分压(PO2)和气道峰压(PAwP).再灌注结束后,计算供肺组织湿/干重量比(W/D),并制备供肺组织匀浆,采用分光光度法和硫代巴比妥酸法测定髓过氧化物酶(MPO)和丙二醛(MDA)的活性;采用逆转录聚合酶链反应检测白细胞介素-8(IL-8)和细胞问粘附分子-1(ICAM-1)mRNA的表达水平;观察供肺病理组织学的变化.结果 再灌注后1、30、60和90 min,B组和C组的PO2、PAwP以及供肺W/D与A组比较,差异均有统计学意义(P<0.05),但B组与C组间的差异无统计学意义;再灌注结束后,B组和C组供肺组织匀浆中MPO和MDA的活性均显著低于A组(P<0.05),IL-8和ICAM-1mRNA的表达水平较A组显著下降(P<0.05);C组MPO和MDA的活性低于B组(P<0.05),IL-8和ICAM-1 mRNA的表达水平较B组显著下降(P<0.05);三组供肺组织均存在不同稗度的损伤,其中A组损伤最重,C组最轻.结论 灌注液中加入大剂量的UTI对NHBD供肺具有保护作用,尤其在NHBD休克期间注射UTI预处理.这可能与UTI能清除氧自山基,抑制炎症细胞冈子的释放、抑制中性粒细胞激活从而减轻缺血再灌注损伤有关.     

大白鼠无心跳供体肺移植模型的建立

目的　建立无心跳供体肺移植的模型 ,探讨无心跳供体肺应用于肺移植的可能性。方法　 60只健康SD大白鼠 ,随机分成 3组 :1组为有心跳供体组 ;2组为无心跳供体 热缺血 3 0min组 ;3组为无心跳供体 热缺血 60min组。供肺置于 4℃低钾右旋糖苷 (LPD)液中 4h。受体鼠行左侧开胸术和原位左肺移植术。术后维持辅助呼吸 1h ,阻断右肺门。结果　 2组与 1组相比 ,受体存活时间、移植后肺顺应性、超微结构、粒细胞浸润、气体交换等指标的差异均无显著性 (P >0 .0 5 )。3组与 1、2组相比差异有显著性 (P <0 .0 5 )。结论　成功建立了无心跳供体肺移植的实验模型 ,采用无心跳供体肺是肺移植一种安全而有效的方法。热缺血 3 0min大白鼠无心跳供体肺适于肺移植     

Pharmacologic graft protection without donor pretreatment in liver transplantation from non-heart-beating donors

BACKGROUND: Non-heart-beating donors (NHBDs) are considered potential sources of transplant organs in an effort to alleviate the problem of donor shortage in clinical liver transplantation. We investigated the possibility of pharmacologic protection of hepatic allograft function from NHBDs without donor pretreatment. METHODS: Orthotopic liver transplantation was performed using pigs. In donors, cardiac arrest was induced by stopping the respirator. Forty-five minutes after cessation of the respirator, the liver was flushed with cold lactated Ringer's solution including heparin and with the University of Wisconsin (UW) solution, and then preserved for 8 hr at 4 degrees C in the UW solution. The pigs were divided into two groups: a control group and a treated group. In the treated group, an endothelin antagonist TAK-044 was added to the UW solutions (10 mg/L), and TAK-044 (10 mg/kg body weight) and a platelet activating factor antagonist E5880 (0.3 mg/kg body weight) were also administered to the recipients. RESULTS: TAK-044 and E5880 treatment significantly increased the 7-day survival rate of the recipients (100% vs. 17%, P<0.05). In the treated group, portal venous pressure immediately after reperfusion of the graft was significantly lower than in the control group, and postoperative increase in serum concentrations of glutamic oxaloacetic transaminase and total bilirubin was attenuated. Moreover, the energy charge and adenosine triphosphate concentration of the liver were rapidly restored after reperfusion. CONCLUSIONS: Pharmacologic modulation with TAK-044 and E5880 avoiding donor pretreatment can improve the viability of hepatic allografts procured from NHBDs.     

Lung transplantation from out-of-hospital non-heart-beating lung donors. one-year experience and results.

There is currently no method for preservation and functional evaluation of clinical out-of-hospital non-heart-beating lung donors (NHBLD) that can be applied practically and systematically in clinical lung transplantation programs. A new method of preservation and functional evaluation of the lung has been developed in NHBLD that is based on the knowledge of various experimental studies. Initially, the viability of lungs harvested this way was proved from preliminary functional and histologic tests. In November 2002, we started using lung allografts from non-heart-beating donors. Five lung transplantations (4 bipulmonary and 1 unipulmonary) were performed successfully. The short and mid-term results have been excellent and all recipients are alive. We report our initial experience, which we hope will be of help to those involved in clinical lung transplantation programs worldwide.     

大白鼠无心跳供体肺移植的实验研究

目的　探讨不同热缺血时间对无心跳供体肺的组织结构和功能的影响及无心跳供体肺应用于肺移植的可能性。　方法　取 6 0只健康大白鼠 ,随机分成有心跳组、无心跳 缺血 30min组、无心跳 缺血 6 0min组 ,每组 10对 ,分别为供体和受体。有心跳组供体在处死的同时灌注 4℃低钾右旋糖苷液 ,无心跳 缺血 30min组、无心跳 缺血 6 0min组供体处死后维持辅助呼吸 ,分别放置室温中30和 6 0min ,再灌注低钾右旋糖苷液 ,供肺置于 4℃低钾右旋糖苷液中 4h。受体鼠行左侧开胸术和原位左肺移植术。术后维持辅助呼吸 1h ,经右侧进胸 ,阻断右肺门。　结果　无心跳 缺血 30min组存活时间均超过 30min ,肺顺应性为 0 .16 4 0± 0 0 0 4 9,动脉血氧分压 (85± 4 )mmHg ,动脉血二氧化碳分压 (41 9± 1 9)mmHg ,腺苷核苷酸总量 (75 8± 30 )mol/ g蛋白 ,超微结构改变为轻度的淤血和肺实质水肿 ,与有心跳组相比 ,差异均无显著意义 (P >0 0 5 )。无心跳 缺血 6 0min组有 4只 10min后心跳停止 ,3只 2 0min后心跳停止 ,与另两组相比 ,所测各项指标的差异有显著意义 (P <0 0 5 )。　结论　采用无心跳供体是增加供肺来源的一种安全而有效的方法 ,热缺血 30min大白鼠无心跳供体肺适于肺移植。