Influence on energy kinetics and histology of different preservation solutions seen during cold ischemia in the liver.

Transplant Science Group, Department of Hepatology and Transplantation, St. James's University Hospital, Leeds West Yorkshire LS9 7TF, United Kingdom.
Transplantation Proceedings (Impact Factor: 0.95). 12/2009; 41(10):4088-93. DOI: 10.1016/j.transproceed.2009.07.107
Source: PubMed

ABSTRACT Cold flush preservation prolongs tissue viability during ischemia. However, there is little understanding of the effects of various preservation fluids on events during this period. A study of cold ischemia in rat livers was undertaken to compare biochemical and histological changes over time, using three preservation solutions: University of Wisconsin (UW), histidine-tryptophan-ketoglutarate (HTK), and Leeds solution (LS) under development at our institution. Leeds solution is a phosphate-based sucrose solution that like UW contains the impermeant lactobionate and the metabolite allopurinol (1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one) which acts as a competitive inhibitor of xanthine oxidase, stopping the breakdown of hypoxanthine to xanthine by oxidizing it to alloxanthine, inhibiting both the conversion of hypoxanthine to xanthine and the conversion of xanthine to uric acid.
At various time points, samples were analyzed for adenosine triphospate (ATP) and metabolites by high-performance liquid chromatography as well as for histological changes.
In all livers, ATP, ADP, and AMP degraded over 4 hours. In UW and LS groups, degradation beyond hypoxanthine was halted, and it continued in the HTK group. This blockade led to a significant reduction in the accumulation of xanthine and uric acid. Histological analysis showed protected architecture and maintenance of reticulin scaffolds in the UW and LS groups, whereas tissue breakdown was seen from earlier time points in the HTK group. Additionally, throughout ischemia, signs of pathological injury were more pronounced with UW- than with LS-preserved tissue.
These results implied that cold ischemia in the liver is characterized by dynamic biochemical changes coincident with pathological injury which are initiated from the time of organ perfusion and influenced by the choice of the perfusion fluid. Allopurinol in UW and LS appears to be critical. We hypothesized that it may also affect the degree of subsequent reperfusion injury. The data supported the assertion that LS offerred improved preservation over UW, adding to the impetus to shorten ischemic times in clinical transplantation.

  • [Show abstract] [Hide abstract]
    ABSTRACT: In order to gain further insight into the mechanisms of tissue damage during the progression of liver diseases as well as the liver preservation for transplantation, an improved understanding of the relation between the mechanical and histological properties of liver is necessary. We suggest that this relation can only be established truly if the changes in the states of those properties are investigated dynamically as a function of post mortem time. In this regard, we first perform mechanical characterization experiments on three bovine livers to investigate the changes in gross mechanical properties (stiffness, viscosity, and fracture toughness) for the preservation periods of 5, 11, 17, 29, 41 and 53h after harvesting. Then, the histological examination is performed on the samples taken from the same livers to investigate the changes in apoptotic cell count, collagen accumulation, sinusoidal dilatation, and glycogen deposition as a function of the same preservation periods. Finally, the correlation between the mechanical and histological properties is investigated via the Spearman's Rank-Order Correlation method. The results of our study show that stiffness, viscosity, and fracture toughness of bovine liver increase as the preservation period is increased. These macroscopic changes are very strongly correlated with the increase in collagen accumulation and decrease in deposited glycogen level at the microscopic level. Also, we observe that the largest changes in mechanical and histological properties occur after the first 11-17h of preservation.
    Journal of the Mechanical Behavior of Biomedical Materials 01/2014; 29:403-416. · 2.37 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Sediment–turbulence interactions near the bed are still poorly understood in highly turbid estuaries, especially in the presence of fluid mud layers. This results primarily from the difficulty in measuring co-located velocity and suspended sediment concentration (SSC) at sufficiently high rate to resolve small turbulent flow scales. In this paper, we show how a set of commercial acoustic and optical backscattering systems known as ADCPs, ADVs and OBSs, can be deployed and used in a complementary way to perform large-scale profilings of tidal current and SSC combined with high-resolution velocity and SSC measurements in the highly turbid near-bed zone. The experiment was done in the Gironde estuary (France) which is well known for its turbidity maximum zone characterized by high SSC values, above 1 g l−1 near the surface.A first simple inversion method is proposed to convert the backscattered acoustic intensity measured with ADV into SSC data in the highly turbid near-bed zone. Near-bed SSC data from the OBS are used to compensate for the important acoustic sediment attenuation effect at an acoustic frequency of 6 MHz. No a priori knowledge of acoustic backscattering properties of mud suspensions is required with this calibration procedure. We obtain an attenuation coefficient for mud suspensions of 0.28 m2/kg at 6 MHz leading to a good agreement between the SSC timeseries from the three ADV receivers and the OBS over the entire tidal cycle.The obtained SSC data are then analyzed with respect to the near-bed velocity, Reynolds shear stress and turbulent kinetic energy (TKE) timeseries in order to identify the relevant sediment transport processes during the tidal cycle. Significant differences in bed shear stress and TKE levels are found between ebb and flood stages with effects on near-bed sediment dynamics. During the ebb, maximum levels of tidal current, bed shear stress and TKE are associated with a reduction of near-bed sediment concentration (from 400 kg m−3 down to 100 kg m−3). Bed liquefaction process is assumed to occur at this moment with the presence of highly concentrated mud layer and a possible lutocline at a distance of less than 20 cm above the bed. During the first 1.5 h of flood, turbulent activity remains moderate. The near-bed flood current is then inhibited very abruptly while a sudden increase in SSC occurred above the bed.Assuming that the ADV is able to estimate relevant turbulent erosion fluxes, the co-located velocity and SSC are multiplied and compared with settling flux measurements made onboard under quiescent water conditions. The mean sediment settling fluxes (averaged over 3 min) increase with SSC and are in relative good coherence with fluxes in quiescent water below the hindered regime, for SSC below 15 g l−1. Reducing averaging time from 3 min to 30 s allows to increase the range of turbulent fluxes and SSC values, up to 99 g l−1. At this scale, fluxes keep increasing quasi-linearly at higher SSC, suggesting the inhibition (delay or reduction) of the hindered settling regime as previously shown by Gratiot et al. (2005) from laboratory experiments. However, the 3-min averaged concentration field remained too low to conclude definitively on the effectiveness of such a process. Further analysis conducted at higher SSC regimes and under fully verified equilibrium are necessary.Highlights► Near-bed turbulent fluxes were measured in the Gironde estuary by acoustic velocimetry. ► Acoustic velocities, turbulent shear and SSC were analyzed in the presence of fluid mud. ► Settling fluxes in quiescent waters and turbulent fluxes are compared in a wide range of SSC. ► Processes governing near-bed sediment fluxes under the effect of turbulence are discussed.
    Continental Shelf Research 07/2011; 31(10). · 2.12 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Introduction The solution in which graft tissue is stored (that is, preservation solution) is an important component of liver transplantation technology. Its protective effect is induced by substances in the solution, including radical scavengers, buffers, and energy-giving substances. New preservation solutions have proven to be effective in preventing organ damage during cold ischemia and in extending the time limits for storage. Aim This study determined the relationship between luminescence intensity and content of adenosine triphosphate (ATP) in liver tissue and proposes a new ex vivo screening system that uses Lewis rats transgenic for luciferase for evaluating the effectiveness of preservation solutions. Methods Samples (diameter, 2 mm) of liver were obtained from transgenic rats. The viability of these tissues after storage for as long as 6 hours in University of Wisconsin (UW) solution, extracellular trehalose solution of Kyoto, Euro-Collins (EC) solution, histidine–tryptophan–ketoflutarate solution, low potassium dextran solution, or normal saline was assessed by determining ATP content and luminescence intensity. Results Luminescence had a linear relationship (R = 0.88) with ATP levels. Regardless of the preservation solution used, the luminescence intensities of the liver tissue chips decreased linearly with time especially through a short span of time (0 to 2 hours; R2 = 0.58–1.0). The luminescence of liver chip tissues maintained long term (2 to 6 hours) in UW solution tended to be higher than those of tissues stored in other solutions (P < .05; 6 hours). On the basis of luminescence intensity, EC might be preferable to the other solutions tested for ultra-short–term storage (0.5 to 2 hours). Conclusion Our model, which combines the use of the bioimaging system and Lewis rats transgenic for luciferase, effectively assessed the viability of liver tissue samples. We believe that this ex vivo screening system will be an effective tool for evaluating preservation solutions for liver grafts.
    Transplantation Proceedings 01/2014; 46(1):63–65. · 0.95 Impact Factor