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1.
Int J Mol Sci ; 25(17)2024 Aug 28.
Artículo en Inglés | MEDLINE | ID: mdl-39273266

RESUMEN

Liver transplantation remains the only definitive treatment for end-stage liver diseases. However, the increasing prevalence of fatty liver disease among potential donors exacerbates the shortage of suitable organs. This study evaluates the efficacy of the preservation solution Institut Georges Lopez-2 (IGL-2) compared to Histidine-Tryptophan-Ketoglutarate (HTK) and University of Wisconsin (UW) preservation solutions in mitigating ischemia-reperfusion injury (IRI) in steatotic livers. Using Zucker Obese rat livers, we assessed the impact of 24-h static cold storage (SCS) with each solution on transaminase release, glutathione redox balance, antioxidant enzyme activity, lipoperoxidation, and inflammation markers. IGL-2 and UW solutions demonstrated reduced transaminase and lactate levels compared to HTK, indicating better preservation of liver integrity. IGL-2 maintained a higher reduced glutathione/oxidized glutathione (GSH/GSSG) ratio, suggesting more effective management of oxidative stress. Antioxidant enzyme activities catalase, superoxide dismutase, and glutathione peroxidase (CAT, SOD, GPX) were higher in IGL-2 preserved livers, contributing to decreased oxidative damage. Lipid peroxidation markers and inflammatory markers were lower in IGL-2 than in HTK, indicating reduced oxidative stress and inflammation. Additionally, improved mitochondrial function was observed in the IGL-2 group, correlating with reduced reactive oxygen species (ROS) production and lipid peroxidation. These findings suggest that IGL-2 offers superior preservation of liver viability, reduces oxidative stress, and minimizes inflammation compared to HTK and UW solutions. By maintaining a higher ratio of reduced glutathione and antioxidant enzyme activity, IGL-2 effectively mitigates the harmful effects of ischemia-reperfusion injury. The reduced lipid peroxidation and inflammation in the IGL-2 group further underscore its potential in improving liver transplant outcomes. These results highlight the importance of optimizing preservation solutions to enhance the viability and functionality of donor organs, potentially expanding the donor pool and improving the success rates of liver transplantation. Future research should focus on refining preservation techniques and exploring additional protective agents to further improve organ preservation and transplant outcomes.


Asunto(s)
Adenosina , Alopurinol , Antioxidantes , Hígado Graso , Insulina , Hígado , Soluciones Preservantes de Órganos , Procaína , Rafinosa , Ratas Zucker , Daño por Reperfusión , Animales , Soluciones Preservantes de Órganos/farmacología , Ratas , Rafinosa/farmacología , Insulina/metabolismo , Adenosina/metabolismo , Adenosina/farmacología , Hígado Graso/metabolismo , Hígado Graso/tratamiento farmacológico , Hígado Graso/patología , Daño por Reperfusión/metabolismo , Daño por Reperfusión/tratamiento farmacológico , Daño por Reperfusión/prevención & control , Antioxidantes/farmacología , Antioxidantes/metabolismo , Hígado/metabolismo , Hígado/efectos de los fármacos , Hígado/patología , Alopurinol/farmacología , Masculino , Procaína/farmacología , Inflamación/metabolismo , Inflamación/patología , Inflamación/tratamiento farmacológico , Glucosa/metabolismo , Estrés Oxidativo/efectos de los fármacos , Glutatión/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Manitol/farmacología , Isquemia Fría/efectos adversos , Cloruro de Potasio/farmacología , Preservación de Órganos/métodos , Trasplante de Hígado/métodos
2.
Int J Mol Sci ; 25(17)2024 Sep 02.
Artículo en Inglés | MEDLINE | ID: mdl-39273476

RESUMEN

We recently reported in a rat model of kidney transplantation that the addition of sodium thiosulfate (STS) to organ preservation solution improved renal graft quality and prolonged recipient survival. The present study investigates whether STS pre-treatment would produce a similar effect. In vitro, rat kidney epithelial cells were treated with 150 µM STS before and/or during exposure to hypoxia followed by reoxygenation. In vivo, donor rats were treated with PBS or 2.4 mg/kg STS 30 min before donor kidneys were procured and stored in UW or UW+150 µM STS solution at 4 °C for 24 h. Renal grafts were then transplanted into bilaterally nephrectomised recipient rats which were then sacrificed on post-operative day 3. STS pre-treatment significantly reduced cell death compared to untreated and other treated cells in vitro (p < 0.05), which corresponded with our in vivo result (p < 0.05). However, no significant differences were observed in other parameters of tissue injury. Our results suggest that STS pre-treatment may improve renal graft function after transplantation.


Asunto(s)
Trasplante de Riñón , Riñón , Daño por Reperfusión , Tiosulfatos , Animales , Tiosulfatos/farmacología , Tiosulfatos/uso terapéutico , Daño por Reperfusión/tratamiento farmacológico , Trasplante de Riñón/efectos adversos , Trasplante de Riñón/métodos , Ratas , Masculino , Riñón/efectos de los fármacos , Soluciones Preservantes de Órganos/farmacología , Preservación de Órganos/métodos
3.
Transplant Proc ; 56(8): 1811-1819, 2024 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-39242310

RESUMEN

BACKGROUND: Reliable 24-hour preservation is required to optimize the rehabilitation potential of Ex Situ Lung Perfusion (ESLP). Other ESLP protocols include fresh perfusate replacement to counteract an accumulation of deleterious by-products. We describe the results of our reliable 24-hour negative pressure ventilation (NPV)-ESLP protocol with satisfactory acute post-transplant outcomes and investigate perfusate exchange (PE) as a modification to enhance prolonged ESLP. METHODS: Twelve pig lungs underwent 24 hours of NPV-ESLP using 1.5L of cellular perfusate (500 mL packed red blood cells and 1 L buffered perfusate). The Control (n = 6) had no PE; the PE (n = 6) had 500 mL replaced after 12 hours of NPV-ESLP with 1000 mL fresh perfusate. Three left lungs per group were transplanted. RESULTS: Results are reported as Control vs PE (mean ± SEM). Both groups demonstrated stable and acceptable oxygenation during 24 hours of ESLP with final PF ratios of 527.5 ± 42.19 and 488.4 ± 35.38 (P = .25). Final compliance measurements were 20.52 ± 3.59 and 18.55 ± 2.91 (P = .34). There were no significant differences in pulmonary artery pressure after 24 hours of ESLP (10.02 ± 2.69 vs 14.34 ± 1.64, P = .10), and pulmonary vascular resistance only differed significantly at T12 (417.6 ± 53.06 vs 685.4 ± 81.19, P = .02). Percentage weight gain between groups was similar (24.32 ± 8.4 and 45.33 ± 7.76, P = .07). Post-transplant left lung oxygenation was excellent (327.3 ± 14.62 and 313.3 ± 15.38, P = .28). There was no significant difference in % weight gain of lungs post-transplant (22.20 ± 7.22 vs 14.36 ± 9.96, P = .28). CONCLUSION: Acceptable lung function was maintained during 24-hour NPV-ESLP and post-transplant regardless of PE.


Asunto(s)
Trasplante de Pulmón , Pulmón , Perfusión , Animales , Perfusión/métodos , Porcinos , Preservación de Órganos/métodos , Factores de Tiempo , Soluciones Preservantes de Órganos
4.
Transplant Proc ; 56(8): 1732-1739, 2024 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-39242312

RESUMEN

OBJECTIVES: Ferroptosis plays a pivotal role in the pathogenesis of renal ischemia-reperfusion injury, where the processes are mediated by free ferrous ions and mitochondrial-released reactive oxygen species. However, the administration of high doses of cyclosporine A (CsA) or deferoxamine (DFO) poses a significant risk of renotoxicity. In contrast, low doses of DFO act as a ferrous iron chelator, and CsA functions as a mitochondrial reactive oxygen species blocker. This study aims to explore the potential protective effects of donor treatment with low-dose CsA, DFO, or their combination against ischemia-reperfusion injury during renal transplantation in a rat model. MATERIALS AND METHODS: In an ex vivo cold storage (CS) model utilizing renal slices, the impact of incorporating DFO, CsA, and a combination of both into the University of Wisconsin solution was assessed through the measurement of lactate dehydrogenase leakage. Additionally, their potential benefits were investigated in a rat donation after circulatory death (DCD) kidney transplant model, where the extent of damage was evaluated based on graft function, tubular necrosis, and inflammation. RESULTS: The co-administration of DFO and CsA effectively decreased the release of lactate dehydrogenase induced by CS ( P ≥ .05). In the in vivo model, this combined supplementation demonstrated a mitigating effect on reperfusion injury, evidenced by lower blood urea nitrogen levels and acute tubular necrosis scores compared to the control group (allP ≤ .05). Furthermore, the combined treatment significantly reduced apoptotic levels compared to the control group (P ≥ .05). CONCLUSIONS: The combined treatment with DFO and CsA mitigated the cold ischemia-reperfusion injury in the DCD kidney. Hence, this presents a new strategy for the CS of DCD kidney in clinical transplants.


Asunto(s)
Ciclosporina , Deferoxamina , Trasplante de Riñón , Daño por Reperfusión , Animales , Daño por Reperfusión/prevención & control , Daño por Reperfusión/patología , Daño por Reperfusión/etiología , Trasplante de Riñón/efectos adversos , Deferoxamina/farmacología , Ratas , Masculino , Riñón/patología , Riñón/efectos de los fármacos , Soluciones Preservantes de Órganos , Sinergismo Farmacológico , Isquemia Fría/efectos adversos , Preservación de Órganos/métodos , Modelos Animales de Enfermedad , Rafinosa/farmacología , Alopurinol
5.
Cryobiology ; 116: 104950, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-39134131

RESUMEN

Ischemia is a major limiting factor in Vascularized Composite Allotransplantation (VCA) as irreversible muscular injury can occur after as early as 4-6 h of static cold storage (SCS). Organ preservation technologies have led to the development of storage protocols extending rat liver ex vivo preservation up to 4 days. Development of such a protocol for VCAs has the added challenge of inherent ice nucleating factors of the graft, therefore, this study focused on developing a robust protocol for VCA supercooling. Rodent partial hindlimbs underwent subnormothermic machine perfusion (SNMP) with several loading solutions, followed by a storage solution with cryoprotective agents (CPA) developed for VCAs. Storage occurred in suspended animation for 24h and VCAs were recovered using SNMP with modified Steen. This study shows a robust VCA supercooling preservation protocol in a rodent model. Further optimization is expected to allow for its application in a transplantation model, which would be a breakthrough in the field of VCA preservation.


Asunto(s)
Criopreservación , Crioprotectores , Miembro Posterior , Preservación de Órganos , Alotrasplante Compuesto Vascularizado , Animales , Ratas , Miembro Posterior/irrigación sanguínea , Masculino , Alotrasplante Compuesto Vascularizado/métodos , Criopreservación/métodos , Preservación de Órganos/métodos , Crioprotectores/farmacología , Soluciones Preservantes de Órganos/farmacología , Perfusión/métodos , Aloinjertos Compuestos
6.
Transplant Proc ; 56(8): 1890-1895, 2024 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-39217028

RESUMEN

To resolve the critical donor shortage worldwide, enlarging the potential donor pool to include expanded criteria donors is necessary. Despite numerous attempts to establish new preservation solutions, no dramatic innovation has occurred since University of Wisconsin (UW) solution displaced Euro Collins' solution; UW solution remains the global gold standard. We previously developed a heavy water (D2O)-containing organ storage solution, Dsol, which is effective for livers subjected to extended cold storage (CS), and reported its effectiveness. Dsol is a modified UW solution; however, the substances or conditions that exhibit a synergistic or additive effect with D2O are unclear. Here we made UWD solution by removing hydroxyethyl starch (HES) from and adding 30%-D2O to UW solution, and compared the effects of these solutions. After 48 hours of CS, the livers were reperfused at 37 °C on an isolated perfused rat liver apparatus, and their perfusion kinetics, functions, and injuries were compared. In the UW group, portal vein resistance significantly increased and the oxygen consumption rate and bile production decreased; in contrast, these changes were suppressed in the UWD group. Organ expansion and liver damage progressed in both groups. These results confirmed that the removal of HES from and addition of D2O to the UW solution reduced CS-induced cellular function impairments and microcirculatory disorders. However, to reduce injury during reperfusion after CS, it is necessary to provide conditions that inhibit injury progression after reperfusion.


Asunto(s)
Adenosina , Alopurinol , Hígado , Soluciones Preservantes de Órganos , Preservación de Órganos , Rafinosa , Animales , Soluciones Preservantes de Órganos/farmacología , Hígado/efectos de los fármacos , Ratas , Preservación de Órganos/métodos , Rafinosa/farmacología , Alopurinol/farmacología , Adenosina/farmacología , Masculino , Insulina , Glutatión/farmacología , Ratas Sprague-Dawley , Óxido de Deuterio/farmacología , Trasplante de Hígado
7.
Medicina (Kaunas) ; 60(8)2024 Aug 09.
Artículo en Inglés | MEDLINE | ID: mdl-39202566

RESUMEN

Background and objectives: The main problem of vascular preservation is the maintenance of vessel graft quality and function following extended storage. Conventional preservation solutions such as histidine-tryptophan-ketoglutarate (HTK) solution, Phosphate-Buffer Solution (PBS), or sodium chloride 0.9% has been shown to be inadequate in preserving vascular physiological function after 3 days of cold storage. This study aimed to evaluate whether adenosine and lidocaine (AL) in a modified Krebs-Henseleit (KH) solution can preserve the function and histological structure of rat aortic rings after 6 days. Materials and Methods: Thirty-five aortic rings from male Wistar rats (200-300 g) were harvested and immediately immersed in one of the assigned cold preservation solutions: standard KH, modified KH (mod KH) with lower calcium (Ca2+) and higher magnesium content (Mg2+) with or without adenosine and lidocaine (mod KH-AL), and modified KH with AL, insulin, and melatonin (Mod KH-ALMI). The contraction and relaxation function of the aortic rings were examined using an isometric force transducer after 6 days of cold preservation. Hematoxylin and eosin staining were used to analyze the rings' histological structure. Results: Vascular contraction and relaxation functions were severely affected after a 6-day cold storage period in standard KH. Modifying the KH solution by reducing the Ca2+ and increasing the Mg2+ levels greatly recovered the vessel functions. The addition of AL or ALMI to the modified KH did not further recover vascular contractility. However, only the addition of AL to the modified KH increased the ACh-induced relaxation at 6 days when compared to the conventional KH, suggesting that endothelium preservation is improved. From histological analysis, it was found that the addition of AL but not ALMI further improved the endothelial lining and the structure of the elastic membrane layers of the preserved vessels after 6 days of cold preservation. Conclusions: The addition of AL to low calcium-high magnesium KH solution significantly enhanced endothelial preservation and improved endothelial-induced relaxation of preserved vessels after 6 days of cold storage.


Asunto(s)
Adenosina , Calcio , Lidocaína , Magnesio , Soluciones Preservantes de Órganos , Ratas Wistar , Animales , Lidocaína/farmacología , Adenosina/farmacología , Ratas , Masculino , Calcio/análisis , Magnesio/farmacología , Soluciones Preservantes de Órganos/farmacología , Aorta/efectos de los fármacos , Aorta/fisiología , Glucosa/farmacología , Cloruro de Potasio/farmacología , Trometamina
8.
Transpl Int ; 37: 11336, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38962471

RESUMEN

Segmental grafts from living donors have advantages over grafts from deceased donors when used for small intestine transplantation. However, storage time for small intestine grafts can be extremely short and optimal graft preservation conditions for short-term storage remain undetermined. Secreted factors from mesenchymal stem cells (MSCs) that allow direct activation of preserved small intestine grafts. Freshly excised Luc-Tg LEW rat tissues were incubated in preservation solutions containing MSC-conditioned medium (MSC-CM). Preserved Luc-Tg rat-derived grafts were then transplanted to wild-type recipients, after which survival, injury score, and tight junction protein expression were examined. Luminance for each graft was determined using in vivo imaging. The findings indicated that 30-100 and 3-10 kDa fractions of MSC-CM have superior activating effects for small intestine preservation. Expression of the tight-junction proteins claudin-3, and zonula occludens-1 preserved for 24 h in University of Wisconsin (UW) solution containing MSC-CM with 50-100 kDa, as shown by immunostaining, also indicated effectiveness. Reflecting the improved graft preservation, MSC-CM preloading of grafts increased survival rate from 0% to 87%. This is the first report of successful transplantation of small intestine grafts preserved for more than 24 h using a rodent model to evaluate graft preservation conditions that mimic clinical conditions.


Asunto(s)
Intestino Delgado , Células Madre Mesenquimatosas , Preservación de Órganos , Ratas Endogámicas Lew , Animales , Intestino Delgado/trasplante , Ratas , Preservación de Órganos/métodos , Masculino , Soluciones Preservantes de Órganos , Supervivencia de Injerto , Medios de Cultivo Condicionados , Proteína de la Zonula Occludens-1/metabolismo , Claudina-3/metabolismo , Ratas Transgénicas , Glutatión , Rafinosa , Alopurinol , Insulina , Adenosina
9.
J Vis Exp ; (208)2024 Jun 14.
Artículo en Inglés | MEDLINE | ID: mdl-38949382

RESUMEN

Lung transplantation is hampered by the lack of suitable donors. Previously, donors that were thought to be marginal or inadequate were discarded. However, new and exciting technology, such as ex vivo lung perfusion (EVLP), offers lung transplant providers extended assessment for marginal donor allografts. This dynamic assessment platform has led to an increase in lung transplantation and has allowed providers to use donors that were previously discarded, thus expanding the donor pool. Current perfusion techniques use cellular or acellular perfusates, and both have distinct advantages and disadvantages. Perfusion composition is critical to maintaining a homeostatic environment, providing adequate metabolic support, decreasing inflammation and cellular death, and ultimately improving organ function. Perfusion solutions must contain sufficient protein concentration to maintain appropriate oncotic pressure. However, current perfusion solutions often lead to fluid extravasation through the pulmonary endothelium, resulting in inadvertent pulmonary edema and damage. Thus, it is necessary to develop novel perfusion solutions that prevent excessive damage while maintaining proper cellular homeostasis. Here, we describe the application of a polymerized human hemoglobin (PolyhHb)-based oxygen carrier as a perfusate and the protocol in which this perfusion solution can be tested in a model of rat EVLP. The goal of this study is to provide the lung transplant community with key information in designing and developing novel perfusion solutions, as well as the proper protocols to test them in clinically relevant translational transplant models.


Asunto(s)
Hemoglobinas , Trasplante de Pulmón , Pulmón , Perfusión , Animales , Ratas , Trasplante de Pulmón/métodos , Hemoglobinas/química , Perfusión/métodos , Pulmón/metabolismo , Humanos , Oxígeno/metabolismo , Sustitutos Sanguíneos/farmacología , Sustitutos Sanguíneos/química , Masculino , Soluciones Preservantes de Órganos/química
10.
PLoS One ; 19(7): e0306056, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38968266

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
11.
Molecules ; 29(13)2024 Jul 03.
Artículo en Inglés | MEDLINE | ID: mdl-38999118

RESUMEN

In this study, a liquid chromatographic method was developed for the fast determination of lincomycin, polymyxin and vancomycin in a preservation solution for transplants. A Kinetex EVO C18 (150 × 4.6 mm, 2.6 µm) column was utilized at 45 °C. Gradient elution was applied using a mixture of mobile phases A and B, both including 30 mM phosphate buffer at pH 2.0 and acetonitrile, at a ratio of 95:5 (v/v) for A and 50:50 (v/v) for B. A flow rate of 1.0 mL/min, an injection volume of 20 µL and UV detection at 210 nm were used. A degradation study treating the three antibiotics with 0.5 M hydrochloric acid, 0.5 M sodium hydroxide and 3% H2O2 indicated that the developed method was selective toward lincomycin, polymyxin, vancomycin and their degradation products. Other ingredients of the preservation solution, like those from the cell culture medium, did not interfere. The method was validated with good sensitivity, linearity, precision and accuracy. Furthermore, lincomycin, polymyxin and vancomycin were found to be stable in this preservation solution for 4 weeks when stored at -20 °C.


Asunto(s)
Lincomicina , Polimixinas , Vancomicina , Lincomicina/análisis , Vancomicina/análisis , Polimixinas/análisis , Cromatografía Liquida/métodos , Soluciones Preservantes de Órganos , Antibacterianos/análisis , Reproducibilidad de los Resultados , Cromatografía Líquida de Alta Presión/métodos
12.
Transpl Int ; 37: 12994, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-39070247

RESUMEN

The aim of this study was to provide insight into high-energy phosphate compound concentration dynamics under realistic clinical cold-storage conditions using the Celsior solution in seven heart grafts discarded from transplantation. The hearts of seven local donors (three males, four females, age 37 ± 17 years, height 175 ± 5 cm, weight 75 ± 9 kg) initially considered for transplantation and eventually discarded were submitted to a Magnetic Resonance Spectroscopy observation in a clinical Magnetic Resonance Imaging scanner over at least 9 h. The grafts remained in their sterile container at 4°C during the entire examination. Hence, Phosphocreatine (PCr), adenosine triphosphate (ATP), inorganic phosphate (Pi) and intracellular pH were recorded non-destructively at a 30-minute interval. With the ischemic time Ti, the concentration ratios decreased at PCr/ATP = 1.68-0.0028·Tis, Pi/ATP = 1.38 + 0.0029·Tis, and intracellular pH at 7.43-0.0012·Tis. ATP concentration remained stable for at least 9 h and did not decrease as long as phosphocreatine was detectable. Acidosis remained moderate. In addition to the standard parameters assessed at the time of retrieval, Magnetic Resonance Spectroscopy can provide an assesment of the metabolic status of heart grafts before transplantation. These results show how HEPC metabolites deplete during cold storage. Although many parameters determine graft quality during cold storage, the dynamics of HEPC and intracellular pH may be helpful in the development of strategies aiming at extending the ischemic time.


Asunto(s)
Adenosina Trifosfato , Disacáridos , Electrólitos , Glutamatos , Glutatión , Trasplante de Corazón , Histidina , Manitol , Soluciones Preservantes de Órganos , Preservación de Órganos , Fosfatos , Humanos , Femenino , Masculino , Adenosina Trifosfato/metabolismo , Adulto , Persona de Mediana Edad , Preservación de Órganos/métodos , Espectroscopía de Resonancia Magnética , Concentración de Iones de Hidrógeno , Fosfocreatina/metabolismo , Adulto Joven , Criopreservación , Imagen por Resonancia Magnética
13.
Appl Opt ; 63(14): 3745-3752, 2024 May 10.
Artículo en Inglés | MEDLINE | ID: mdl-38856336

RESUMEN

The preservation method to store bone tissue for posterior analysis is a widespread practice. However, the method's potential influence on the material's mechanical properties is often overlooked during single-point experimentation. Saline and formaldehyde solutions are the most common among the employed preservation media. A full field analysis of the mice femoral bone deformation using non-destructive optical techniques is conducted to assess the influence of the storage media on the viscoelastic properties of the tissue. Three different groups are subjected to a standard three-point bending test. The first group is the control, with fresh post-mortem samples. The second and third groups used saline and formaldehyde solutions, respectively. During the mechanical test, the bone's surface and internal deformation are monitored simultaneously using digital holographic interferometry and Fourier-domain optical coherence tomography. A mechanical comparison among the three groups is presented. The results show that after 48 h of immersion in saline solution, the mice bones keep their viscoelastic behavior similar to fresh bones. Meanwhile, 48 h in formaldehyde modifies the response and affects the marrow structure. The high sensitivity of the optical phase also makes it possible to observe changes in the anisotropy of the samples. As a comparison, Raman spectroscopy analyzes the three bone groups to prove that the preservation media does not affect a single-point inspection.


Asunto(s)
Fémur , Formaldehído , Espectrometría Raman , Tomografía de Coherencia Óptica , Animales , Ratones , Formaldehído/farmacología , Tomografía de Coherencia Óptica/métodos , Espectrometría Raman/métodos , Fémur/diagnóstico por imagen , Fémur/fisiología , Pruebas Mecánicas , Elasticidad/efectos de los fármacos , Viscosidad , Soluciones Preservantes de Órganos/farmacología , Interferometría/métodos , Solución Salina
16.
Transpl Int ; 37: 12787, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38845758

RESUMEN

Organ quality can be assessed prior to transplantation, during normothermic machine perfusion (NMP) of the liver. Evaluation of mitochondrial function by high-resolution respirometry (HRR) may serve as a viability assessment concept in this setting. Freshly collected tissue is considered as optimal sample for HRR, but due to technical and personnel requirements, more flexible and schedulable measurements are needed. However, the impact of cold storage following NMP before processing biopsy samples for mitochondrial analysis remains unknown. We aimed at establishing an appropriate storage protocol of liver biopsies for HRR. Wedge biopsies of 5 human livers during NMP were obtained and assessed by HRR. Analysis was performed after 0, 4, 8, and 12 h of hypothermic storage (HTS) in HTK organ preservation solution at 4°C. With HTS up to 4 h, mitochondrial performance did not decrease in HTS samples compared with 0 h (OXPHOS, 44.62 [34.75-60.15] pmol·s-1·mg wet mass-1 vs. 43.73 [40.69-57.71], median [IQR], p > 0.999). However, at HTS beyond 4 h, mitochondrial respiration decreased. We conclude that HTS can be safely applied for extending the biopsy measurement window for up to 4 h to determine organ quality, but also that human liver respiration degrades beyond 4 h HTS following NMP.


Asunto(s)
Trasplante de Hígado , Hígado , Preservación de Órganos , Perfusión , Humanos , Preservación de Órganos/métodos , Hígado/patología , Biopsia , Masculino , Persona de Mediana Edad , Femenino , Mitocondrias Hepáticas/metabolismo , Soluciones Preservantes de Órganos , Anciano , Respiración de la Célula , Adulto
17.
Cryobiology ; 116: 104926, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-38880369

RESUMEN

Current methods of storing explanted donor livers at 4 °C in University of Wisconsin (UW) solution result in loss of graft function and ultimately lead to less-than-ideal outcomes post transplantation. Our lab has previously shown that supplementing UW solution with 35-kilodalton polyethylene glycol (PEG) has membrane stabilizing effects for cold stored primary rat hepatocytes in suspension. Expanding on past studies, we here investigate if PEG has the same beneficial effects in an adherent primary rat hepatocyte cold storage model. In addition, we investigated the extent of cold-induced apoptosis through treating cold-stored hepatocytes with pan caspase inhibitor emricasan. In parallel to storage at the current cold storage standard of 4 °C, we investigated the effects of lowering the storage temperature to -4 °C, at which the storage solution remains ice-free due to the supercooling phenomenon. We show the addition of 5 % PEG to the storage medium significantly reduced the release of lactate dehydrogenase (LDH) in plated rat hepatocytes and a combinatorial treatment with emricasan maintains hepatocyte viability and morphology following recovery from cold storage. These results show that cold-stored hepatocytes undergo multiple mechanisms of cold-induced injury and that PEG and emricasan treatment in combination with supercooling may improve cell and organ preservation.


Asunto(s)
Apoptosis , Inhibidores de Caspasas , Criopreservación , Hepatocitos , L-Lactato Deshidrogenasa , Soluciones Preservantes de Órganos , Polietilenglicoles , Animales , Hepatocitos/efectos de los fármacos , Polietilenglicoles/farmacología , Ratas , Soluciones Preservantes de Órganos/farmacología , Criopreservación/métodos , Masculino , L-Lactato Deshidrogenasa/metabolismo , Apoptosis/efectos de los fármacos , Inhibidores de Caspasas/farmacología , Rafinosa/farmacología , Células Cultivadas , Alopurinol/farmacología , Crioprotectores/farmacología , Frío , Glutatión/metabolismo , Glutatión/farmacología , Supervivencia Celular/efectos de los fármacos , Insulina/farmacología , Adenosina/farmacología , Preservación de Órganos/métodos , Ratas Sprague-Dawley , Ácidos Pentanoicos
18.
Cell Tissue Bank ; 25(3): 839-850, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-38822224

RESUMEN

In this experimental study, we compared the structural integrity and cell quality of corneal endothelium stored in organ culture medium (OCS) and Eusol-C. The experiment included rabbit and human cornea experiments in vitro. Thirty rabbit corneas and thirty-two human corneas were collected and divided into two groups. All right corneas were allocated in experiment group and left corneas were placed in control group. The corneas in experimental group were stored in OCS at 34 °C, and the corneas in control group were stored in Eusol-C at 4 °C for 7, 14, 21, 28, and 35 days, respectively. Endothelial cell morphology, cell count, and trypan blue staining for viability were assessed before storage (Day 0) and at days 7, 14, 21, 28 and 35. The structural integrity of human corneal endothelial cell was analyzed using immunohistochemistry. The samples of storage solution for microbial culture were collected on the third day and at the end of storage. The results show that no bacterial and fungal infections were found in both groups. After 14 days of storage, the morphology of endothelial cell was better in the experimental group than in the control group. The endothelial cell stored in OCS were better than those stored in Eusol-C at the end of storage times, except human cornea 14 days storage group. The ZO-1 protein staining showed the typical polygonal morphology of endothelial cell stored in the OCS. Corneal endothelial cells stored in the OCS had better quality up to 28 days. It can be applied to Chinese eye banks as a method of corneal preservation.


Asunto(s)
Endotelio Corneal , Técnicas de Cultivo de Órganos , Soluciones Preservantes de Órganos , Endotelio Corneal/citología , Animales , Conejos , Humanos , Soluciones Preservantes de Órganos/farmacología , Técnicas de Cultivo de Órganos/métodos , Preservación de Órganos/métodos , Recuento de Células , Masculino , Supervivencia Celular , Células Endoteliales/citología
19.
Cryobiology ; 115: 104904, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38734364

RESUMEN

Increasing shortage of donor organs leads to the acceptance of less than optimal grafts for transplantation, up to and including organs donated after circulatory standstill of the donor. Therefore, protective strategies and pharmacological interventions destined to reduce ischemia induced tissue injury are considered a worthwhile focus of research. The present study evaluates the potential of a multidrug pharmacological approach as single flush at the end of static preservation to protect the liver from reperfusion injury. Livers were retrieved from male Wistar rats 20 min after cardiac standstill. The organs were cold stored for 18 h, flushed with 20 ml of saline, kept at room temperature for 20 min, and reperfused at 37 °C with oxygenated Williams E solution. In half of the cases, the flush solution was supplemented with a cocktail containing metformin, bucladesine and cyclosporin A. Upon reperfusion, treated livers disclosed a massive mitigation of hepatic release of alanine aminotransferase and aspartate aminotransferase, along with a significant approximately 50 % reduction of radical mediated lipid peroxidation, caspase activation and release of TNF-alpha. Even after preceding cold preservation, a pharmacological cocktail given as single flush is capable to mitigate manifestations of reperfusion injury in the present model.


Asunto(s)
Ciclosporina , Peroxidación de Lípido , Hígado , Preservación de Órganos , Ratas Wistar , Daño por Reperfusión , Factor de Necrosis Tumoral alfa , Animales , Daño por Reperfusión/prevención & control , Daño por Reperfusión/tratamiento farmacológico , Masculino , Ratas , Hígado/efectos de los fármacos , Hígado/metabolismo , Hígado/irrigación sanguínea , Preservación de Órganos/métodos , Ciclosporina/farmacología , Peroxidación de Lípido/efectos de los fármacos , Factor de Necrosis Tumoral alfa/metabolismo , Metformina/farmacología , Metformina/uso terapéutico , Alanina Transaminasa/metabolismo , Alanina Transaminasa/sangre , Aspartato Aminotransferasas/metabolismo , Recalentamiento/métodos , Soluciones Preservantes de Órganos/farmacología
20.
BMC Microbiol ; 24(1): 189, 2024 May 30.
Artículo en Inglés | MEDLINE | ID: mdl-38811884

RESUMEN

BACKGROUND: The study aims to analyze the epidemiology of preservation fluid (PF) contamination and investigate the impact of PF contamination and possible donor-derived infections(p-DDI) on early postoperative prognosis in kidney transplant (KT) recipients. METHODS: A total of 256 PF samples were collected for microbiological evaluation from all KT recipients who received deceased donor donations in our hospital from June 2018 to August 2022. Data on the baseline and clinical characteristics of these PF corresponding to recipients and donors were extracted from the electronic medical record. It mainly included the early postoperative complications and prognosis of KT recipients. RESULTS: From June 2018 to August 2022, 597 kidney transplants were performed in our center, with 260 recipients receiving kidney transplantation from donation after citizens' death. A total of 256 samples of PF were collected, of which 64.5% (165/256) were culture positive, and 24.6% (63/165) of the culture-positive PF were polymicrobial contamination. A total of 238 strains were isolated, of which coagulase-negative staphylococci (CoNS) had the highest proportion of 34.0% (81/238), followed by Klebsiella pneumoniae with 20.6% (49/238) and Escherichia coli with 8.8% (21/238). Recipients with culture-positive PF had a significantly higher incidence of postoperative infection (55.8% vs. 20.9%, P < 0.001) and DGF (38.2% vs. 24.2%, P = 0.023). In addition, the incidence of p-DDI was 12.9% (33/256). CRKP was the most common pathogen causing p-DDI. The recipients who developed p-DDI had a higher rate of graft loss (9.1% vs. 0.4%, P < 0.001), mortality (12.1% vs. 3.1%, P = 0.018), and longer postoperative hospital stay (30 days (19.5-73.5) vs. (22 days (18-32), P < 0.05) compared with recipients who did not develop p-DDI. CONCLUSIONS: Culture-positive PF is potentially significant for KT recipients, and p-DDI may increase the risk of poor prognosis for recipients. Prophylactic anti-infective treatment should be actively performed for highly virulent or multidrug-resistant (MDR) pathogens (especially Carbapenem-resistant Klebsiella pneumoniae, CRKP) in PF to avoid the occurrence of p-DDI.


Asunto(s)
Trasplante de Riñón , Soluciones Preservantes de Órganos , Donantes de Tejidos , Humanos , Trasplante de Riñón/efectos adversos , Masculino , Femenino , Persona de Mediana Edad , Adulto , Pronóstico , Complicaciones Posoperatorias/microbiología , Complicaciones Posoperatorias/epidemiología , Receptores de Trasplantes/estadística & datos numéricos , Estudios Retrospectivos , Anciano , Bacterias/aislamiento & purificación , Bacterias/clasificación , Bacterias/genética
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