Development of a cost-effective ex vivo lung perfusion system for lung transplantation in India

Background & objectives: Standard donor lung preservation with cold flush and storage allows up to six hours between retrieval of lungs from the donor and transplantation in the recipient. Ex vivo lung perfusion (EVLP) systems mimic physiological ventilation and perfusion in the donor lungs with potential for prolonged lung preservation and donor lung reconditioning. In this study, it was aimed to perform EVLP on discarded donor lungs using a locally developed EVLP system. Methods: Equipment that are routinely used for cardiac surgeries were collected and a functional EVLP system was assembled. This system was used on five pairs of lungs retrieved from brain-dead organ donors. The lungs were ventilated and pulmonary circulation was continuously perfused with a solution containing oxygen and nutrients for four hours. The system was tested without red blood cells (RBCs) added to the solution (acellular group; n=3; A1, A2 and A3) and also with RBCs added to the solution (cellular group; n=2; C1 and C2). Results: The EVLP system was successfully used in four (A1, A2, A3 and C2) of the five lung pairs. Mechanical and gas exchange functions of the lungs were preserved in these lung pairs. One lung pair (C1) worsened and developed pulmonary oedema. Histopathological examination of all five lung pairs was satisfactory at the end of the procedure. Major challenges faced were leakage of solution from the system and obstruction to drainage of RBCs containing solution from the lungs. Interpretation & conclusions: The results of the present study suggest that, it is possible to maintain the lungs retrieved for transplantation in a physiological condition using a locally prepared EVLP system and a solution without RBCs.

Preservation fluid cultures. Clinical significance in liver transplantation / Cultivos de líquido de preservación en trasplante hepático. Significancia clínica

Abstract The aim of this study was to determine the incidence of preservation fluids (PF) bacterial positive cultures, identify the germs involved, determine their correlation with infections in recipients during the postoperative period and compare outcomes in terms of morbidity, hospital stay and both patient and graft survival. We describe incidence and etiology of germs developed in PF cultures in our series and evaluate its impact on recipients. A prospective study in deceased donor liver transplants (LT) recipients was carried out from January 2014 to December 2017. Back table PF cultures were analized considering positive the development of any germs and negative to no signs of growth after 5 days. PF were classified as contamination or pathogens. Targeted antibiotic therapy was administered in the last ones. Recipients were divided in: PF (-) and PF(+). Recipients infections related to positive PF were analyzed. These were identified as "direct correlation" when the same germ grew up in PF. Hospital stay and 30 days follow up were compared. Eighty-eight patients PFs were included, 38% (33) had positive cultures, 28 (85%) of these were considered contamination and only 5 as pathogens. We found no differences in postoperative infections (p 0.840), ICU and total hospital stay (p 0.374 and 0.427) between both groups. Postoperative infections and hospital stay seem not to be influenced by PF cultures positivity. Treatment of isolated pathogens could have prevented infections, therefore, those groups that perform PF cultures should consider treatment in these cases and conclude prophylaxis when PF is negative or contaminated.

Resumen Las infecciones bacterianas son frecuentes en pacientes sometidos a trasplante hepático. Describimos la incidencia y etiología de los cultivos de líquidos de preservación (LP) positivos en nuestra serie y analizamos su importancia clínica. Se trata de un trabajo prospectivo de pacientes trasplantados hepáticos, entre enero 2014 a diciembre 2017. Se analizaron muestras de LP tomadas al finalizar la mesa de banco, considerándose positivo el desarrollo de cualquier germen y negativo la ausencia del mismo luego de 5 días. Los LP positivos se clasificaron en: con contaminantes y con patógenos. Los pacientes con LP patógenos recibieron tratamiento antibiótico de acuerdo al antibiograma. Los pacientes fueron divididos en dos grupos: con LP + y LP-. Las infecciones relacionadas a los LP fueron analizadas. Se consideró "correlación directa" cuando el mismo germen desarrolló en el LP y en el recipiente. Se comparó estadía hospitalaria en ambos grupos. Se incluyeron 88 pacientes, 38% (33) presentaron LP+, de los que el 85% (28) fueron por contaminación y 5 por pa tógenos. No se hallaron diferencias significativas en infecciones postoperatorias (p 0.840) y estadía hospitalaria (p 0.427) entre ellos. No hubo casos de "correlación directa". Las infecciones postoperatorias y la estadía hospitalaria de los pacientes no parecen estar influidas por la positividad de los cultivos de LP. El tratamiento dirigido a los gérmenes aislados como patógenos pudo prevenir infecciones, por lo tanto, los grupos que realizan cultivos de rutina deberían considerar el tratamiento en estos casos y finalizar la profilaxis cuando el LP sea negativo o contaminado.

Original and generic preservation solutions in organ transplantation. A new paradigm?

Abstract Solid organ transplantation is a very complex process, in which the storage of the graft in a preservation solution is mandatory in order to extend ischemic times and contain further damage. The condition in which the organ is transplanted is critical for the outcome of the organ recipient. The recent emergence of generic versions of organ preservation solutions (solutions with the same composition and under the same legislation as the original versions, but with different brands) compelled us to study whether the standards are maintained when comparing the original and its generic counterpart. Along these lines, we discuss and comment on some aspects concerning this issue of general interest in the organ transplantation field.

Effect of a new organ preservation solution with HOE642 on the apoptosis of the donor lung from a modified lung transplantation model of rabbits / 重庆医学

Objective To explore the effect of a new organ preservation solution with HOE642 on the apoptosis of the donor lung from a modified lung transplantation model of rabbits.Methods 24 male rabbits were divided into two groups [low potassium dextran (LPD) group and HOE group],established rabbit models for next 2-h reperfusion using LPD solution or new organ preservation solution.Detected the levles of apoptosis index and caspase-3,the expression of Fas/Fas-L and Bcl-2/Bax.Results Compared with LPD group,HOE group revealed significant lower level of apoptosis index and caspase-3 (P＜0.05),lower expression of Fas/Fas-L and Bax,and higher expression of Bcl-2 (P＜0.05).Conclusion The potential donor lung protective mechanism offered by the new organ preservation solution with HOE642 might be the inhibition of apoptosis via both intrinsic and extrinsic pathways.

Alternative solution for ex vivo lung perfusion, experimental study on donated human lungs non-accepted for transplantation

PURPOSE: To evaluate a new perfusate solution to be used for ex vivo lung perfusion. METHODS: Randomized experimental study using lungs from rejected brain-dead donors harvested and submitted to 1 hour of ex vivo lung perfusion (EVLP) using mainstream solution or the alternative. RESULTS: From 16 lungs blocs tested, we found no difference on weight after EVLP: Steen group (SG) = 1,097±526g; Alternative Perfusion Solution (APS) = 743±248g, p=0.163. Edema formation, assessed by Wet/dry weigh ratio, was statistically higher on the Alternative Perfusion Solution group (APS = 3.63 ± 1.26; SG = 2.06 ± 0.28; p = 0.009). No difference on PaO2 after EVLP (SG = 498±37.53mmHg; APS = 521±55.43mmHg, p=0.348, nor on histological analyses: pulmonary injury score: SG = 4.38±1.51; APS = 4.50±1.77, p=0.881; apoptotic cells count after perfusion: SG = 2.4 ± 2.0 cells/mm2; APS = 4.8 ± 6.9 cells/mm2; p = 0.361). CONCLUSION: The ex vivo lung perfusion using the alternative perfusion solution showed no functional or histological differences, except for a higher edema formation, from the EVLP using Steen Solution(r) on lungs from rejected brain-dead donors. .

Preliminary study of coconut water for graft tissues preservation in transplantation / Estudo preliminar da água de coco para preservação de enxertos teciduais em transplante

OBJECTIVE: To verify the effectiveness of coconut water in preserving tissues for transplant. METHODS: Fifty male Wistar rats were randomly distributed in five groups, according to the following preservation solutions for tissue grafts: Group 1: Lactated Ringer; Group 2: Belzer solution; Group 3: mature coconut water; Group 4: green coconut water; Group 5: modified coconut water. In Group 5, the green coconut water has been modified like the Belzer solution. From each animal we harvasted the spleen, ovaries and skin of the back segment. These tissues were preserved for six hours in one of the solutions. Then, the grafts were reimplanted. The recovery of the function of the implanted tissues was assessed 90 days after surgery, by splenic scintigraphy and blood exame. The implanted tissues were collected for histopathological examination. RESULTS: The serum levels did not differ among groups, except for the animals in Group 5, which showed higher levels of IgG than Group 1, and differences in relation to FSH between groups 1 and 2 (p <0.001), 4 and 2 (p = 0.03) and 5 and 2 (p = 0.01). The splenic scintigraphy was not different between groups. The ovarian tissue was better preserved in mature coconut water (p <0.007). CONCLUSION: the coconut water-based solutions preserves spleen, ovary, and rat skin for six hours, maintaining their normal function.

OBJETIVO: Verificar a eficácia da água de coco na preservação de tecidos para transplante. MÉTODOS: cinquenta ratas Wistar foram distribuídas aleatoriamente em cinco grupos, de acordo com as seguintes soluções de preservação para enxertos teciduais: Grupo 1- Ringer lactato, Grupo 2- Solução de Belzer, Grupo 3- Água de coco maduro, Grupo 4- Água de coco verde, Grupo 5- Água de coco modificada. No Grupo 5, a água de coco verde foi modificada à semelhança da solução de Belzer. De cada animal, retirou-se o baço, os ovários e um segmento de pele do dorso. Esses tecidos foram preservados durante seis horas em uma das soluções. Em seguida, os enxertos foram reimplantados. A recuperação da função dos tecidos implantados foi avaliada 90 dias após a cirurgia, por meio de cintilografia esplênica, exames de sangue. Os tecidos implantados foram coletados para estudo anatomopatológico. RESULTADOS: as dosagens séricas não apresentaram diferença entre os cinco grupos, exceto pelos animais do Grupo 5, que apresentaram valores mais elevados de IgG do que o Grupo 1,e pelas diferenças em relação ao FSH entre os grupos 1 e 2 (p<0,001), 4 e 2 (p=0,03), 5 e 2 (p=0,01). A cintilografia esplênica não foi diferente entre os grupos. O tecido ovariano foi melhor preservado em água de coco maduro (p<0,007). CONCLUSÃO: as soluções à base de água de coco preservam baço, ovário e pele de rato durante seis horas, mantendo sua função normal.

¿Influye el medio de preservación del injerto en los resultados y costos del trasplante hepático de donante fallecido?: Comparación de dos soluciones de preservación / Outcomes using two preservation solutions (UW/HTK) in liver transplantation from brain death donors

Background: Preservation solutions are critical for organ transplantation. In liver transplant (LT), the solution developed by the University Of Wisconsin (UW) is the gold-standard to perfuse deceased brain death donor (DBD) grafts. Histidine-Tryptophan-Ketoglutarate (HTK), formerly a cardioplegic infusion, has been also used in solid organ transplantation. Aim: To compare the outcomes of LT in our center using either HTK or UW solution. Patients and Methods: Retrospective study including 93 LT DBD liver grafts in 89 patients transplanted between March 1994 and July 2010. Forty-eight grafts were preserved with UW and 45 with HTK. Donor and recipient demographics, total infused volume, cold ischemia time, post-reperfusion biopsy, liver function tests, incidence of biliary complications, acute rejection and 12-month graft and patient survival were assessed. Preservation solution costs per liver graft were also recorded. Results: Donor and recipient demographics were similar. When comparing UW and HTK, no differences were observed in cold ischemia time (9.6 ± 3 and 8.7 ± 2 h respectively, p = 0.23), biliary complications, the incidence of acute rejection, primary or delayed graft dysfunction. Histology on post-reperfusion biopsies revealed no differences between groups. The infused volume was significantly higher with HTK than with UW (9 (5-16) and 6 (3-11) l, p < 0.001). The cost per procurement was remarkably lower using HTK. Conclusions: Perfusion of DBD liver grafts with HTK is clinically equivalent to UW, with a significant cost reduction.

Comparative efficacy of Belzer or Euro-Collins solutions for pancreatic preservation during cold ischemic storage in rats

To compare the efficacy of different types of solutions (Belzer or Euro-Collins) for the preservation of rat pancreas during cold ischemia. METHODS: Thirty Wistar rats were divided into three groups according to the perfusion or storage solution: Group E (perfusion and storage in Euro-Collins solution); Group B (perfusion and storage in Belzer solution) and Group BE (Perfusion in Belzer solution and storage in Euro-Collins solution). After perfusion, the pancreas was excised and stored at 4˚C for 18 hours. Amylase was measured at 6, 12 and 18h, and histological analysis of the pancreas was performed after 18h of cold storage. RESULTS: Amylase was elevated and comparable in Groups E and BE after 12 and 18 hours of ischemia (p<0.05). In the exocrine pancreas, histological differences in the amount of necrosis (p=0.049), lymphocytic infiltrate (p<0.001) and neutrophilic infiltrate (p=0.004) were observed, with more favorable features present in Group B. In the endocrine pancreas, Group B showed less edema (p<0.001), but other parameters were similar among all groups. CONCLUSION: The Euro-Collins solution is inferior to the Belzer solution for the preservation of rat pancreas during cold ischemia.

The impact of topically applied preservation solutions on the respiratory epithelium of tracheal grafts submitted to cold ischemia: functional and morphological analysis

OBJECTIVE: Advances in graft reepithelialization and revascularization have renewed interest in airway transplantation. This study aims to determine whether topically applied preservation solutions can ameliorate ischemic injury to tracheal grafts. We analyzed 1) the effects of cold ischemia on the mucociliary clearance of tracheal grafts and 2) the impact of topically applied preservation solutions on the effects of cold ischemia on mucociliary clearance. METHOD: Tracheal segments (n=217) from 109 male Wistar rats were harvested, submerged in low-potassium-dextran-glucose, histidine-tryptophan-ketoglutarate, or saline solution (saline group), and stored at 4°C for 6, 10, 16, or 24 hours. A control group (not submerged) was analyzed immediately after harvesting. In situ mucociliary transport and ciliary beating frequency were measured using a stroboscope. Epithelial integrity, cellular infiltration, and mucus storage were quantified by light microscopy and image analysis software, along with transmission electron microscopy. RESULTS: 1) The effects of cold ischemia: in situ mucociliary transport and ciliary beating frequency were greater in the control group than after cold ischemia. Microscopic analysis results were similar between groups. 2) The effects of preservation solutions: there was no difference between the low-potassium-dextran-glucose, histidine-tryptophan-ketoglutarate, and saline groups in functional or light microscopy analysis. The saline group presented stronger signs of ischemic injury with transmission electron microscopy. CONCLUSIONS: Cold ischemia diminished the mucociliary clearance of the tracheal respiratory epithelium. Topically applied preservation solutions did not ameliorate the injury caused by cold ischemia to the tracheal respiratory epithelium. .

Comparison of Celsior and Perfadex lung preservation solutions in rat lungs subjected to 6 and 12 hours of ischemia using an ex-vivo lung perfusion system

OBJECTIVE: This study evaluated the performance of lungs that were preserved with different solutions (Celsior, Perfadex or saline) in an ex vivo rat lung perfusion system. METHODS: Sixty Wistar rats were anesthetized, anticoagulated and randomized into three groups (n = 20). The rats were subjected to antegrade perfusion via the pulmonary artery with Perfadex, Celsior, or saline, followed by 6 or 12 hours of ischemia (4ºC, n = 10 in each group). Respiratory mechanics, gas exchange and hemodynamics were measured at 10-minute intervals during the reperfusion of heart-lung blocks in an ex vivo system (IL2-Isolated Perfused Rat or Guinea Pig Lung System, Harvard Apparatus, Holliston, Massachusetts, USA; Hugo Sachs Elektronik, Germany) for 60 minutes. The lungs were prepared for histopathology and evaluated for edema following reperfusion. Group comparisons were performed using ANOVA and the Kruskal-Wallis test with a 5% level of significance. RESULTS: Gas exchange was not significantly different between lungs perfused with either Perfadex or Celsior at the same ischemic times, but it was very low in lungs that were preserved with saline. Airway resistance was greater in the lungs that were preserved for 12 hours. Celsior lungs that were preserved for 6 and 12 hours exhibited lower airway resistance (p = 0.01) compared to Perfadex lungs. Pulmonary artery pressure was not different between the groups, and no significant differences in histopathology and apoptosis were observed between the groups. CONCLUSIONS: Lungs that were preserved with Celsior or Perfadex exhibited similar gas exchange and histopathological findings. Airway resistance was slightly lower in the Celsior-preserved lungs compared with the Perfadex-preserved lungs.

Modelo experimental de perfusão pulmonar ex vivo em ratos: avaliação histopatológica e de apoptose celular em pulmões preservados com solução de baixo potássio dextrana vs. solução histidina-triptofano-cetoglutarato / An experimental rat model of ex vivo lung perfusion for the assessment of lungs regarding histopathological findings and apoptosis: low-potassium dextran vs. histidine-tryptophan-ketoglutarate

OBJETIVO: Comparar os achados histopatológicos e de apoptose em pulmões de ratos preservados em soluções low-potassium dextran (LPD, baixo potássio dextrana), histidine-tryptophan-ketoglutarate (HTK, histidina-triptofano-cetoglutarato) ou salina normal (SN) em 6 h e 12 h de isquemia pela utilização de um modelo experimental de perfusão pulmonar ex vivo. MÉTODOS: Sessenta ratos Wistar foram anestesiados, randomizados e submetidos à perfusão anterógrada pela artéria pulmonar com uma das soluções preservadoras. Após a extração, os blocos cardiopulmonares foram preservados por 6 ou 12 h a 4ºC, sendo então reperfundidos com sangue homólogo em um sistema de perfusão ex vivo durante 60 min. Ao final da reperfusão, fragmentos do lobo médio foram extraídos e processados para histopatologia, sendo avaliados os seguintes parâmetros: congestão, edema alveolar, hemorragia alveolar, hemorragia, infiltrado inflamatório e infiltrado intersticial. O grau de apoptose foi avaliado pelo método TdT-mediated dUTP nick end labeling. RESULTADOS: A histopatologia demonstrou que todos os pulmões preservados com SN apresentaram edema alveolar após 12 h de isquemia. Não houve diferenças em relação ao grau de apoptose nos grupos estudados. CONCLUSÕES: No presente estudo, os achados histopatológicos e de apoptose foram semelhantes com o uso das soluções LPD e HTK, enquanto a presença de edema foi significativamente maior com o uso de SN.

OBJECTIVE: To compare histopathological findings and the degree of apoptosis among rat lungs preserved with low-potassium dextran (LPD) solution, histidine-tryptophan-ketoglutarate (HTK) solution, or normal saline (NS) at two ischemia periods (6 h and 12 h) using an experimental rat model of ex vivo lung perfusion. METHODS: Sixty Wistar rats were anesthetized, randomized, and submitted to antegrade perfusion via pulmonary artery with one of the preservation solutions. Following en bloc extraction, the heart-lung blocks were preserved for 6 h or 12 h at 4ºC and then reperfused with homologous blood for 60 min in an ex vivo lung perfusion system. At the end of the reperfusion, fragments of the middle lobe were extracted and processed for histopathological examination. The parameters evaluated were congestion, alveolar edema, alveolar hemorrhage, inflammatory infiltrate, and interstitial infiltrate. The degree of apoptosis was assessed using the TdT-mediated dUTP nick end labeling method. RESULTS: The histopathological examination showed that all of the lungs preserved with NS presented alveolar edema after 12 h of ischemia. There were no statistically significant differences among the groups in terms of the degree of apoptosis. CONCLUSIONS: In this study, the histopathological and apoptosis findings were similar with the use of either LPD or HTK solutions, whereas the occurrence of edema was significantly more common with the use of NS.

Effects of hydrogen-rich kidney preservation solution on renal cold ischemia/reperfusion injury in rats / 中华麻醉学杂志

Objective To investigate the effects of hydrogen-rich HC-A solution,the self-made kidney preservation solution,on renal cold ischemia/reperfusion (I/R) injury in rats.Methods Twenty-four healthy male Wistar rats,aged 8-10 weeks,weighing 200-250 g,were randomly divided into 3 groups (n =8 each):control group (H1 group),common kidney preservation solution group (H2 group) and hydrogen-rich HC-A kidney preservation solution group (H3 group).In H1 group,only the right kidney was removed.In H2 group,the left kidney was perfused with and cold stored in 4 ℃ common HC-A kidney preservation solution.In H3 group,the left kidney was perfused with and cold stored in a container filled with 4 ℃ common HC-A kidney preservation solution.Blood samples were obtained from the inferior vena cava at 24 h of reperfusion to detect the levels of serum blood urea nitrogen (BUN),creatinine (Cr),TNF-α and IL-6.Left kidneys were removed for determination of malondialdehyde (MDA) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) contents and for examination of the pathological changes in renal tissues (by light microscopy).Results The levels of serum BUN,Cr,TNF-α and IL-6 and contents of MDA and 8-OHdG were significantly higher in H2 and H3 groups than in H1 group,and lower in H3 group than in H2 group (P ＜ 0.05).There were no significant pathological changes in renal tissues in H1 group,the damage to renal tubules was obvious in H2 group and the damage to renal tubules was significantly ameliorated in H3 group as compared with H2 group.Conclusion Hydrogen-rich kidney preservation solution can attenuate renal cold I/R injury in rats.

A multicenter randomized controlled trial of HC-A Ⅱ solution in kidney preservation / 中华器官移植杂志

ObjectiveTo evaluate the efficacy and safety of HC-A Ⅱsolution in kidney preservation.Methods A multicenter,randomized,double-blind and controlled clinical trial was conducted.Between Jan.2008 and Dec.2010,kidney recipients from 9 transplant centers were randomly divided into two groups.Grafts in each group were perfused and stored in HC-A Ⅱ or HTK solutions respectively.Results277 patients were included in the Full Analysis Set (FAS),137 of whom were inHC-A Ⅱgroup and 140inHTK group. Demographic andbaseline medical characteristics were similar between the two groups.262 patients were included in the Per Protocol Set (PPS),133 of whom were in HC-A Ⅱ group and129 in HTK group.The percentages of patients with a serum creatinine level that returned to normal within 28 days postoperation were 86.9％ in HC-A Ⅱ group and 85.0％ in HTK group respectively (P＞0.05 ).The results from PPS analysis were consistent with those from FAS analysis The incidence of test-related adverse events was 2.9％ in HC-AⅡ group and 0.7％ in HTK group respectively (P＞0.05).No test-related serious adverse events occurred throughout the study.ConclusionHC-A Ⅱ solution,the same as HTK solution,is safe and effective in kidney preservation.

Raffinose-low potassium dextran solution in clinical lung transplantation: a retrospective study / 中华器官移植杂志

ObjectiveTo evaluate the security and effectiveness of raffinose-low potassium dextran solution (RLPDs) for donor lung preservation.MethodsFrom Sep.2002 to Dec.2011,131patients underwent lung transplantation in our institution,including 81cases of single lung transplantation and 50 cases of bilateral lung transplantation.There were 95 males and 36 females.A total of 129 donors were used for organ harvest.Two donors denoted their lungs to four patients receiving single lung transplantation. All the donors were flushed, perfused antegradely and retrogradely,and preserved with hypothermal RLPDs.The repeated reperfusion was necessary when the cold ischemic time was fairly long and the function of donor was poor.During operation,the blood gas analysis,mean pulmonary artery pressure (mPAP) and breathing mechanics were monitored.Postoperatively,all the recipients received re-examination of cardiac ultrasonography,pulmonary function,blood gas analysis and chest imaging.ResultsAll lung transplantations were performed successfully,with a mean cold ischemic time of (193±21) min (range,65-630 min).There was a significant relationship between postoperative pneumonedema and donor lung cold ischernic time (P＜0.05),which was also observed between postoperative pneumonedema and primary graft dysfunction (P＜0.01). The blood gas analysis and oxygenation index were significantly improved postoperatively,with the mPAP and parameters of breathing mechanics being descended to a normal level.There were 24 early deaths in the first month post-transplant,with a mortality of 18.3％ (24/131),including 11cases of sepsis,7 cases of primary graft dysfunction,3 cases of heart failure,1case of bronchial stomal leak,1case of acute rejection,and 1case of pulmonary infarction.The pulmonary function was improved significantly 3 months postoperatively.Conclusion RLPDs is efficient in the preservation of donor lung,with the ability to alleviate ischemic reperfusion injury and improve pulmonary function.

Compare the myocardial protection effect of HTK solution with blood cardioplegic solution during valve replacement surgery / 中国医师杂志

Objective To evaluate the effect of HTK solution on myocardial protection during valve replacement surgery.Methods 42 patients with rheumatic heart disease were randomized to receive 4∶1cold blood (control group,n =21 ) and HTK ( protective gronp,n =21 ) cardioplegic solution during valve replacement.The changes of CO and CI were collected at different time points including pre-operation,postoperative 6 hours,12 hours and 24 hours.Aortic clamping time,the ratio of spontaneous cardiac rhythm recovery and inotropic drugs application were calculated,and mechanical ventilation support time and the incidence of arrhythmia were recorded.Results The measurements of CO and CI showed that there was significant higher level in protective group at postoperative 12 hours and 24 hours [ 12 h:(4.82 ± 0.18 ) L/min vs ( 3.50 ± 0.32 ) L/min,( 3.80 ± 0.48 ) L/( min · m2 ) vs (2.79 ± 0.39) L/( min · m2 ) ;24 h:(4.97±0.45)L/min vs ( 3.81 ±0.19)L/min,(4.22±0.17)L/(min · m2) vs (2.91 ±0.21)L/(min·m2 ),P ＜ 0.05].The clinical parameters including aortic clamping time,incidence of cardiac arrhythmia,inotropic support,duration of mechanical ventilation and length was lower than in control group [ (53.6 ±24.3 ) min vs ( 68.9 ± 26.1 ) min ; ( 1.8 ± 1.3 ) min vs ( 2.3 ± 1.2 ) min ; ( 33 ± 11 ) min vs ( 42 ± 13 ) min ;(10.2±2.1) μg/(kg · min) vs (15.7 ±3.8) μg/(kg · min);(14.6 ±4.8)h vs (20.7 ±5.1)h,P ＜0.05].The auto-beating rate was higher than in control group (90％ vs 67％,P ＜0.05).Conclusions HTK solution is better than classical blood cardioplegia in myocardial protection during valve replacement.

Evaluation of the presence of microorganisms in solid-organ preservation solution

OBJECTIVE: To assess the presence of microorganism contamination in the preservation solution for transplant organs (kidney/pancreas). Method: Between August 2007 and March 2008, 136 samples of preservation solution were studied prior to graft implantation. Variables related to the donor and to the presence of microorganisms in the preservation solution of organs were evaluated, after which the contamination was evaluated in relation to the "recipient culture" variable. Univariate and multivariate statistical analyses were performed. RESULTS: The contamination rate of the preservation solution was 27.9 percent. Coagulase-negative Staphylococcus was the most frequently isolated microorganism. However, highly virulent agents, such as fungi and enterobacteria, were also isolated. In univariate analysis, the variable "donor antibiotic use" was significantly associated to the contamination of the preservation solution. On the other hand, multivariate analysis found statistical significance in "donor antibiotic use" and "donor's infectious complications" variables. CONCLUSIONS: In this study, 27.9 percent of the preservation solutions of transplant organs were contaminated. Infectious diseases and non-use of antibiotics by the donor were significantly related to the presence of microorganisms in organ preservation solutions. Contamination in organ preservation solutions was not associated with infection in the recipient.

Modelo experimental ex vivo com bloco pulmonar dividido / Ex vivo experimental model: split lung block technique

Modelos de recondicionamento pulmonar ex vivo têm sido avaliados desde sua proposição. Quando são utilizados pulmões humanos descartados para transplante, a grande variabilidade entre os casos pode limitar o desenvolvimento de alguns estudos. No intuito de reduzir esse problema, desenvolvemos uma técnica de separação do bloco pulmonar em direito e esquerdo com posterior reconexão, permitindo que um lado sirva de caso e o outro de controle.

Since they were first established, ex vivo models of lung reconditioning have been evaluated extensively. When rejected donor lungs are used, the great variability among the cases can hinder the progress of such studies. In order to avoid this problem, we developed a technique that consists of separating the lung block into right and left blocks and subsequently reconnecting those two blocks. This technique allows us to have one study lung and one control lung.

Effects of ischemic preconditioning associated to different preservation solutions in protecting the intestinal graft / Efeitos do precondicionamento isquêmico associado a diferentes soluções de preservação na proteção do enxerto intestinal

PURPOSE: To evaluate the effects of ischemic preconditioning (IPC) associate with different preservation solutions, in the protecting of gut. METHODS: Four groups of 14 rats underwent laparotomy and collecting 20 cm of ileum, for preservation, at 4ºC, in Belzer (Belz), Ringer (RL), Celsior (Cs) and Custodiol (Cust) solutions, for 24 hours. Prior to collection, half of the animals in each group were subjected to IPC. During preservation, in the periods of zero, 12, 18 and 24 hours, were conducted evaluating the degree of mucosal injury and dosage of malondialdehyde acid (MDA). RESULTS: In all periods the RL group, with and without IPC, presented MDA values higher than the Belz and Cs. The degree of mucosal injury in the non-ipc RLgroup with 12h preservation was higher than the others; with 18 and 24h, the RL and Cust had higher degrees of damage than Cs and Belz. With IPC, in all periods, the group Cs and Belz had lower degrees of injury. CONCLUSION: The Celsior and Belzer solutions had better protective effects on the gut and these effects were enhanced by IPC.

OBJETIVO: Avaliar os efeitos do precondicionamento isquêmico (PCI) associado a diferentes soluções de preservação, na proteção do intestino delgado. MÉTODOS: Quatro grupos de 14 ratos Wistar, foram submetidos à laparotomia e coleta de 20 cm de íleo, para preservação, a 4ºC, nas soluções de Belzer (Belz), Ringer (RL), Celsior (Cs) e Custodiol (Cust) por 24 horas. Previamente à coleta, em metade dos animais de cada grupo, o intestino foi submetido ao PCI. Durante a preservação, nos períodos de Zero, 12, 18 e 24 horas, foram realizados avaliação do grau de lesão da mucosa e dosagem do ácido malondialdeído (MDA). RESULTADOS: Em todos os períodos o grupo RL, sem e com pci, apresentou valores maiores de MDA do que o Belz e Cs. O grau de lesão da mucosa nos grupos sem pci com preservação de 12h, no grupo RL, foi maior que nos demais; com 18h e 24h o grupo RL e Cust apresentaram maiores graus de lesão do que Cs e Belz. Com o pci, em todos os períodos, os grupos Belz e Cs apresentaram menores graus de lesão CONCLUSÃO: As Soluções Celsior e Belzer tiveram melhores efeitos na proteção do intestino e estes efeitos foram incrementados pelo precondicionamento isquêmico.

Shanghai multi-organ solution in preservation of rat liver: An experimental study / 第二军医大学学报

Objective: To observe the preservative effect of Shanghai-mutil-organ (SMO) solution on rat liver and to assess the feasibility of SMO solution in preserving isolated human livers. Methods: SD rats were randomly divided into 3 groups according to the preservation solutions: SMO group, UW ( University of Wisconsin Solution) group, and HTK (Histidinetryptophan-ketoglutarate solution) group. The simple cold storage liver model was established with isolated rat liver and the liver samples from each group were preserved with the 3 solutions for 8 h, 16 h, 24 h and 36 h. The energy metabolism was analyzed in all the samples and the morphological changes and hepatocytes apoptosis were observed after different preservation periods. Results: The liver tissue contents of ATP, TAN, and AEC in SMO group were significantly higher than those in HTK group at 16 h, 24 h, and 36 h (P<0.05); the contents in SMO group were similar to those in UW group. Histological examination showed that the tissue damage in SMO group was milder than that in HTK group; the damages in SMO group and UW group were similar except that the swelling of cells in SMO group was severer than that in UW group. Apoptosis index in SMO group was lower than that in HTK group at 24 h and 36 h(P< 0.05); there was no significant difference in apoptosis index between SMO group and UW group. Conclusion: SMO solution has a similar preservative effect on rat liver to that of UW solution, only with more severe cell swelling than the latter. The effect of SMO solution is better than that of HTK solution.

Hydrogen-containing preservation solution relieves oxidative stress and inflammatory damage of rat donor heart / 第二军医大学学报

Objective: To study the effect of hydrogen-containing preservation solution against oxidative stress and inflammatory damage of rat donor heart. Methods: Thirty-two SD rats were evenly randomized into four groups(n = 8) : control group (the hearts were protected by HTK solution), H1 group (hydrogen concentration was about 0.2 mmol/L in the HTK solution), H2 group (hydrogen concentration was about 0.4 mmol/L in the HTK solution) and H3 group (hydrogen concentration was about 0.8 mmol/L in the HTK solution). The rat hearts were harvested in all groups and were mounted on the Langendorff apparatus to estimate baseline hemodynamic values. All the hearts underwent hypothermic (4°C) storage for 6 h in the corresponding cardioprotective solutions. Then, superoxide dismutase (SOD) activities and malondiadehyd (MDA) contents in myocardium tissues were measured after reperfusion by xanthine oxidase method and TBA method, respectively. The levels of 8-hydroxydeoxygunosine, TNF-α, and IL-6 in the myocardium tissues were measured by enzyme-linked immunosorbent assay (ELISA). Results: The MDA levels in H1, H2, and H3 groups were lower than that in the control group 6 h after preservation (P<0.01 or P<0.05), and the SOD activities in H2, H3 groups were higher than that in the control group (P<0.01 or P<0.05). The levels of 8-hydroxydeoxygunosine, TNF-α, and IL-6 in the control group were higher than those in H1, H2, and H3 groups (P<0.01 or P<0.05). Conclusion: Hydrogen-containing preservation solution can relieve the oxidative damage and reduce production of inflammation factors during preservation of rat donor heart.