The mode of death in the non-heart-beating donor has an impact on lung graft quality

Objective: We hypothesised that the agonal phase prior to cardiac death may negatively influence the quality of the pulmonary graft recovered from non-heart-beating donors (NHBDs). Different modes of death were compared in an experimental model. Methods: Non-heparinised pigs were divided into three groups (n = 6 per group). Animals in group I [FIB] were sacrificed by ventricular fibrillation resulting in immediate circulatory arrest. In group II [EXS], animals were exsanguinated (45 ± 11 min). In group III [HYP], hypoxic cardiac arrest (13 ± 3 min) was induced by disconnecting the animal from the ventilator. Blood samples were taken pre-mortem in HYP and EXS for measurement of catecholamine levels. After 1 h of in situ warm ischaemia, unflushed lungs were explanted and stored for 3 h (4 °C). Left lung performance was then tested during 60 min in our ex vivo reperfusion model. Total protein concentration in bronchial lavage fluid was measured at the end of reperfusion. Results: Pre-mortem noradrenalin (mcg l⁻¹) concentration (baseline: 0.03 ± 0) increased to a higher level in HYP (50 ± 8) vs EXS (15 ± 3); p = 0.0074. PO₂ (mmHg) at 60 min of reperfusion was significantly worse in HYP compared to FIB (445 ± 64 vs 621 ± 25; p < 0.05), but not to EXS (563 ± 51). Pulmonary vascular resistance (dynes s cm⁻⁵) was initially higher in EXS (p < 0.001) and HYP (NS) vs FIB (15824 ± 5052 and 8557 ± 4933 vs 1482 ± 61, respectively) but normalised thereafter. Wet-to-dry weight ratio was higher in HYP compared to FIB (5.2 ± 0.3 vs 4.7 ± 0.2, p = 0.041), but not to EXS (4.9 ± 0.2). Total protein (g l⁻¹) concentration was higher, although not significant in HYP and EXS vs FIB (18 ± 6 and 13 ± 4 vs 4.5 ± 1.3, respectively). Conclusion: Pre-mortem agonal phase in the NHBD induces a sympathetic storm leading to capillary leak with pulmonary oedema and reduced oxygenation upon reperfusion. Graft quality appears inferior in NHBD lungs when recovered in controlled (HYP) vs uncontrolled (EXS and FIB) setting.     

依布西林对大鼠无心跳供体肺的保护作用

目的 观察依布西林在大鼠无心跳供体(NHBD)肺保护中的作用.方法 将60只SD大白鼠随机分为A组:有心跳供体(HBD)组;B组:NHBD组;C组:NHBD+依布硒林(Ebselen)组.B组、C组供体处死后维持辅助呼吸,放置室温中30 min,再灌注低钾右旋糖苷(LPD)液.受体鼠行"原位左肺移植术".C组受体在肺移植前1 h给予Ebselen.结果 C组移植后肺顺应性为0.1740±0.0100,结扎右肺门后15、30 min动脉血氧分压分别为(93.97±5.94)、(92.30±6.57)mm-Hg,肺组织丙二醛(MDA)含量为(0.63±0.23)nmol/mg蛋白,肺组织能量代谢物总量为(821.51±29.70)mol/g,与B组比较差异均有统计学意义(P＜0.05).结论 给予受体一定浓度的Ebselen可改善NHBD肺保护作用.

Abstract：

Objective To evaluate the protective effect of ebselen on the rat lungs from non-heartbeating donors (NHBD). Methods Sixty Sprague-Dawley rats were randomly divided into 3 groups:group A, heart-beating donor; group B, NHBD with 30 min of warm ischemia time (WIT); group C, NHBD with 30 min of WIT and administration of ebselen. The donor lungs in groups B and C maintained ventilation at room temperature for 30 min after asystolia and then were flushed with LPD solution. The recipient rats underwent left lung transplantation. The recipients in group C were administered with ebselen 1 h before transplantation. Results All the recipients survived during the observation period. The pulmonary compliance of group C was 0. 1740 ±0. 0100. The PaO2 at 15 min and 30 min after the ligation of the right pulmonary hila was (93.97 ±5.94), (92. 30 ±6. 57) mmHg, respectively. Malondialdehyde (MDA) of the pulmonary tissue was (0. 63 ±0. 23) nmol/mg pro and the energy metabolism was (821.51 ±29.70)mol/g. The difference between group B and group C was significant (P ＜ 0. 05 ). Conclusion The administration of ebselen is a safe and effective treatment in the preservation of the rat lungs from NHBD.     

Retrograde arterial flush of the liver graft in living donor liver transplantation may ameliorate post-transplantational cholestasis--prospective randomized study

Retrograde arterial flush (RGAF) of liver grafts, which prevents arterial intima injuries in living donor liver transplantation (LDLT), seemed to reduce post-transplantational cholestasis by our preliminary study. The aim of this study was to further confirm this effect of RGAF of grafts in LDLT by a prospective randomized study. The enrolled cases were randomly divided into two groups, RGAF (portal flush plus RGAF) and non-RGAF (portal flush only). The outcome measures included intra-operative hemodynamic changes, one-month post-transplantational liver function tests and acute cellular rejection, vascular and biliary complications, the length of postoperative hospital stay, and graft and patient survivals. The significant findings of results were lower postoperative serum bilirubin in the RGAF group until three wk after transplantation (days 1, 3, 7, 14, and 21 serum total bilirubin, mg/dL: 5.82±5.46, 3.95±4.28, 4.87±5.01, 3.62±5.32, and 3.63±6.91 vs. 8.41±5.49, 7.47±5.61, 10.0±10.1, 9.06±11.6, and 6.02±9.84, respectively, in RGAF and non-RGAF, p=0.017, 0.002, 0.014, 0.011, 0.109) and shorter postoperative hospital stay in the RGAF group (31.1±17.6 vs. 44.8±32.3 d, respectively, in RGAF and non-RGAF, p=0.035). In conclusion, the RGAF of liver grafts on a back table may ameliorate postoperative functional cholestasis in LDLT, which possibly reduces postoperative hospital stay.     

双层法氧合冷保存心跳停搏大鼠肝细胞移植研究

目的 观察双层法(TLM)氧合冷保存较UW保存能否改善心跳停搏供体(NHBD)肝细胞存活率和功能.方法 SD大鼠为供体,建立NHBD模型,NAPs大鼠为受体.根据热缺血时间(WIT)15 m/n和30 m/n分成2组;按TLM、UW分别保存3、12 h和未保存再各分5个亚组(n=5).检测NHBD肝细胞存活率和ATP水平,观察肝细胞移植(HTx)后肝细胞形态和功能.结果 TLM3、12 h组肝细胞存活率分别显著高于UW 3、12 h组[(69.7±4.1)%和(69.1±2.0)%比(55.1±2.3)%和(53.3±2.0)%;P<0.01];TLM 3、12 h组AlP水平分别显著高于UW 3、12 h组(3.25±0.79和3.06±0.67比2.25±0.53和1.63±0.40;P<0.05或P<0.01).HTx后几乎所有时间点TLM组血清白蛋白(ALB)水平都显著高于UW组(P<0.05或P<0.01).在HTx 14d后,形态学显示TLM组肝细胞保持强活力,糖原和ALB染色呈强阳性.结论 TLM氧合冷保存可显著改善和逆转NHBD肝细胞存活率和功能,减少NHBD肝细胞缺血性损伤.     

Effect of machine perfusion preservation on delayed graft function in non-heart-beating donor kidneys – early results

The functioning of non-heart-beating (NHB) donor kidneys upon transplantation is often delayed. To evaluate the effect of preservation by machine perfusion (MP) on early post-transplant function, 37 NHB donor kidneys were compared to 74 matched heart-beating (HB) donor kidneys preserved by cold storage (CS). The NHB donor kidneys were subject to 49 ± 34 min of warm ischemia. Delayed function (DF) and primary nonfunction (PNF) rates were significantly higher for NHB than for HB donor kidneys (49 % and 19 % vs 34 % and 7 %, respectively). Consequently, renal function was impaired but recovered within 6 months. MP could not eliminate the differences in DF rate between NHB and HB donor kidneys. However, NHB donor kidneys preserved by MP showed less DF than that reported in kidneys preserved by CS. This suggests that MP has a beneficial effect on ischemically damaged kidneys. The similar results observed with category 2 and category 3 NHB donors also suggest this effect. The high PNF rate emphasizes the need for viability tests that prevent the transplantation of nonviable organs. We conclude that MP alone is not sufficient to reduce DF and PNF rates in NHB donor kidneys. Received: 16 January 1997 Received after revision: 7 April 1997 Accepted: 11 April 1997     

Retrograde flush is more protective than heparin in the uncontrolled donation after circulatory death lung donor

Background

Formation of microthrombi after circulatory arrest is a concern for the development of reperfusion injury in lung recipients from donation after circulatory death (DCD) donors. In this isolated lung reperfusion study, we compared the effect of postmortem heparinization with preharvest retrograde pulmonary flush or both.

Methods

Domestic pigs (n = 6/group) were sacrificed by ventricular fibrillation and left at room temperature for 1 h. This was followed by 2.5 h of topical cooling. In control group [C], no heparin and no pulmonary flush were administered. In group [R], lungs were flushed with Perfadex in a retrograde way before explantation. In group [H], heparin (300 IU/kg) was administered 10 min after cardiac arrest followed by closed chest massage for 2 min. In the combined group, animals were heparinized and the lungs were explanted after retrograde flush [HR]. The left lung was assessed for 60 min in an ex vivo reperfusion model.

Results

Pulmonary vascular resistance at 50 and 55 min was significantly lower in [R] and [HR] groups compared with [C] and [H] groups (P < 0.01 and P < 0.001) and at 60 min in [R], [H], and [HR] groups compared with [C] group (P < 0.001). Oxygenation, compliance, and plateau airway pressure were more stable in [R] and [HR] groups. Plateau airway pressure was significantly lower in [R] group compared with the [H] group at 60 min (P < 0.05). No significant differences in wet–dry weight ratio were observed between the groups.

Conclusions

This study suggests that preharvest retrograde flush is more protective than postmortem heparinization to prevent reperfusion injury in lungs recovered from donation after circulatory death donors.     

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。     

Transimission of donor lymphocytes in clinical lung transplantation

Passenger mononuclear cells in organ grafts are known to influence the alloimmune response to the graft. To assess their relevance in clinical lung transplantation, we studied the amount, distribution, cell types, and surface marker expression of mononuclear cells in human donor lungs. Two major compartments of mononuclear cells could be differentiated: lymph nodes containing resting T and B lymphocytes, and the lung tissue itself, containing mainly activated lymphocytes as well as monocytes/macrophages. Tissue-associated mononuclear cells make up 20–40x10⁹ cells per lung, about 30–50 % of which are lymphocytes. Tissue-associated lymphocytes are predominantly T and NK cells; most of the T cells are CD8⁺ CD45R0⁺ and express HLA-DR. Strong expression of the adhesion molecules LFA-1 and ICAM-1 is present on infiltrating cells as well as on resident cells of the organ. Moreover, the lymphocytes inside the lung tissue are functionally highly active, with a strong stimulatory as well as alloreactive potency. Thus, large numbers of allogeneic mononuclear cells and particularly large numbers of functionally active lymphocytes are obviously transmitted by human lung allografts. The immunological in vivo relevance of these cells after lung transplantation may include allostimulation and graft-versus-host activity, but also beneficial immunomodulatory effects.     

Procurement of lungs for transplantation following donation after circulatory death: the Alfred technique

Introduction

Donation after circulatory death (DCD) is an evolving method for lung transplantation (LTx) with results comparable to donation after brain death (DBD).

Materials and Methods

DCD lung transplant program requires a systematic approach for an efficient utilization of hospital resources. The surgical techniques have been developed to minimize the ischemic time during lung procurement. We have presented our management protocol and the surgical techniques as used at the Alfred Hospital in Melbourne, Australia.

Results

We have transplanted 92 recipients with lungs procured from 91 donors over an 8 year period from May 2006 to July 2014. This accounted for an extra 19% lung transplant operations performed during this time period. Operative mortality was 1% and 8 year survival was 71% in DCD lung recipients.

Conclusions

DCD lung transplantation provides an additional significant pool of lung donors with satisfactory short and long term outcomes.     

Antegrade versus retrograde perfusion of the donor lung: impact on the early reperfusion phase

Transpulmonary thermodilution was used to evaluate the effect of flush route during harvest on hemodynamic and respiratory function of the pulmonary graft in the early post-transplant phase. Single lung transplantation was performed in two piglet groups after 24 h of cold storage. Donor organs for group A underwent antegrade perfusion, and those for group R retrograde perfusion. PaO₂, compliance (C), airway resistance (R), extravascular lung water index (EVLWI), pulmonary blood volume index (PBVI), intrathoracic blood volume index (ITBVI), capillary leak (CL), and cardiac function index (CFI) were assessed by transpulmonary thermodilution at baseline, 1, 3, and 6 h after reperfusion. EVLWI was significantly lower in group R. Compliance and PaO₂ were higher in the same group. The two groups did not differ significantly with regard to CFI, PBVI, ITBVI, and airway resistance. Retrograde perfusion of the donor lung had a positive impact on graft function during early reperfusion. Transpulmonary hemodynamic monitoring can be a powerful tool for intra- and postoperative management of transplant patients.     

Outcome of machine-perfused non-heart-beating donor kidneys, not allocated within the Eurotransplant area

Abstract Eleven non-heart-beating (NHB) donor kidneys considered vital during machine perfusion (MP), could not be allocated inside Eurotransplant (ET). With the help of ET, five kidneys were transplanted in Karachi and six in Basel. Our goal was to prove that NHB kidneys successfully passing MP viability tests can be transplanted safely. Methods: Donor age, serum creatinine (some post-mortem) and warm ischaemic time were, respectively, (mean and range): 44 (14–70) years, 137 μmol/1, and 44 (9–80) min. Reasons for refusal were: bold ureter (one), suboptimal flush (one), relatively long hypotensive phase (seven), and donor age of 70 years (two). After 8 h of MP, mean lactate dehydrogenase, intrarenal resistance and alpha glutathione S-transferase were (including range): 556 U/l (range 366–819 U/l), 0.86 mmHg/ml per min (0.41-1.15 mmHg/ml per min) and 1188 μg/1 (575–2677 μg/1), respectively. Mean cold ischaemic time was 45 (range 28–72) h. Results: Two kidneys showed immediate function, and nine showed delayed function. Mean creatinine levels after 1, 3 and 6 months were 295, 200 and 206 μmol/1, respectively. Four patients died for reasons not related to their kidney transplantation. Conclusions: We claim that MP can successfully assess viability of NHB donor kidneys. The reluctance to accept MP, and judged vital, NHB donor kidneys is no longer justified.     

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     

机器灌注在心脏死亡捐献供肾保存中的应用进展

机器灌注在保存心脏死亡捐献(DCD)供肾过程中,比单纯静态冷保存具有更多的优越性。随着我国DCD供肾的应用,机器灌注已受到各大移植中心的重视,并越来越多地应用于捐献器官的保存。本文就机器灌注的工作原理、机制及在保存DCD供肾过程中的应用研究进展作一综述。     

乌司他丁减轻无心跳大鼠供肺缺血再灌注损伤的作用及机制

目的 研究乌司他丁(UTI)减轻无心跳大鼠供肺缺血再灌注损伤作用及作用机制.方法 选取雄性SD大鼠作为供、受鼠建立无心跳大鼠左肺移植模型,将受鼠分为对照组和实验组,对照组受鼠接受的供肺经低钾右旋糖酐(LPD)液灌注和保存,实验组受鼠接受的供肺经含乌司他丁(50万U/L)的LPD液灌注和保存.移植过程中监测受鼠的动脉血氧合情况;移植肺再灌注30 min和1h时,取两组受鼠移植肺组织,测量和计算湿干质量比,检测移植肺组织中丙二醛(MDA)和超氧化物歧化酶(SOD)含量;提取RNA,采用实时定量聚合酶链反应检测移植肺组织中肿瘤坏死因子-α(TNF-a)、细胞间黏附分子-1(ICAM-1)和白细胞介素10(IL-10) mRNA的相对表达量.结果 再灌注后1h,实验组的氧合指数(PaO2/FiO2)为472.38±31.66,显著高于对照组的429.52±14.83,两组比较,差异有统计学意义(P=0.025).与对照组相比,实验组在再灌注30 min和1h时的水肿情况(湿干质量比)均好于对照组(P=0.005,P=0.006),实验组移植肺组织病理损伤也明显轻于对照组.不论再灌注30 min还是1h,实验组移植肺组织中MDA含量较对照组显著降低(P=0.039,P=0.006),而SOD含量显著升高(P=0.035,P=0.030).再灌注30 min时,实验组TNF-a的表达较对照组显著下降(P=0.000),再灌注1h时下降不明显(P=0.139);再灌注30 min时,ICAM-1水平较对照组下降不明显(P=0.062),再灌注1h则存在明显降低(P=0.001);再灌注30 min和1h时,实验组IL-10 mRNA的表达水平均较对照组显著上调(P=0.004,P=0.000).结论 乌司他丁能够减轻无心跳大鼠供肺的缺血再灌注损伤,对移植肺有保护作用.     

Retrograde Arterial Flush of the Liver Graft in Living Donor Liver Transplantation

ABSTRACT?

Formal hepatic arterial flush to preserve the liver graft in living donor liver transplantation (LDLT) is not recommended by most transplant centers because direct cannulation may injure the intima of the hepatic artery. The authors describe a method of retrograde arterial flush of the liver graft without arterial cannulation by hepatic venous outflow occlusion (HVOO) in LDLT. First, we proved no backflow of the hepatic artery without HVOO by portal flush to pig livers. Then we used HVOO on 15 LDLT cases (Group HVOO). The results were compared with those of 24 counterpart LDLT cases (Group non-HVOO) without hepatic artery flush. The two-week posttransplantational liver functions were not different between two groups except that the day-three and day-seven serum bilirubin levels were lower in Group HVOO (day-three total bilirubin: 4.99 ± 4.04 mg/dl versus 7.65 ± 4.33 mg/dl, p =.016; day-seven total bilirubin: 5.06 ± 5.02 mg/dl versus 9.57 ± 6.09, p =.005). The rates of vascular complications, six-month graft survival, and biliary anastomotic stricture in Group HVOO were 0, 93.3, and 13.3% respectively, which were not different from those of Group non-HVOO. In summary, to avoid intima injury, the retrograde arterial flush of liver graft by HVOO is safe in LDLT. The short-term results showed the effect of decreasing early functional cholestasis but the long-term benefits need further evaluation, especially with regards to biliary anastomotic complications.     

Expanding the donor pool: use of renal transplants from non-heart-beating donors supported with extracorporeal membrane oxygenation

Abstract:  In response to organ shortage, we used the renal grafts from non-heart-beating donors (NHBDs). Extracorporeal membrane oxygenation (ECMO) was used to maintain NHBDs before organ procurement. We compared the results of renal transplantation from different donors, including heart-beating donors (HBDs), living-related donors (LDs), and NHBDs supported with ECMO. From February 1998 to June 2003, we recruited 219 patients receiving renal transplantation at National Taiwan University Hospital. Among them, 31 received kidneys from NHBDs supported with ECMO, 120 from HBDs, and 68 from LDs. Multiple organ transplant recipients were not included in this study. We compared the graft survival, serum creatinine levels, and estimated glomerular filtration rates of the three groups. The rate of delayed graft function was higher in NHBD recipients (41.9%) than in HBD recipients (27.0%) and LD recipients (10.9%) (p = 0.003). In the NHBD group, the recipients of grafts with delayed function had significantly longer ECMO runs (63.1 ± 3.0 min) than those without delayed function (53.7 ± 2.5 min) (p = 0.024). Estimated glomerular filtration rate (p = 0.472) and mean serum creatinine level (p = 0.286) were not significantly different between the three groups using a longitudinal approach. The 5-yr graft survival rates for NHBD (88.4%, 95% CI: 0.680–0.962), HBD (83.2%, 95% CI: 0.728–0.899), and LD transplant recipients (89.3%, 95% CI: 0.619–0.974) were not significantly different (p = 0.239). The 5-yr patient survival rates for NHBD, HBD, and LD transplant recipients were 100, 93.0 (95% CI: 0.859–0.966) and 100% respectively. The long-term allograft survival and function of kidneys from NHBDs supported by ECMO, HBD, and LD did not differ significantly. Long ECMO running time tended to delay graft function.     

不同受者同期分别接受同一供者的心、肺移植六例

目的为提高供者器官的利用率，探讨将同一供者的心、肺同期移植给不同受者的可行性及手术方法。方法采用原位灌洗、整块切取的方法获得3例供者的心脏和双肺，分别同期移植给3例终末期心脏疾病患者和3例终末期肺脏疾病患者。供者先行机械通气，然后开胸，主动脉及肺动脉分别插入灌注管，经主动脉插管灌注HTK液，经肺动脉插管灌注改良棉子糖低钾右旋糖酐液，对心、肺进行灌洗，然后整块切取心、肺。受者手术前进行供心、肺的分离，然后再次进行心、肺的逆行灌洗（通过上、下肺静脉插管和主动脉根部或者经腔静脉冠状窦逆行插管）。施行单侧肺移植者，再将左、右供肺分离。3例肺移植受者中，1例行左单肺移植，其他2例为非体外循环下序贯式双肺移植；3例心脏移植者均采用双腔静脉吻合法施行原位心脏移植。结果1例左单肺移植受者术后恢复良好；1例双肺移植受者术后出现右肺上叶静脉栓塞，于术后第9天再次手术，切除右肺上叶，后痊愈出院；另1例双肺移植受者，术后出现支气管吻合口狭窄，并发肺部感染，经对症治疗后好转出院，术后7个月死于重症感染。1例心脏移植受者术后早期出现肾功能衰竭，经血液透析治疗后痊愈出院，3例心脏移植受者术后生活质量良好，心功能恢复到0-I级。结论将同一供者的心、肺分别移植给不同受者，能充分利用供者器官，方法可行，效果良好。