Air embolism during liver procurement: an underestimated phenomenon? A pilot experimental study.

BACKGROUND: Intrahepatic air embolism can occur during liver transplantation, jeopardizing the posttransplant outcome. Until now, the role of the procurement in the origin of intrahepatic air remains unclear; it might be underestimated. In this pilot study using magnetic resonance imaging (MRI), we observed a substantial amount of air trapped in porcine livers during multiorgan procurement. We quantified the amount of air, examining whether it could be reduced by avoiding direct contact of air with the lumen of the hepatic vasculature during procurement and back-table preparation. METHODS: Five livers (control group) were procured according to standard techniques for comparison with 6 livers (modified group) where air could not enter into the livers due to clamping of the vasculature. MRI was performed during continuous machine perfusion (MP) preservation there after. We counted the number of black signal voids on T(2)*-weighted images, which were indicative of air bubbles within the hepatic contour. Additionally, an MRI contrast agent (gadolinium-diethylene triamine pentaacetic acid [Gd-DTPA]) was injected into the hepatic artery and circulated by MP. Insufficiently perfused areas with less contrast enhancement were analyzed quantitatively in T(1)-weighted images and expressed as the percentage of total liver volume. RESULTS: The images of the control livers showed more air bubbles compared with the modified group (45 ± 27 vs 6 ± 3; P = .004). The percentage of insufficiently perfused areas was higher among the control compared with the modified group (28.0 ± 15.8% vs 2.6 ± 4.6%; P = .047) on first-pass postcontrast T(1)-weighted images. After recirculating the contrast agent, insufficiently perfused areas showed similar localizations and contours within every liver. CONCLUSION: These data suggested that a substantial amount of air enters into the hepatic microcirculation through direct contact of air with the hepatic vasculature during standard procurement and back-table preparation. Avoiding opening the hepatic vessels to air substantially reduced this phenomenon.

Discriminate liver warm ischemic injury during hypothermic machine perfusion by proton magnetic resonance spectroscopy: a study in a porcine model.

OBJECTIVE: Proton magnetic resonance spectroscopy ((1)H MRS) is a technique to identify and quantify the composition of biofluids. Hypothermic machine perfusion (HMP) is an organ preservation method that has the potential to evaluate organ viability prior to transplantation by analyzing the composition of the perfusate. The aim of this study was to use (1)H MRS to examine the perfusate during HMP of porcine livers exposed to warm ischemia (WI) and to identify potential biomarkers of liver viability. MATERIALS AND METHODS: Porcine livers underwent 4 hours of HMP using kidney perfusion solution-1 (KPS-1) as perfusate after exposure to in situ WI of 0 (n = 6) or 2 hours (n = 5). Samples of the perfusate were taken at the start/end of HMP. Lactate and aspartate aminotransferase (AST) in the samples were measured biochemically as surrogates of the WI-induced damage. MRS acquisition was conducted on a 9.4 Tesla (400 MHz) high-resolution system. RESULTS: AST increased significantly during 4 hours of HMP within groups (P < .02) and discriminated WI injury significantly from the start to the end of HMP (P < .03). (1)H MRS showed significantly elevated signal intensity of lactate, alanine, and histidine during HMP within both groups (P < .02). Furthermore, alanine and histidine were significantly higher in the WI group than in the control group at the end of HMP (P = .011 and P = .038, respectively). CONCLUSION: AST, alanine, and histidine in HMP perfusate discriminated WI injury on porcine liver grafts and might be potential biomarkers of liver viability.

Twenty-four hour hypothermic machine perfusion preservation of porcine pancreas facilitates processing for islet isolation.

Procurement of donor pancreata for islet isolation and transplantation is not yet widely practiced due to concerns about the impact of postmortem ischemia on functional islet yields. Perfusion/preservation technology may help to circumvent ischemic injury as applied in this study of porcine pancreata prior to islet isolation. Pancreata harvested from adult pigs were assigned to 1 of 3 preservation treatment groups: G1, fresh controls, processed immediately with minimum cold ischemia (<1 hour); G2, static cold storage, flushed with cold UW-Viaspan and stored at 2 degrees -4 degrees C for 24 hours; and G3, hypothermic machine perfusion (HMP) on a pulsatile LifePort machine Organ Recovery Systems, Inc., Des Plaines, Ill, United States with KPS1 solution at 4-7 degrees C and low pressure (10 mm Hg) for 24 hours. Islet isolation was then accomplished using conventional methods. Product release criteria were used to assess islet yield and function. Islet yield was markedly different between the treatment groups. There was a statistically significant increased yield in the HMP group over static cold storage in UW-Viaspan (P < .05). Functionally, the islets from each experimental group were equivalent and not significantly different from fresh controls (G1). Dithizone staining of islets showed consistently more uniform digestion of pancreata from G3 compared with G1 and G2, with greater separation of the tissue and fewer entrapped islets. HMP for 24 hours was well tolerated, leading to moderate edema but no loss of function of the harvested islets. The edema appeared to aid in enzymatic digestion, producing a greater yield and purity of islets compared with pancreata subjected to 24 hours of static cold storage.

Can apparent diffusion coefficient discriminate ischemic from nonischemic livers? A pilot experimental study.

PURPOSE: Using magnetic resonance imaging, the apparent diffusion coefficient (ADC) is an indicator to assess cerebral ischemia. The aim of this porcine study was to evaluate whether ADC assessed hepatic ischemia during ex vivo hypothermic machine perfusion (HMP) as well as in vivo. METHODS: Ex vivo: ADC of normal versus warm ischemic (WI) livers was assessed during HMP and subsequent rewarming to mimic ischemia-reperfusion injury. As the preservation solution, we used either an acellular solution or diluted blood. WI was induced in the left lobe or in the whole liver and compared 2-hour WI and non-WI. In vivo: One liver was scanned with the left lobe vessels occluded for 2-hour WI and subsequently for 3 hour reperfusion to compare with the right lobe without WI. Aspartate aminotransferase (AST) in the perfusate and morphology were used as surrogates of WI. RESULTS: In all WI livers, AST reached high levels and histology showed severe injury. Ex vivo ADC during acellular perfusion showed negligible differences between the livers with versus without WI, namely, 0.75 x 10(-3) or 0.88 x 10(-3) mm(2)/s during HMP. Ex vivo ADC using sanguineous perfusion showed 1.11 x 10(-3) or 0.83 x 10(-3) mm(2)/s during HMP in regions with versus without WI, respectively, a difference that remained stable during the whole experiment. ADC in vivo decreased from the physiological level of 1.07 x 10(-3) mm(2)/s to 0.75 x 10(-3) mm(2)/s in the first 30 minutes of WI, whereas ADC in the non-WI liver remained constant. CONCLUSION: ADC in vivo decreased during hepatic ischemia, as previously seen in cerebral ischemia. However, the effect of WI on ADC was less clear during ex vivo HMP.

Influence of flow and addition of oxygen during porcine liver hypothermic machine perfusion.

INTRODUCTION: In contrast with kidneys, data on hypothermic machine perfusion (HMP) of livers remain scarce. Optimal liver HMP is poorly defined. Superiority of liver HMP over simple cold storage (SCS), the current standard preservation, must be proven before HMP is applied clinically. In this study, morphology and adenosine triphosphate (ATP) contents of HMP livers at different flows and with versus without O(2) studied in a porcine ex vivo model were compared to SCS. METHODS: Pig livers were procured, flushed with HTK and preserved via SCS or HMP at 3 HMP settings: high flow (HF); low flow (LF); low flow + O(2) (300 mm Hg) (LFO). HMP livers were perfused via the hepatic artery (HA) and portal vein (PV) with KPS-1 TM at 4 degrees C to 6 degrees C for 24 hours with HF: PV: 3 to 5 mm Hg, 1 mL/g liver/min for HA and 25 mm Hg; LF: PV: 3 to 5 mm Hg, 0.5 ml/g liver/min with HA: 20 mm Hg. Morphology and ATP levels were measured in preserved liver tissues. RESULTS: Throughout the SCS preservation, livers remained intact. In HMP livers, vacuoles appeared after 4 hours of preservation in the HF group and after 12 hours in the LF livers. LFO livers remained intact with limited vacuoles. Compared to SCS, HMP livers showed dilated sinusoids, particularly in the HF group. ATP remained relatively constant or even increased during HMP, particularly in the LF group, whereas ATP decreased after SCS. CONCLUSION: Among the various HMP settings, HMP with LFO was superior. ATP levels were the highest in LF. In contrast with all HMP groups, SCS showed the lowest ATP levels, indicating that HMP has the potential to better preserve energy stores.

Hemodynamic, biochemical, and morphological characteristics during preservation of normal porcine livers by hypothermic machine perfusion.

In renal transplantation, hypothermic machine perfusion optimizes preservation of marginal grafts, assesses their quality prior to transplantation, improves outcome, and may contribute to an increased number of transplantations. Recently, hypothermic machine perfusion has become increasingly popular given the organ shortage and the "obligatory" utilization of marginal organs. Increasing mortality on the liver transplantation waiting list makes it urgent to develop machine perfusion systems for livers, trying to better preserve marginal livers and perhaps to recover currently discarded livers are for clinical transplantation without an increased risk of graft nonfunction. However, data on machine perfusion of livers and perfusion parameters capable of predicting viability are scarce. The aim of this study was to determine the baseline hemodynamic and metabolic profiles and morphology of livers during hypothermic machine perfusion in a porcine model. We used protocol similar to hypothermic machine perfusion of kidneys. Hemodynamic analysis revealed higher vascular resistance in the hepatic artery versus the portal vein. The arterial resistance gradually decreased during perfusion (similar to kidneys), suggesting progressive relaxation of the arterial vasculature, and perhaps better penetration of the microcirculation by the perfusion solution. During hypothermic machine perfusion, transaminases were gradually (but modestly) released, and livers displayed unequivocal signs of aerobic and anaerobic metabolism. After 24 hours, livers appeared morphologically well preserved. In conclusion, this study showed that hypothermic machine perfusion was feasible. During hypothermic machine perfusion, was easily assessed hemodynamic, biochemical, and morphological parameters.

Hepatic function in hypothermically stored porcine livers: comparison of hypothermic machine perfusion vs cold storage.

Hypothermic machine perfusion (HMP) has a potential to relieve the current donor liver crisis by providing an improved and extended preservation method. This study examined the effect of HMP on hepatocellular functions, using a prototype liver transporter capable of preserving livers for 24 hours. Livers obtained from adult farm pigs (28 to 32 kg body weight) were divided into three groups: fresh control, HMP, and simple cold storage (n = 4 each). A 4-hour normothermic reperfusion of livers was conducted to assess hepato-metabolic and cellular functions. The hepatic transport function, as indicated by canalicular excretion of indocyanine green, was improved in the HMP group than in the SCS group. The overall tissue viability, as indicated by oxygen consumption levels, was notably improved in HMP and control livers as compared to the SCS group. Higher bile production in both the preserved groups as compared to the fresh control livers could be a result of biliary edema and leakage of plasma into the canaliculus. The hepato-cellular injury, measured by ALT, release was significantly greater in the SCS group as compared to the HMP and control groups. These findings suggest that HMP could be a better method to preserve hepatic function and overall tissue viability as compared to SCS. Improved hepatic functions are indirect indicators of superior microcirculation and sinusoidal endothelial cell functions. Further studies in progress will evaluate these functions to confirm the significance of these observations.

Does ex vivo vascular resistance reflect viability of non-heart-beating donor livers?

AIMS: The use of non-heart-beating (NHB) donor livers is limited by a higher risk for primary nonfunction and the absence of methods to measure this risk. This study was designed to determine whether ex vivo vascular resistance of livers correlates with the length of warm ischemia (WI), and, thus, with viability of NHB livers. METHODS: Porcine livers were recovered after 0, 45, or 90 minutes WI. Livers were flushed by gravity and cold stored for 3 hours. Thereafter, livers were perfused at 4 degrees C. Portal vein (PV) and hepatic artery (HA) vascular resistance were calculated during liver flush-out and during 24 hours of machine perfusion. RESULTS: During flush-out, PV and HA vascular resistance were higher among livers with longer WI times; however, only in the PV did the results reach statistical significance. During machine perfusion, PV vascular resistance was low from the start and remained fairly constant. In contrast, HA vascular resistance was higher at the start but gradually diminished to reach a more constant value after 4-6 hours. No correlation was observed between HA or PV vascular resistance and WI during machine perfusion. CONCLUSIONS: The vascular resistance during ex vivo machine perfusion of NHB livers does not correlate with the extent of WI damage and, therefore, cannot predict organ viability.