RESUMO
Score sheets were first introduced 30 years ago to assess pain, distress and suffering in animals. To date, however, there is still no general agreement on their use in research practice, and only a few publications can be found on this topic. In the present work, we demonstrate the use of a special score sheet for severity assessment in the first three postoperative days in two showcased studies performed on Wistar and Lewis rats undergoing liver resection or orthotopic liver transplantation, respectively. Scoring of different criteria and the total score were evaluated within each intervention. Additionally, both procedures were compared regarding their degree of severity. Suitability of these score sheets was evaluated for assessing severity of the procedures and these showed a minor severity within each investigated study. A comparison of both studies showed slightly higher scores involving liver transplantation. In contradiction to the common classification of these procedures as a moderate severity grade the score sheets applied here indicates a minor severity grade within each investigated study. Also, limitations and possible improvements in the design of our score sheets for defined interventions are reconsidered.
Assuntos
Bem-Estar do Animal , Hepatectomia/efeitos adversos , Medição da Dor/métodos , Projetos de Pesquisa , Animais , Transplante de Fígado/efeitos adversos , Masculino , Ratos , Ratos Endogâmicos Lew , Ratos WistarRESUMO
Although the recognition of pain, distress and discomfort has already been described in 1985 by Morton and Griffiths there is still very little known about the establishment of score sheets especially, regarding post-surgical pain and severity assessment for laboratory animals such as rabbits. In this paper we describe the estimation of severity and recovery status of 36 female New Zealand White rabbits (NZW) in a standardized liver resection model using two different adhesive treatments and one control group. Welfare was assessed at 3-4 consecutive days after surgery using a scoring system which included the following criteria: body weight, general state, clinical results, spontaneous behavior and clinical examination. Values could range from 0 to 20 where increasing values indicated increasing severity with a predefined humane endpoint for a score ≥20 points. Documented score points were almost exclusively a result of body weight loss, whereas clinical signs and general health status had no influence on the overall sum of points scored. Behavioral variation was solely observed postoperatively, within the first 24 h, with an average score ≤1. In contrast to the classification of a laparotomy as a moderate procedure in the EU Directive 2010/63 (annex VIII) the assessment herein presented showed a mild burden in all groups according to the scoring system used. The partial hepatectomy itself, as well as the adhesive treatment using either synthetic glue VIVO-107 or fibrin glue, were well tolerated.
Assuntos
Bem-Estar do Animal , Hepatectomia/efeitos adversos , Medição da Dor/métodos , Projetos de Pesquisa , Animais , Feminino , CoelhosRESUMO
The aging society has a deep impact on patient care in urology. The number of patients in need of partial or whole bladder wall replacement is increasing simultaneously with the number of cancer incidents. Therefore, urological research requires a model of bladder wall replacement in adult and elderly people. Two types of porcine collagen I/III scaffolds were used in vitro for comparison of cell growth of two different pig breeds at different growth stages. Scaffolds were characterised with scanning electron and laser scanning microscopy. Urothelial and detrusor smooth muscle cells were isolated from 15 adult Göttingen minipigs and 15 juvenile German Landrace pigs. Growth behaviour was examined in cell culture and seeded onto the collagen scaffolds via immunohistochemistry, two-photon laser scanning microscopy and a viability assay. The collagen scaffolds showed different structured surfaces which are appropriate for seeding of the two different cell types. Moisturisation of the scaffolds resulted in a change of the structure. Cell growth of German Landrace urothelial cells and smooth muscle cells was significantly higher than cell growth of the Göttingen minipig cells. Seeding of scaffolds with both cell types from both pig races was possible which could be shown by immunohistochemistry and two-photon laser scanning microscopy. Growth behaviour on the scaffolds was significantly increased for the German Landrace compared to Göttingen minipig. Nevertheless, seeding with the adult Göttingen minipig cells resulted in a closed layer on the surface and urothelial cells and smooth muscle cells showed increasing growth until day 14. The results show that these collagen scaffolds are adequate for the seeding with vesical cells. Moreover, they seem appropriate for the use as an in vitro model for the adult or elderly as the cells of the adult Göttingen minipig too, show good growth behaviour.
Assuntos
Colágeno/química , Músculo Liso/citologia , Engenharia Tecidual/instrumentação , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Animais , Proliferação de Células , Sobrevivência Celular , Feminino , Imuno-Histoquímica , Masculino , Microscopia Eletrônica de Varredura , Microscopia de Fluorescência , Miócitos de Músculo Liso/citologia , Sefarose/química , Especificidade da Espécie , Suínos , Porco Miniatura , Bexiga Urinária/patologia , Urotélio/metabolismoAssuntos
Modelos Animais de Doenças , Ciência dos Animais de Laboratório , Hepatopatias , Animais , União Europeia , Guias como Assunto , Humanos , Ciência dos Animais de Laboratório/normas , Ciência dos Animais de Laboratório/estatística & dados numéricos , Hepatopatias/etiologia , Hepatopatias/patologia , Hepatopatias/fisiopatologia , Camundongos , RatosRESUMO
The induction of experimental obstructive cholestasis is a reliable model for cholestatic liver diseases in rodents. Bile duct ligation (BDL) in mice provokes typical time-dependent morphological and structural changes in the liver, ranging from liver cell injury and elevated serum enzyme levels after several days, to a severe inflammatory response in the liver after 5-7 days, up to an advanced hepatic fibrosis as soon as three to four weeks after surgical ligation of the common biliary duct. Upon BDL induction, hepatic stellate cells become activated and transdifferentiate into myofibroblasts that produce extracellular matrix proteins such as collagen. In principle, the periportal fibrosis induced by BDL in rat livers is reversible. After the relief of a biliary obstruction, the liver has the capacity to revert to a nearly normal histological architecture and a fully normal biochemical function. When BDL surgery is performed by an experienced scientist, this model has very high reproducibility among all fibrotic models. All these factors corroborate the outstanding value of this model for basic and translational research in biomedicine and hepatology. Nevertheless, this model can result in significant variations when surgery is carried out by untrained personnel or when unconscious modifications are implemented that affect the quality of the intervention. A detailed protocol is provided here for the provision of reliable and reproducible BDL in mice.
Assuntos
Ductos Biliares/cirurgia , Colestase/etiologia , Modelos Animais de Doenças , Ciência dos Animais de Laboratório , Animais , Colestase/patologia , Colestase/fisiopatologia , Colestase/cirurgia , Guias como Assunto , Humanos , Ciência dos Animais de Laboratório/normas , Ligadura , CamundongosRESUMO
One possible symptom of overactive bladder (OAB) is urinary incontinence, which is generally considered to be an age-associated disease and which is rapidly increasing with demographic changes. Rodent models are commonly used for the investigation of lower urinary tract functions, although the use of these species has limitations in several translational aspects. In biomedical research and preclinical toxicology, Göttingen minipigs are used increasingly. But in urological research, only few data are available for Göttingen minipigs. To the best of our knowledge, this study is one of the first to provide reference data of micturition in female Göttingen minipigs. Micturition frequency and volumes were monitored and analyzed in five female Göttingen minipigs. Voided volume was 520 ± 383 mL (mean ± standard deviation of mean [SD]) and micturition frequency 6.17 ± 3.68 (mean ± SD). We also performed a review of the literature to compare our data with data from different species (humans, pigs, rats and mice). Our findings revealed that micturition volume and frequency of Göttingen minipigs are more comparable with that of humans, leading to the conclusion that Göttingen minipigs may be the better choice for translational research in different research fields, such as urology, neurology and nephrology, etc. The provision of in vivo reference values meets with the 3R concept of 'reduction, refinement and replacement' of laboratory animals, because they allow comprehensive statistical power calculations (reduction), planning of telemetric approaches (refinement), and generation of computer-based modulation for the development of intravesical drug delivery systems (replacement).
Assuntos
Porco Miniatura/fisiologia , Micção , Animais , Feminino , Humanos , Camundongos/fisiologia , Ratos/fisiologia , Valores de Referência , Suínos , Urina/fisiologiaRESUMO
The rat is the most widely used animal species in surgical research and offers distinct advantages over the mouse in transplantation models due to its size and close genetic similarity to humans. Sequencing of the rat genome and successful application of transgenic technologies which had only been available for mice have since led to a resurgence of the use of rat models. Transplantation provides the possibility to deliver transgenes through a variety of routes which can potentially offer treatment modalities for post-transplant dysfunction and rejection. Moreover, the use of genetically encoded fluorescent light probes has enabled in vivo visualization of organs and tissue in living animals. In recent years, generation of gene knockout rats via the zinc-finger nuclease (ZFN) and transcription activator-like effector nuclease (TALEN) technologies has offered alternatives to the sophisticated embryonic stem cell based gene-targeting. In this review, we aim to provide an overview of transplantation studies involving transgenic techniques using rat models and recent advances in methods to modify the rat genome. Through novel gene modification techniques, precise, complete and conditional knockout and knockin rat models have become available which can provide promising new treatment options and opportunities for studying human transplant-related pathophysiology.
Assuntos
Marcação de Genes , Transplante de Órgãos , Ratos Transgênicos , Animais , Técnicas de Inativação de Genes , Marcação de Genes/métodos , Técnicas de Transferência de Genes , Transplante de Rim , Ratos , Traumatismo por Reperfusão/genética , Pesquisa , Ativação TranscricionalRESUMO
INTRODUCTION: Hernia repair with prosthetic meshes represents one of the most common surgical procedures in the field of surgery. This intervention is always associated with an ensuing inflammatory response, angiogenesis and fibrotic encapsulation forming a foreign body granuloma (FBG) around the mesh fibres. Several studies have described this inflammatory reaction by characterising inflammatory cell infiltrate around the FBG after mesh explantation. However, very little is known about the real-time progression of such an inflammatory response. The aim of this study was to investigate the feasibility of monitoring the ongoing inflammatory response to mesh implantation using bioluminescence in vivo. MATERIALS AND METHODS: Three luciferase transgenic mice strains (FVB/N-Tg(Vegfr2-luc)-Xen, BALB/C-Tg(NFκB-RE-luc)-Xen and Tg(INS/EpRE-Luc)T20Rbl) were used. Mice were anaesthetized with 2 % isoflurane, and two incisions were made on the left and right sides of the abdomen of the mice. A 1-cm(2) propylene mesh was implanted subcutaneously in the right incision wound of each mouse, and the left wound served as control. Two hundred microliters of D-luciferin was injected into the mice, and bioluminescence measurements were done prior to the surgical intervention and subsequently every 3 days. After mesh explantation, histological analysis was done. Statistical analysis was done using prism GraphPad software. RESULTS: Bioluminescence results revealed different time points of maximum signal for the different mice strains. VEGFR2 gene expression peaked on day 6, NFkB on day 12 and ARE on day 3 post mesh implantation. We also observed much higher bioluminescent signal around the FBG surrounding the mesh as compared to the control wound, with p < 0.05 for all the different mice strains. CONCLUSION: Our results prove the possibility of monitoring the inflammatory reaction after mesh implantation in vivo using bioluminescence signal release. This provides a novel method of accessing and accurately describing the ongoing inflammatory response over a given period of time.
Assuntos
Benzotiazóis , Reação a Corpo Estranho/patologia , Implantação de Prótese/efeitos adversos , Telas Cirúrgicas/efeitos adversos , Animais , Biópsia por Agulha , Modelos Animais de Doenças , Imuno-Histoquímica , Inflamação/patologia , Mediadores da Inflamação/sangue , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Transgênicos , Implantação de Prótese/métodos , Distribuição Aleatória , Medição de Risco , Estatísticas não Paramétricas , Cicatrização/fisiologiaRESUMO
Others and we have shown in several studies that the natural tetrahydropyrimidine ectoine protects mammalian cells and tissues against various stress factors including ischemia/reperfusion injury, UV-irradiation, and inflammation. Since little is known about the molecular mechanism of this protective effect, which was ascribed exclusively to an extracellular action of this small water-soluble molecule, we asked whether and how a hydrophobic anchor modulates the inflammation protective properties of ectoine. We therefore investigated the influence of ectoine and of its semi-synthetic derivative lauryl-ectoine on inflammation in RAW 264.7 macrophages and primary cultured rat intestinal smooth muscle (RISM) cells. Both, ectoine and lauryl-ectoine considerably decreased lipopolysaccharide (LPS)-induced interleukin (IL)- 1, IL-6, tumor necrosis factor (TNF)- α, and cyclooxygenase (COX)-2 gene expression in macrophages as well as TNF-α- induced IL-1, IL-6 and COX-2 expression in RISM cells. This reduction of inflammatory agents was accompanied on the one hand by a significant decrease of nuclear translocation of nuclear factor (NF)-κB and on the other hand by a reduction of cellular ceramide content. Interestingly, lauryl- ectoine was much more active exerting its effect at about 10-fold lower concentrations than its natural counterpart. Note that ectoine was almost completely recovered in the medium whereas lauryl-ectoine was found to be cell-associated. Together our data indicate that a lipid anchor considerably improves a possible preventive and/or therapeutic implementation of ectoine in inflammatory processes.
Assuntos
Diamino Aminoácidos/farmacologia , Diamino Aminoácidos/química , Animais , Linhagem Celular , Ciclo-Oxigenase 2/imunologia , Expressão Gênica , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Interleucina-1/imunologia , Interleucina-6/imunologia , Metabolismo dos Lipídeos/efeitos dos fármacos , Macrófagos/imunologia , Camundongos , NF-kappa B/imunologia , Ratos , Fator de Necrose Tumoral alfa/imunologiaRESUMO
The prognosis for recipients of small liver grafts is poor. The aim of this study was to determine the impact of venous systemic oxygen persufflation (VSOP) with nitric oxide (NO) gas for 30% partial liver preservation and transplantation in rats. After we determined optimal NO concentration as 40 ppm in vitro with the isolated perfused rat liver model, we assessed liver injury and regeneration in vivo at 1, 3, 24 and 168 h after transplantation in the following three groups after 3 h-cold storage (n = 20 per group): control group = static storage; VSOP group = oxygen persufflation and VSOP+NO group = oxygen with NO persufflation. The liver graft persufflation was achieved with medical gas via the suprahepatic vena cava; In comparison with control group after transplantation, VSOP+NO preservation (1) increased portal circulation, (2) reduced AST and ALT release, (3) upregulated hepatic endothelial NO synthase, (4) reduced hepatocyte and bileductule damage and (5) improved liver regeneration. These results suggest that gaseous oxygen with NO persufflation is a novel and safe preservation method for small partial liver grafts, not only alleviating graft injury but also improve liver regeneration after transplantation.
Assuntos
Transplante de Fígado , Óxido Nítrico/administração & dosagem , Preservação de Órgãos , Oxigênio/administração & dosagem , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , L-Lactato Desidrogenase/sangue , Regeneração Hepática , Microcirculação , Microscopia Eletrônica , Óxido Nítrico/análise , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo III/genética , RNA Mensageiro/genética , Ratos , Ratos Endogâmicos Lew , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Necrose Tumoral alfa/metabolismoRESUMO
The impact of surgical staplers on tissues has been studied mostly in an empirical manner. In this paper, finite element method was used to clarify the mechanics of tissue stapling and associated phenomena. Various stapling modalities and several designs of circular staplers were investigated to evaluate the impact of the device on tissues and mechanical performance of the end-to-end colorectal anastomosis. Numerical simulations demonstrated that a single row of staples is not adequate to resist leakage due to non-linear buckling and opening of the tissue layers between two adjacent staples. Compared to the single staple row configuration, significant increase in stress experienced by the tissue at the inner staple rows was observed in two and three rows designs. On the other hand, adding second and/or third staple row had no effect on strain in the tissue inside the staples. Variable height design with higher staples in outer rows significantly reduced the stresses and strains in outer rows when compared to the same configuration with flat cartridge.
Assuntos
Intestino Grosso/cirurgia , Modelos Biológicos , Grampeadores Cirúrgicos , Anastomose Cirúrgica/métodos , Desenho de Equipamento , Análise de Elementos Finitos , Estresse Mecânico , Grampeamento Cirúrgico/métodosRESUMO
The creation of musculoskeletal tissue represents an alternative for the replacement of soft tissue in reconstructive surgery. However, most of the approaches of creating artificial tissue have their limitations in the size as the maximally obtainable dimension of bioartificial tissue (BAT) is limited due to the lack of supporting vessels within the 3-dimensional construct. The seeded myoblasts require high amounts of perfusion, oxygen, and nutrients to survive. To achieve this, we developed a 3-dimensional scaffold which features the epigastric artery as macroscopic core vessel inside the BAT in a rat model (perfused group, n = 4) and a control group (n = 3) without the epigastric vessels and, therefore, without perfusion. The in vivo monitoring of the transplanted myoblasts was assessed by bioluminescence imaging and showed both the viability of the epigastric artery within the 3-dimensional construct and again that cell survival in vivo is highly depending on the blood supply with the beginning of capillarization within the BAT seven days after transplantation in the perfused group. However, further studies focussing on the matrix improvement will be necessary to create a transplantable BAT with the epigastric artery as anastomosable vessel.
RESUMO
BACKGROUND/AIMS: Chronic organ donor shortage has led to the consideration to expand the donor pool with livers from non-heart-beating donors (NHBD), although a higher risk of graft dys- or nonfunction is associated with these livers. We examined the effects of selective cyclooxygenase-2 (COX-2) inhibition on hepatic warm ischemia (WI) reperfusion (I/R) injury of NHBD. METHODS: Male Wistar rats were used as donors and meloxicam (5 mg/kg body weight) was administered into the preservation solution. Livers were excised after 60 min of WI in situ, flushed and preserved for 24 h at 4°C. Reperfusion was carried out in vitro at a constant flow for 45 min. During reperfusion (5, 15, 30 and 45 min), enzyme release of alanine aminotransferase and glutamate lactate dehydrogenase were measured as well as portal venous pressure, bile production and oxygen consumption. The production of malondialdehyde was quantified and TUNEL staining was performed. Quantitative PCR analyzed COX-2 mRNA. COX-2 immunohistochemistry and TxB(2) detection completed the measurements. RESULTS: Meloxicam treatment led to better functional recovery concerning liver enzyme release, vascular resistance and metabolic activity over time in all animals. Oxidative stress and apoptosis were considerably reduced. CONCLUSION: Cold storage using meloxicam resulted in significantly better integrity and function of livers retrieved from NHBD. Selective COX-2 inhibition is a new therapeutic approach achieving improved preservation of grafts from NHBD.
Assuntos
Inibidores de Ciclo-Oxigenase 2/farmacologia , Fígado/efeitos dos fármacos , Fígado/lesões , Traumatismo por Reperfusão/prevenção & controle , Tiazinas/farmacologia , Tiazóis/farmacologia , Animais , Apoptose/efeitos dos fármacos , Ciclo-Oxigenase 2/genética , Humanos , Fígado/fisiopatologia , Transplante de Fígado/métodos , Transplante de Fígado/fisiologia , Masculino , Meloxicam , Preservação de Órgãos/métodos , Estresse Oxidativo/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Traumatismo por Reperfusão/patologia , Traumatismo por Reperfusão/fisiopatologia , Doadores de Tecidos , Resistência Vascular/efeitos dos fármacosRESUMO
In order to reduce the number of animal experiments, the use of non-heart-beating donors (NHBDs) from a commercial abattoir has been proposed. Since the use of slaughterhouse organs is legally not defined as animal experiment, this would fulfil international standards as an alternative to animal experiments. The development of intravascular thrombosis after cardiac arrest negatively impacts organ preservation and thus viability during ischaemic storage and reperfusion. A fibrinolytic preflush with streptokinase might overcome these limitations. Therefore, the functional and histomorphological integrity of kidneys preserved immediately with intact circulation (control group A) and kidneys preserved after cardiac arrest with a 30 min period of warm ischaemia (WI) (group B) was compared with kidneys preflushed with 12.5 kU/L (group C) or 50 kU/L streptokinase (group D) after 30 min WI prior to preservation. We could demonstrate that kidneys preflushed with 12.5 kU/L streptokinase (group C) performed better than those without streptokinase pretreatment after WI (group B). Parameters like oxygen consumption, perfusion pressure, laboratory values, lactate dehydrogenase level and lipidperoxidation were closer to that of the control (group A) than in groups B and D. The higher streptokinase concentration of 50 kU/L (group D) resulted histologically in a more pronounced tissue damage and an attenuated renal function, indicating toxic effects. On the basis of our results we believe streptokinase preflushed slaughterhouse kidneys to be an adequate alternative to organs from laboratory animals with the potential to further reduce the number of animal experiments.
Assuntos
Fibrinolíticos/farmacologia , Preservação de Órgãos/métodos , Traumatismo por Reperfusão/prevenção & controle , Estreptoquinase/farmacologia , Isquemia Quente/veterinária , Alternativas ao Uso de Animais , Animais , Feminino , Parada Cardíaca , Rim/irrigação sanguínea , Rim/patologia , Testes de Função Renal/métodos , Soluções para Preservação de Órgãos , Perfusão , Sus scrofaRESUMO
To assess the effect of the perfusion pressure (PP) during machine perfusion (MP) on the preservation quality of kidney grafts, we compared mean PPs of 25 and 30 mmHg using a porcine autotransplantation model. After assessment of the microcirculation, animals underwent left nephrectomy. Thereafter, kidneys were washed out followed by 20 h of MP at 25 mmHg (MP25, n = 7) or 30 mmHg (MP30, n = 7) using a novel MP system for hypothermic pulsatile perfusion. After MP preservation, the contralateral kidneys were removed and the preserved kidneys heterotopically autotransplanted. Ten minutes after reperfusion, the microcirculation was reassessed. Seven days posttransplant, animals were euthanized and the kidney grafts removed for histological analysis. MP using a mean PP of 25 mmHg resulted in higher capillary blood flow after reperfusion. In the MP30 group, 6 out of 7 animals survived, whereas in the MP25 group all animals survived. Overall, improvement in recovery of renal function and a better preservation of structural integrity were seen in the MP25 group compared to the MP30 group. Using a novel system for hypothermic MP, a mean PP of 25 mmHg is preferred over a mean PP of 30 mmHg.
Assuntos
Hipotermia Induzida/métodos , Transplante de Rim , Rim/fisiologia , Rim/cirurgia , Preservação de Órgãos/métodos , Perfusão/métodos , Animais , Pressão Sanguínea , Suínos , Resultado do TratamentoRESUMO
BACKGROUND: Cold storage using histidine-tryptophan-ketoglutarate (HTK) solution is used widely in clinical practice for the preservation of warm ischaemia-damaged kidney grafts. This study assessed the efficacy of pulsatile machine perfusion in combination with Polysol for the preservation of warm ischaemia-damaged kidney grafts. METHODS: After induction of warm ischaemia by clamping of the left renal pedicle for 30 min, pigs were subjected to left nephrectomy. Thereafter, grafts were preserved for 20 h by cold storage with HTK (CS-HTK) or Polysol (CS-PS), or machine preservation with Polysol (MP-PS). Subsequently, contralateral kidneys were removed and preserved kidneys were transplanted. Control pigs underwent unilateral nephrectomy. Renal function was assessed daily for 1 week. Kidney biopsies were analysed for morphology and proliferative response. RESULTS: Renal function of warm ischaemia-damaged grafts preserved using MP-PS was comparable to that of non-ischaemic controls. MP-PS and CS-PS groups showed improved renal function compared with the CS-HTK group, with more favourable results for MP-PS than for CS-PS. The proliferative response of tubular cells in the CS-HTK group was higher than in all other groups. CONCLUSION: This study demonstrated that the function of warm ischaemia-damaged kidney grafts after pulsatile perfusion preservation was comparable to that of non-ischaemic controls.
Assuntos
Transplante de Rim/métodos , Rim/fisiologia , Soluções para Preservação de Órgãos/farmacologia , Isquemia Quente/métodos , Animais , Isquemia Fria/métodos , Constrição , Criopreservação/métodos , Glucose/administração & dosagem , Glucose/farmacologia , Imuno-Histoquímica , Rim/anatomia & histologia , Manitol/administração & dosagem , Manitol/farmacologia , Preservação de Órgãos/métodos , Soluções para Preservação de Órgãos/administração & dosagem , Tamanho do Órgão , Cloreto de Potássio/administração & dosagem , Cloreto de Potássio/farmacologia , Procaína/administração & dosagem , Procaína/farmacologia , Fluxo Pulsátil , Distribuição Aleatória , Ratos , Transplante AutólogoRESUMO
OBJECTIVE: The impact of different preservation solutions for washout of kidney grafts was evaluated regarding temperature, kidney weight, remaining red blood cells (RBCs) and histological evaluation after ex vivo washout using 500 mL cold preservation solution at 4 degrees C followed by 24 hours cold storage (CS). METHODS: Kidneys retrieved from Landrace pigs (20-30 kg) were immediately washed (warm ischemic time 0 min [WIT 0]), using 500 mL cold University of Wisconsin solution (UW), histidine-tryptophan-ketoglutarate (HTK), or Polysol (PS) followed by 24 hours, CS. Also, kidneys were retrieved after a WIT of 30 minutes followed by washout using HTK or PS. RESULTS: After washout, the weight of kidneys washed out with HTK had increased, whereas that of organs in the UW or PS group had decreased. After washout with UW, the core temperature of WIT 0 kidneys was lower than that with HTK. The time needed for washout using 500 mL solution was shorter using PS compared with HTK for both WIT 0 and WIT 30 groups. The amount of remaining RBCs was similar between all WIT 0 groups; whereas in the WIT 30 groups the amount was higher in kidneys washed out using HTK compared with PS. Histological evaluation showed less tissue injury among PS-washed kidneys compared with UW or HTK. CONCLUSION: Overall, kidneys washed-out with PS showed better preservation of structural integrity after 24 hours, CS compared with either UW or HTK. Washout of warm ischemically damaged kidneys was more effective using PS compared with HTK.
Assuntos
Transplante de Rim/fisiologia , Soluções para Preservação de Órgãos , Adenosina/química , Adenosina/farmacologia , Alopurinol/química , Alopurinol/farmacologia , Animais , Edema/patologia , Contagem de Eritrócitos , Glucose/química , Glucose/farmacologia , Glutationa/química , Glutationa/farmacologia , Insulina/química , Insulina/farmacologia , Rim/efeitos dos fármacos , Rim/patologia , Glomérulos Renais/patologia , Túbulos Renais/patologia , Manitol/química , Manitol/farmacologia , Preservação de Órgãos/métodos , Soluções para Preservação de Órgãos/química , Soluções para Preservação de Órgãos/farmacologia , Soluções para Preservação de Órgãos/uso terapêutico , Cloreto de Potássio/química , Cloreto de Potássio/farmacologia , Procaína/química , Procaína/farmacologia , Rafinose/química , Rafinose/farmacologia , Suínos , Doadores de TecidosRESUMO
BACKGROUND: Delayed graft function (DGF) is defined as posttransplantation dialysis within 1 week, which might be associated with impaired long-term graft survival. The aim of our pilot study was to establish the ability of intraoperative spectrometry of allograft microperfusion to predict DGF. METHODS: Twenty human kidney allografts transplanted from deceased donors were evaluated intraoperatively after reperfusion using modified organ spectrometry (O2C device). We examined hemoglobin oxygen saturation, intravascular amount of hemoglobin, and microperfusion flow/velocity. RESULTS: Retrospectively, 10/20 (50%) allografts with measurable impairment of cortical hemoglobin oxygen saturation and microperfusion flow/velocity developed DGF. Retrospectively, we found that if the intravascular amount of hemoglobin was increased upon intraoperative measurement, the kidney was prone to develop DGF. CONCLUSIONS: Spectrometry data predicted DGF. Our results supported the thesis that impaired microperfusion is the key to DGF and might be related to postcapillary endothelial damage or intravascular sludge.
Assuntos
Transplante de Rim/métodos , Transplante de Rim/fisiologia , Adulto , Pressão Sanguínea , Cadáver , Rejeição de Enxerto/epidemiologia , Hematoma/etiologia , Hematoma/terapia , Artéria Hepática/cirurgia , Veias Hepáticas/cirurgia , Humanos , Pessoa de Meia-Idade , Monitorização Intraoperatória/instrumentação , Monitorização Intraoperatória/métodos , Perfusão , Complicações Pós-Operatórias/epidemiologia , Valor Preditivo dos Testes , Estudos Retrospectivos , Stents , Doadores de Tecidos , Transplante Homólogo/métodos , Ureter/transplante , Bexiga Urinária/cirurgiaRESUMO
OBJECTIVE: We sought to assess the efficacy of POLYSOL, a low-viscosity, colloid-based organ preservation solution, for the preservation of warm ischemically damaged kidney grafts compared with histidine-tryptophane-ketoglutarate (HTK) solution. METHODS: Pigs (25-30 kg) underwent a left nephrectomy after clamping the renal vessels for 30 minutes. Kidney grafts washed out with Polysol (n = 6) or HTK (n = 6) were cold stored (CS) for 20 hours at 4 degrees C. After the preservation period, the contralateral kidney was removed and the preserved kidney implanted heterotopically. Renal function was assessed daily for 7 days. Thereafter, animals were killed and the kidney grafts removed for histologic analysis. RESULTS: All animals survived for 7 days. All Polysol CS-preserved grafts showed immediate function, as demonstrated by urine production within 24 hours after reperfusion as compared with 3/6 grafts in the HTK CS group. Overall, the Polysol CS group showed improved renal function compared with HTK CS. Also, peak serum creatinine and blood urea values were lower in the Polysol CS group compared with HTK-preserved grafts. Histologic evaluation of warm ischemically damaged grafts showed less glomerular shrinking, less tubular damage, less edema, less inflammatory infiltration, and less necrosis in Polysol compared with HTK-preserved grafts. CONCLUSION: Application of Polysol solution for washout and CS preservation of warm ischemically damaged kidney grafts resulted in improved renal function and structural integrity when compared with HTK.
Assuntos
Testes de Função Renal , Rim/patologia , Soluções para Preservação de Órgãos , Animais , Nitrogênio da Ureia Sanguínea , Creatinina/sangue , Diurese , Feminino , Glucose , Rim/efeitos dos fármacos , Rim/fisiologia , Manitol , Modelos Animais , Soluções para Preservação de Órgãos/farmacologia , Cloreto de Potássio , Procaína , Reperfusão , SuínosRESUMO
OBJECTIVE: To investigate the role of oxygen free radicals in the induction of apoptosis in non-heart-beating donor (NHBD) livers, and if superoxide dismutase (SOD) ameliorates these alterations. METHODS: Rat livers were perfused via the portal vein with histidine/tryptophan/alpha-ketoglutarate solution from heart-beating donors (HBD) or 60-min warm ischemia from NHBD, with or without the addition of SOD. After 24 h, cold storage livers were evaluated by isolated reperfusion. RESULTS: NHBD showed significantly higher enzyme leakage and elevated portal venous pressure (PVP) versus HBD. Bile and total adenine nucleotides (TAN) were significantly decreased. Apoptosis was prominent in sinusoidal lining cells, coupled with strong nitrotyrosine staining (NTR). The concentrations of nitric oxide and lipoperoxides were largely increased. SOD medication reduced hepatic enzyme release by 30% and lipoperoxides by nearly 50%. Apoptosis and NTR were significantly decreased, and PVP was strikingly reduced to normal values. A 3-fold enhancement in bile production and 1.5-fold increase in TAN of the liver tissue were also observed. CONCLUSION: NHBD livers are prone to severe reoxygenation injury promoted by oxygen free radicals, massive nitrite oxide production and peroxynitrite-induced apoptosis within the sinusoids. Antioxidant medication with SOD should be considered as a useful means of preserving NHBD livers.