RESUMEN

This study evaluated the current practices of selecting cold storage preservation solutions in Brazil and their impact on delayed graft function (DGF) incidence and 1-year outcomes in kidney transplant recipients. A retrospective cohort study was conducted, including 3,134 brain-dead deceased donor kidney transplants performed between 2014 and 2015 in 18 Brazilian centers. The most commonly used preservation solution was Euro-collins (EC, 55.4%), followed by Histidine-tryptophan-ketoglutarate (HTK, 30%) and Institut Georges Lopez (IGL-1, 14.6%). The incidence of DGF was 54.4%, with 11.7% of patients requiring dialysis for more than 14 days, indicating prolonged DGF. Upon adjusting for confounding variables, HTK demonstrated a significantly lower risk of DGF than EC (OR 0.7350.82500.926), as did IGL-1 (OR 0.6050.7120.837). Similar protective effects were observed for prolonged DGF when comparing HTK (OR 0.4780.5990.749) and IGL-1 (OR 0.4780.6810.749) against EC. No significant association was found between preservation solutions and 1-year death-censored graft survival. In conclusion, EC was the most frequently used cold storage perfusion solution, demonstrating a higher incidence and duration of DGF compared with HTK and IGL-1, but with no impact on 1-year graft survival.

Asunto(s)

Funcionamiento Retardado del Injerto , Trasplante de Riñón , Soluciones Preservantes de Órganos , Preservación de Órganos , Trasplante de Riñón/métodos , Humanos , Brasil/epidemiología , Femenino , Masculino , Estudios Retrospectivos , Adulto , Persona de Mediana Edad , Preservación de Órganos/métodos , Funcionamiento Retardado del Injerto/epidemiología , Supervivencia de Injerto/efectos de los fármacos

RESUMEN

BACKGROUND: Liver transplantation is used for treating end-stage liver disease, fulminant hepatitis, and oncological malignancies and organ shortage is a major limiting factor worldwide. The use of grafts based on extended donor criteria have become internationally accepted. Oxygenated machine perfusion technologies are the most recent advances in organ transplantation; however, it is only applied after a period of cold ischemia. Due to its high cost, we aimed to use a novel device, OxyFlush®, based on oxygenation of the preservation solution, applied during liver procurement targeting the maintenance of ATP during static cold storage (SCS). METHODS: Twenty patients were randomly assigned to the OxyFlush or control group based on a 1:1 ratio. In the OxyFlush group, the perfusion solution was oxygenated with OxyFlush® device while the control group received a non-oxygenated solution. Liver and the common bile duct (CBD) biopsies were obtained at three different time points. The first was at the beginning of the procedure, the second during organ preparation, and the third after total liver reperfusion. Biopsies were analyzed, and adenosine triphosphate (ATP) levels and histological scores of the liver parenchyma and CBD were assessed. Postoperative laboratory tests were performed. RESULTS: OxyFlush® was able to maintain ATP levels during SCS and improved the damage caused by the lack of oxygen in the CBD. However, OxyFlush® did not affect laboratory test results and histological findings of the parenchyma. CONCLUSION: We present a novel low-cost device that is feasible and could represent a valuable tool in organ preservation during SCS.

Asunto(s)

Trasplante de Hígado , Soluciones Preservantes de Órganos , Preservación de Órganos , Humanos , Trasplante de Hígado/métodos , Masculino , Persona de Mediana Edad , Femenino , Preservación de Órganos/métodos , Perfusión/métodos , Perfusión/instrumentación , Prueba de Estudio Conceptual , Oxígeno , Hígado/cirugía , Adenosina Trifosfato , Anciano , Adulto , Donantes de Tejidos/provisión & distribución , Obtención de Tejidos y Órganos/métodos , Isquemia Fría

RESUMEN

Organ transplantations have an increasing medical relevance. It is becoming a regular procedure with an increase in individuals waiting for organs. The increase in the number of discarded organs is mostly due to the donor bacterial and/or viral infection. In this article, we are demonstrating the feasibility of reduction of the bacterial load in kidney model by using ultraviolet-C as a germicidal agent in circulating liquids. Using Staphylococcus aureus as a bacteria model, we were able to demonstrate that in less than 30 min of liquid circulation and associated to irradiation, the bacterial load of the perfusate Custodiol HTK, histidine-tryptophan-ketoglutarate (solution with 5 log CFU mL-1 ), was fully eliminated. A modeling approach was created to verify the possibility of bacterial load decrease, when an organ (here, a renal experimental model) is present in the circuit, releasing a varied rate of microorganisms over time, while the solution is irradiated. Finally, we use an ex vivo model with swine kidney, circulating in the preservation solution with a Lifeport Kidney Transporter machine, to demonstrate that we can contaminate the organ and then promote the elimination of the microbiological load. The results show the feasibility of the technique.

Asunto(s)

Soluciones Preservantes de Órganos , Preservación de Órganos , Animales , Descontaminación , Glucosa , Riñón , Preservación de Órganos/métodos , Perfusión/métodos , Porcinos

RESUMEN

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 infuenced 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.

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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.

Asunto(s)

Trasplante de Hígado , Soluciones Preservantes de Órganos , Contaminación de Medicamentos , Humanos , Trasplante de Hígado/efectos adversos , Donadores Vivos , Estudios Prospectivos , Estudios Retrospectivos

RESUMEN

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.

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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.

Asunto(s)

Humanos , Trasplante de Hígado/efectos adversos , Soluciones Preservantes de Órganos , Contaminación de Medicamentos , Estudios Prospectivos , Estudios Retrospectivos , Donadores Vivos

RESUMEN

OBJECTIVES: Renal transplant with ABO-incompatible donors expands the donor pool. Earlier studies have focused the use of protocol biopsies in ABOincompatible transplant patients. Our study described outcomes of indication (for cause) renal biopsies and clinical outcomes in patients with ABO-incompatible renal transplant. MATERIALS AND METHODS: This retrospective study included 164 patients from January 2012 to June 2019. Biochemical parameters, serial immunoglobulin G anti-ABO titers, and class I and II donor-specific antibody findings were obtained from hospital records, and renal graft biopsies were reviewed according to the Banff 2017 update. RESULTS: We analyzed the results of 65 biopsies from 54 patients. Biopsy-proven acute antibody-mediated rejection (12.8%) was found to be more prevalent than acute cellular rejection (1.8%). Patients with antibodymediated rejection all had microvascular inflammation (g+ptc score of 2 or more, where g+ptc is the sum of the glomerulitis and peritubular capillaritis scores) and were positive for C4d. Acute tubular injury per se was seen in 10.3% of patients; 65% of these patients had C4d positivity in peritubular capillaries, and only 1 patient developed chronic active antibody-mediated rejection on follow-up. Patient and death-censored graft survival rates were 92% and 98% at 1 year after transplant and 88% and 91% at 3 years, respectively. Patients with an episode of antibody-mediated rejection had lower rates of patient (76.5%) and deathcensored graft survival (84.6%) at 1 year. CONCLUSIONS: The microvascular inflammation score (g+ptc score of 2 or higher) is more reliable than diffuse C4d positivity to determine antibody-mediated rejection in ABO-incompatible transplants because diffuse C4d positivity may also be seen in etiologies unrelated to antibody-mediated rejection. Acute tubular injury with C4d positivity without microvascular injury does not confirm antibody-mediated rejection. We suggest that Banff classification be updated in ABOincompatible transplants to include diagnostic criteria for the diagnosis of antibody-mediated rejection.

Asunto(s)

Soluciones Preservantes de Órganos , Biopsia , Complemento C4b , Femenino , Rechazo de Injerto , Humanos , Inflamación , Masculino , Soluciones Preservantes de Órganos/efectos adversos , Fragmentos de Péptidos , Estudios Retrospectivos , Resultado del Tratamiento

RESUMEN

ABSTRACT Purpose: The aim of this study was to evaluate the physical and chemical characteristics of coconut water and to analyze the use of coconut water solution for the conservation of human corneas. Methods: This was an experimental and controlled study performed at the Eye Bank of the General Hospital of Fortaleza. The coconut water-based solution was prepared at the Goat Seed Technology Laboratory of the Department of Veterinary Medicine of the State University of Ceará. Discarded corneas from the Eye Bank were divided into two groups for sequential experiments: G1, coconut water-based solution (experimental group), and G2, conservative treatment with OPTISOL GS® (control group). The osmolality of corneas in G1 was analyzed sequentially at 275, 300, 325, 345, 365, and 400 mOsm/L. The viability of the corneas was determined by specular microscopy and biomicroscopy on the first, third, and seventh days. Results: Corneas preserved in a solution of 365 and 345 mOsm/L had a transparency of 8 mm until the third day and had diffuse edema in the periphery, central folds, and partial epithelium loss until the seventh day. The 365-mOsm/L solution was associated with the worst results during follow-up. Corneas placed in Optisol-GS retained their original aspects. Conclusions: Coconut water-based preservative partially maintained corneal transparency and epithelial integrity, especially during the first three days of follow-up. The coconut water-based solutions used were not effective for use as preservatives in a human eye bank.(AU)

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RESUMO Objetivos: As características físico-químicas e o baixo custo da água de coco foram fundamentais para o este estudo. Analisar o uso de solução a base de água de coco como meio de conservação de córneas humanas em banco de olhos. Métodos: Estudo experimental e controlado realizado no Banco de Olhos do Hospital Geral de Fortaleza. Utilizou-se solução à base de água de coco preparada no laboratório de Tecnologia de Sêmen de Caprinos do Departamento de Medicina Veterinária da Universidade Estadual do Ceará. Foram usadas córneas de descartes divididas em dois grupos: G1 (Conservante com água de coco) - grupo experimental e G2 (grupo Conservante com OPTISOL GS®) grupo controle, em experimentos sequenciais. A osmolaridade do G1 foi analisada sequencialmente com 275, 300, 325, 345, 365 e 400 mOsm/L. A viabilidade das córneas foram realizadas por microscopia especular e biomicroscopia nos 1º, 3º e 7º dias. Resultados: As córneas em solução de 365 e 345 mOsm/L apresentavam transparência nos 8mm centrais até o 3º dia, com edema em toda periferia, dobras centrais e edema 2+, com perda parcial do epitélio até 7º dia, sendo o de maior osmolaridade com melhor transparência durante o seguimento. Grupo com 275, 300 e 400 mOsm/L, córnea opaca, edema difuso, perda total do epitélio no 3º dia. As córneas em Optisol mantiveram seus aspectos. Conclusões: O conservante à base de água de coco manteve em parte a transparência corneana e a integridade epitelial, especialmente nos primeiros 3 dias de seguimento. A solução conservante com água de coco nas formulações utilizadas não se mostrou eficaz para o uso em banco de olhos humanos.(AU)

Asunto(s)

Humanos , Preservación de Órganos/métodos , Biotecnología/métodos , Soluciones Preservantes de Órganos/química , Alimentos de Coco , Bancos de Ojos/organización & administración

RESUMEN

In vitro and in vivo assays were conducted to investigate the effects of trans-resveratrol (RVT) on liquid-extended boar semen during 72 h of storage at 17 °C. Thirty-six ejaculates were collected from six boars, evaluated, and extended. RVT was then added at the indicated treatment concentration (0, 0.01, 0.1 or 1 mM), and the ejaculates were cooled to 17 °C and evaluated at 0, 24, 48, and 72 h. Samples were evaluated for sperm motility, kinetics, plasma and acrosome integrity, mitochondrial membrane potential, anion superoxide levels, lipoperoxidation, and antioxidant enzyme activity. In the follow-up experiment, twenty-eight gilts were fixed-time inseminated with 0 or 0.01 mM RVT liquid-extended boar semen. After five days, they were slaughtered, and their reproductive tracts were recovered. The embryos were collected, and the pregnancy, fertility, and viable embryo rates were calculated. In the in vitro assays, total motility, plasma and acrosome membrane integrity, mitochondrial membrane potential, anion superoxide levels, and lipoperoxidation did not change at any of the evaluation times with the use of RVT up to 0.01 mM. RVT decreased SOD activity without changes in GPx. RVT used at 1 mM showed harmful effects for almost all evaluated parameters. For the in vivo assay, the same pregnancy and fertility rates were observed for both groups, while the viable embryo rate was three-fold lower in the 0.01 mM group than in the 0 mM group. The results showed a dichotomous effect of RVT; a low concentration was not harmful in vitro but was catastrophic for embryo viability.

Asunto(s)

Fertilidad/efectos de los fármacos , Resveratrol/farmacología , Preservación de Semen/veterinaria , Semen/efectos de los fármacos , Espermatozoides/efectos de los fármacos , Porcinos , Acrosoma/efectos de los fármacos , Animales , Antioxidantes/farmacología , Femenino , Inseminación Artificial/veterinaria , Masculino , Soluciones Preservantes de Órganos/farmacología , Embarazo , Análisis de Semen/veterinaria , Preservación de Semen/métodos , Motilidad Espermática/efectos de los fármacos , Superóxidos

RESUMEN

Purpose To compare Fructose-1,6-Bisphosphate (FBP) to Histidine-Tryptophan-Ketoglutarate (HTK) in liver preservation at cold ischemia. Methods Male rats (Sprague-Dawley: 280-340g) divided into three groups (n=7): Control; Fructose-1,6-bisphosphate (FBP); Histidine-Tryptophan-Ketoglutarate (HTK). Animals underwent laparotomy-thoracotomy for perfusion of livers with saline. Livers were removed and deposited into solutions. Mitochondria were isolated to determine State 3 (S3), State 4 (S4), Respiratory Control Ratio (RCR) and Swelling (S). Liver enzymes (AST, ALT, LDH) were determined in solution. At tissue, Malondialdehyde (MDA) and Nitrate (NOx) were determined. All parameters were analyzed at 0.6 and 24 hours of hypothermic preservation. Statistics analysis were made by Mann-Whitney test (p<0.05). Results Regarding ALT, there was a difference between FBP-6h/HTK-6h, lower in HTK. Regarding AST, there was a significant difference between FBP-24h/HTK-24h, lower in FBP. Regarding NOx, there was a difference between 0h and 6h, as well as 0h and 24h for both solutions. Regarding S3, there was a significant difference in 24h compared to Control-0h for both solutions, and a significant difference between FBP-6h/FBP-24h. Regarding S4, there was a difference between Control-0h/HTK-24h and FBP-24h/HTK-24h, higher in HTK. There was a difference between Control-0h/FBP-24h for Swelling, higher in FBP. Conclusion Fructose-1,6-Bisphosphate showed better performance at nitrate and aspartate aminotransferase compared to histidine-tryptophan-ketoglutarate.

Asunto(s)

Isquemia Fría , Alopurinol , Animales , Fructosa , Glucosa , Glutatión , Histidina , Hígado , Masculino , Manitol , Preservación de Órganos , Soluciones Preservantes de Órganos , Ratas , Ratas Sprague-Dawley , Triptófano

RESUMEN

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.

Asunto(s)

Glutatión/química , Soluciones Preservantes de Órganos/química , Trasplante de Órganos/métodos , Calcio/análisis , Humanos , Soluciones Preservantes de Órganos/normas , Temperatura , Factores de Tiempo

RESUMEN

BACKGROUND/AIMS: Dental avulsion is defined as the complete displacement of a tooth from its socket owing to trauma. The treatment outcome depends on storage of the avulsed teeth in media capable of maintaining the viability of periodontal ligament cells, when immediate replantation is not possible. To maintain the viability of periodontal ligament cells, plants can be used as a storage medium because of their pharmacological and phytotherapic properties. The aim of this study was to evaluate the effect of plants on the tissue repair following tooth replantation. METHODS: This systematic review was conducted according to the PRISMA guidelines and included articles collected in the Cochrane, LILACS, PubMed, Science Direct, Scopus and Web of Science databases, plus articles found in the grey literature. The articles were screened for partial reading using the Endnote and Rayyan platform. The methodology of studies was evaluated by using the OHAT and GRADE. RESULTS: In the initial search, 2361 articles were obtained, only 51 articles were submitted to complete reading, and 35 articles were selected for the qualitative analysis. The evaluated plants had a potential effect on cell viability and proliferation. The articles evaluated mainly the action of plants on cells of the periodontal ligament. Propolis, coconut water and Aloe vera were the most common storage medium. CONCLUSION: The methodological limitations persist, and the evaluation of the pharmacological potential of plants on dental tissues still requires more research.

Asunto(s)

Aloe , Cocos , Soluciones Preservantes de Órganos , Própolis , Avulsión de Diente , Humanos , Ligamento Periodontal , Reimplante Dental

RESUMEN

AIM: The purpose of this study was to investigate the effect of different storage media on viability and proliferation capacity of periodontal ligament cells. METHODS: Plates with periodontal ligament fibroblasts (PDLF) cells were incubated in skimmed and whole milk, recently prepared Hank's balanced salt solution (HBSS), Save-A-Tooth system's, coconut water, propylene glycol with 20% propolis, egg white, tap water (negative control) at 5 °C and 20 °C, for 24 h. In one of the plates of each temperature, cell viability was determined by MTT assay. In the remaining plates, the wells were filled and incubated with Minimum Essential Medium (MEM) at 37 °C for 24, 48, 72, 96, and 120 h. The proliferation capacity of PDFL cells was also evaluated by MTT assay. Data were statistically analyzed by Kruskal-Wallis, Scheffé and Mann-Whitney tests (α = 5%). RESULTS: At 5 °C, milk maintained more viable cells than other storage media immediately after exposure (0 h) and allowed greater proliferation capacity. At 20 °C, milk and HBSS had similar and allowed similar proliferation ability at 24 and 48 h. From 72 h onwards, capacity to maintain cell viability the proliferation rate of cells incubated in HBSS was superior than milk. At both temperature and experiments, Save-A-Tooth system was similar to tap water. CONCLUSION: Milk and HBSS were more effective in maintaining cellular viability and proliferation capacity than any other storage media. At 5 °C, the most viable alternative was milk. At 20 °C, HBSS had better results.

Asunto(s)

Soluciones Preservantes de Órganos , Avulsión de Diente , Animales , Proliferación Celular , Supervivencia Celular , Células Cultivadas , Medios de Cultivo , Humanos , Soluciones Isotónicas , Leche , Ligamento Periodontal

RESUMEN

BACKGROUND: Compromised microvasculature resulting from disrupted bronchial arterial circulation appears to trigger chronic lung allograft dysfunction. Maintaining the microvasculature throughout the transplant process could improve the long-term health of transplanted lungs. We recently developed a bronchial-arterial-circulation-sparing (BACS) lung preservation approach and tested whether this approach would decrease microvascular damage and improve allograft function. METHODS: The lungs of Lewis rats were procured using either the BACS approach, where the bronchial and pulmonary arteries were synchronously perfused; a conventional approach, where only the pulmonary artery was perfused; or a conventional approach with a prostaglandin flush. After 4 hours of cold ischemia, physiologic function and vascular tone of the grafts were evaluated during ex vivo lung perfusion (EVLP), and microvasculature damage was assessed using 2-photon microscopy analysis. Lung function was compared after transplant among the groups. RESULTS: After 4 hours of cold ischemia, the BACS group exhibited significantly higher adenosine triphosphate levels and lower expression of phosphorylated myosin light chain, which is essential for vascular smooth muscle contraction. On EVLP, the BACS and prostaglandin groups showed lower pulmonary vascular resistance and less arterial stiffness. BACS attenuated microvasculature damage in the lung grafts when compared with conventional preservation. After transplantation, the lungs preserved with the BACS approach exhibited significantly better graft function and lower expression of phosphorylated myosin light chain. CONCLUSIONS: Our data suggest that BACS lung preservation protects the dual circulation inherent to the lungs, facilitating robust microvasculature in lung grafts after transplantation, leading to better posttransplant outcomes.

Asunto(s)

Rechazo de Injerto/prevención & control , Trasplante de Pulmón/efectos adversos , Perfusión/métodos , Aloinjertos/irrigación sanguínea , Aloinjertos/patología , Animales , Bronquios/irrigación sanguínea , Bronquios/patología , Arterias Bronquiales/patología , Arterias Bronquiales/trasplante , Modelos Animales de Enfermedad , Circulación Extracorporea/instrumentación , Circulación Extracorporea/métodos , Rechazo de Injerto/patología , Humanos , Trasplante de Pulmón/métodos , Masculino , Microvasos/patología , Preservación de Órganos , Soluciones Preservantes de Órganos , Perfusión/instrumentación , Neumonectomía/métodos , Arteria Pulmonar/patología , Arteria Pulmonar/trasplante , Ratas , Ratas Endogámicas Lew , Obtención de Tejidos y Órganos/métodos , Isquemia Tibia/efectos adversos

RESUMEN

Resumo Contexto A isquemia e reperfusão (I/R) renal está envolvida diretamente com insuficiência renal aguda, ocorrendo em casos como infarto por embolização ou trombose, quadros de septicemia e transplante renal. Esse processo é complexo, envolvendo respostas imunes inatas e adaptativas, presença de infiltrado celular, produção e liberação de citocinas e quimiocinas. Também desencadeia respostas celulares e liberação de espécies reativas de oxigênio, além de resultar em apoptose e, em alguns casos, necrose celular. Nesse contexto, é imprescindível a avaliação dos mecanismos de proteção ao tecido renal. Objetivos O objetivo foi testar a solução desenvolvida M&G, avaliando sua capacidade protetora no rim por meio de análise morfométrica e presença e expressão de citocinas inflamatórias (TNF-alfa, VEGF, HIF e IL-8). Métodos Foram selecionados 18 ratos Wistar, divididos em três grupos: Sham (S), Controle (C) e Estudo (E). O grupo S foi submetido ao processo cirúrgico sem o clampeamento arterial. No grupo C, foi clampeada a aorta acima e abaixo da artéria renal esquerda, sem a infusão de solução preservadora. No grupo E, além do clampeamento, realizou-se a punção da aorta e a infusão contínua da solução M&G por 20 minutos a 15 °C. Realizou-se a avaliação morfológica e imuno-histoquímica com os marcadores. Resultados Identificaram-se diferenças morfológicas entre o grupo S comparado aos grupos C e E. Na análise dos marcadores, houve redução na intensidade de expressão do TNF e na expressão do VEGF no grupo E. Não houve diferenças com HIF e IL-8 entre os grupos. Conclusões A solução M&G apresentou redução da presença e expressão de TNF-alfa e tendência de redução do VEGF.

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Abstract Background Renal ischemia-reperfusion (I/R) is directly associated with acute renal failure and can occur in conditions such as infarction caused by embolization or thrombosis, septicemia, and kidney transplantation. The process is complex, involving innate and adaptive immune responses, presence of cellular infiltrate, and production and release of cytokines and chemokines. It also triggers cell responses and release of reactive oxygen species, in addition to causing apoptosis and, in some cases, cell necrosis. Against this background, evaluation of renal tissue protection mechanisms is essential. Objectives The objective of this study was to test the M&G solution, developed in prior research, evaluating its capacity to protect the kidneys using morphometric analysis and by assaying the presence and expression of inflammatory cytokines (TNF-alpha, VEGF, HIF, and IL-8). Methods Eighteen Wistar rats were divided into three groups: Sham (S), Control (C), and Experimental (E). The S group underwent the surgical operation, but without arterial clamping. In group C, the aorta was clamped above and below the left renal artery, without infusion of the preservation solution. In group E, in addition to clamping, the aorta was punctured and M&G solution was infused continuously for 20 minutes at 15o C. Morphological analysis and immunohistochemical assessment of markers were then conducted. Results Morphological differences were identified in group S compared with groups C and E. Analysis of markers revealed reduced intensity of expression of TNF and of VEGF in group E. There were no differences in HIF or IL-8 between groups. Conclusions The M&G solution was associated with a reduction in presence and expression of TNF-alpha and a trend to reduced VEGF.

Asunto(s)

Animales , Masculino , Ratas , Reperfusión/métodos , Soluciones Preservantes de Órganos/uso terapéutico , Isquemia/complicaciones , Riñón , Fosfatos , Cloruro de Potasio , Cloruro de Sodio , Ratas Wistar , Bicarbonato de Sodio , Insuficiencia Renal/terapia

RESUMEN

The objective of this study was evaluate the maintenance of the corneal endothelium of horses in cold EUSOL-C® preservation medium over different periods (seven and 14 days) using scanning electron microscopy. A total of 20 pairs of eyes from horses were analysed. The corneas were divided into four groups of 10 corneas each (G1, G2, G3 and G4): G1 - the samples were kept in the preservation medium for seven days; G3 - the samples were kept in the preservation medium for for 14 days; G2 and G4 were formed by the control corneal buttons of G1 and G3, respectively. The average cell loss observed in G1 was 7.62%, in G2 it was 7.04%, in G3 9.12% and in G4 7.16%. No statistically significant differences were observed between the four groups. It was concluded that the Eusol-C® hypothermic preservation medium provided satisfactory preservation of the corneal endothelium in equine species for up to 14 days.

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Objetivou-se avaliar a manutenção do endotélio da córnea de equinos em meio de preservação a frio EUSOL-C® em diferentes períodos de acondicionamento (sete e 14 dias) utilizando microscopia eletrônica de varredura. Foram avaliados 40 bulbos oculares de 20 equinos. Os bulbos oculares foram divididos em quatro grupos (G1, G2, G3 e G4), nos quais: G1 foi composto por 10 botões corneais acondicionados em meio de preservação para córnea Eusol-C® durante sete dias; o G3 foi formado por 10 botões corneais acondicionados em meio de preservação para córnea Eusol-C® durante 14 dias. O G2 e G4 foram formados pelas córneas controle armazenadas imediatamente em glutaraldeído. A média da perda celular observada no G1 foi de 7,62%, no G2 foi de 7,04 %, no G3 foi de 9,12% e o no G4 foi de 7,16%. Não foram observadas diferenças estatisticamente significativas entre os quatro grupos. Foi possível concluir que o meio de preservação hipotérmico Eusol-C® proporcionou de forma satisfatória a preservação do endotélio da córnea na espécie equina durante o período de até 14 dias.

Asunto(s)

Animales , Caballos , Criopreservación/veterinaria , Soluciones Preservantes de Órganos , Trasplante de Córnea/veterinaria , Endotelio

RESUMEN

The objective of this study was evaluate the maintenance of the corneal endothelium of horses in cold EUSOL-C® preservation medium over different periods (seven and 14 days) using scanning electron microscopy. A total of 20 pairs of eyes from horses were analysed. The corneas were divided into four groups of 10 corneas each (G1, G2, G3 and G4): G1 - the samples were kept in the preservation medium for seven days; G3 - the samples were kept in the preservation medium for for 14 days; G2 and G4 were formed by the control corneal buttons of G1 and G3, respectively. The average cell loss observed in G1 was 7.62%, in G2 it was 7.04%, in G3 9.12% and in G4 7.16%. No statistically significant differences were observed between the four groups. It was concluded that the Eusol-C® hypothermic preservation medium provided satisfactory preservation of the corneal endothelium in equine species for up to 14 days.(AU)

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Objetivou-se avaliar a manutenção do endotélio da córnea de equinos em meio de preservação a frio EUSOL-C® em diferentes períodos de acondicionamento (sete e 14 dias) utilizando microscopia eletrônica de varredura. Foram avaliados 40 bulbos oculares de 20 equinos. Os bulbos oculares foram divididos em quatro grupos (G1, G2, G3 e G4), nos quais: G1 foi composto por 10 botões corneais acondicionados em meio de preservação para córnea Eusol-C® durante sete dias; o G3 foi formado por 10 botões corneais acondicionados em meio de preservação para córnea Eusol-C® durante 14 dias. O G2 e G4 foram formados pelas córneas controle armazenadas imediatamente em glutaraldeído. A média da perda celular observada no G1 foi de 7,62%, no G2 foi de 7,04 %, no G3 foi de 9,12% e o no G4 foi de 7,16%. Não foram observadas diferenças estatisticamente significativas entre os quatro grupos. Foi possível concluir que o meio de preservação hipotérmico Eusol-C® proporcionou de forma satisfatória a preservação do endotélio da córnea na espécie equina durante o período de até 14 dias.(AU)

Asunto(s)

Animales , Caballos , Soluciones Preservantes de Órganos , Criopreservación/veterinaria , Trasplante de Córnea/veterinaria , Endotelio

RESUMEN

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.

Asunto(s)

Humanos , Trasplante de Órganos/métodos , Soluciones Preservantes de Órganos/química , Glutatión/química , Temperatura , Factores de Tiempo , Calcio/análisis , Soluciones Preservantes de Órganos/normas

RESUMEN

Abstract Purpose To compare Fructose-1,6-Bisphosphate (FBP) to Histidine-Tryptophan-Ketoglutarate (HTK) in liver preservation at cold ischemia. Methods Male rats (Sprague-Dawley: 280-340g) divided into three groups (n=7): Control; Fructose-1,6-bisphosphate (FBP); Histidine-Tryptophan-Ketoglutarate (HTK). Animals underwent laparotomy-thoracotomy for perfusion of livers with saline. Livers were removed and deposited into solutions. Mitochondria were isolated to determine State 3 (S3), State 4 (S4), Respiratory Control Ratio (RCR) and Swelling (S). Liver enzymes (AST, ALT, LDH) were determined in solution. At tissue, Malondialdehyde (MDA) and Nitrate (NOx) were determined. All parameters were analyzed at 0.6 and 24 hours of hypothermic preservation. Statistics analysis were made by Mann-Whitney test (p<0.05). Results Regarding ALT, there was a difference between FBP-6h/HTK-6h, lower in HTK. Regarding AST, there was a significant difference between FBP-24h/HTK-24h, lower in FBP. Regarding NOx, there was a difference between 0h and 6h, as well as 0h and 24h for both solutions. Regarding S3, there was a significant difference in 24h compared to Control-0h for both solutions, and a significant difference between FBP-6h/FBP-24h. Regarding S4, there was a difference between Control-0h/HTK-24h and FBP-24h/HTK-24h, higher in HTK. There was a difference between Control-0h/FBP-24h for Swelling, higher in FBP. Conclusion Fructose-1,6-Bisphosphate showed better performance at nitrate and aspartate aminotransferase compared to histidine-tryptophan-ketoglutarate.

Asunto(s)

Animales , Ratas , Isquemia Fría , Preservación de Órganos , Triptófano , Alopurinol , Ratas Sprague-Dawley , Soluciones Preservantes de Órganos , Fructosa , Glucosa , Glutatión , Histidina , Hígado , Manitol

RESUMEN

This study analyzed the efficacy of autologous platelet-rich fibrin (PRF) in maintaining and recovering cell viability of the periodontal ligament (PDL). The PDL cells were isolated from 45 extracted teeth randomly distributed among 6 groups: 5 min, 1 h, 2 h, PRF 30 min, PRF 1 h and PRF 2 h. In the groups 5 min, 1 h and 2 h (n = 5), the teeth were kept dry in extra-alveolar times of 5 min, 1 h and 2 h respectively. The teeth of the groups PRF 30 min, PRF 1 h and PRF 2 h (n = 10) were kept dry at extra-alveolar times of 30 min, 1 and 2 h followed by immersion in PRF for 45 min. PDL cells were isolated by enzymatic digestion with type II collagenase and dispase, counted and analyzed for viability with Trypan blue vital dye in Neubauer chamber. The variables total number of cells and cell viability demonstrated that in the 5 min, 1 h and 2 h groups there was a decrease after the extra-alveolar dry times of 1 and 2 h. In comparison with the total number of cells, group 1 h, considered immediate reimplantation, did not present statistical difference when compared to the groups PRF 30 min, PRF 1 h and 2 h, a result that demonstrates that PRF assists in cell maintenance and recovery. PRF provided increased cell viability in relation to the different dry extra-alveolar times analyzed (p < 0.001). Autologous PRF presented effectiveness in maintaining and recovering PDL cells from extracted teeth and kept dry for up to 2 h.

Asunto(s)

Ligamento Periodontal/citología , Ligamento Periodontal/metabolismo , Fibrina Rica en Plaquetas/metabolismo , Supervivencia Celular/fisiología , Células Cultivadas , Colagenasas/metabolismo , Endopeptidasas/metabolismo , Humanos , Microscopía , Soluciones Preservantes de Órganos

RESUMEN

Although reports of infections caused by anaerobes after tissue transplantation are uncommon, contamination of allografts may result in substantial complications. Anaerobic incubation and testing of organ transport solution (TS) are not routine. The aim of this study was to determine the bioburden of strict anaerobic bacteria and oxygen tension of heart-TS. Forty TS from different donors were evaluated cultured using membrane filtration (MF), direct inoculation on broth and automated blood culture bottle (ABCB). Bacterial identification was performed by MALDI-TOF. The transport conditions were simulated to verify the bacterial recovery. A sterile bag fulfilled with 250 ml-1 of sterile saline was spiked with 100 CFU ml-1 of Clostridium perfringens and the fluid recovered 0 h, 1 h, 2 h, 6 h, 12 h, 24 h and 48 h for culture and oxygen measurement. Strict anaerobic bacteria were not isolated in heart-TS. The recovery of C.perfringens spiked in heart-TS was 100% using automated blood culture bottles. MF method detected >100 CFU only after 6 h of spiking. The manual culture was not able to recover C.perfringens after the process. The percentage of O2 measures varied from 77.6 to 87.9%. MF or ABCB are better than direct inoculation for recovery of anaerobes from heart-TS. Although all samples from heart donors were negative for anaerobes (probably due to low incidence of contamination), C.perfringens were all recovered in the simulated transport condition.

Asunto(s)

Aloinjertos/microbiología , Bacterias Anaerobias/aislamiento & purificación , Clostridium perfringens/aislamiento & purificación , Válvulas Cardíacas/microbiología , Válvulas Cardíacas/trasplante , Soluciones Preservantes de Órganos , Humanos