Dominik Geisel

Porcine liver decellularization under oscillating pressure conditions: a technical refinement to improve the homogeneity of the decellularization process.

Decellularization and recellularization of parenchymal organs may facilitate the generation of autologous functional liver organoids by repopulation of decellularized porcine liver matrices with induced liver cells. We present an accelerated (7 h overall perfusion time) and effective protocol for human-scale liver decellularization by pressure-controlled perfusion with 1% Triton X-100 and 1% sodium dodecyl sulfate via the hepatic artery (120 mmHg) and portal vein (60 mmHg). In addition, we analyzed the effect of oscillating pressure conditions on pig liver decellularization (n=19). The proprietary perfusion device used to generate these pressure conditions mimics intra-abdominal conditions during respiration to optimize microperfusion within livers and thus optimize the homogeneity of the decellularization process. The efficiency of perfusion decellularization was analyzed by macroscopic observation, histological staining (hematoxylin and eosin [H&E], Sirius red, and alcian blue), immunohistochemical staining (collagen IV, laminin, and fibronectin), and biochemical assessment (DNA, collagen, and glycosaminoglycans) of decellularized liver matrices. The integrity of the extracellular matrix (ECM) postdecellularization was visualized by corrosion casting and three-dimensional computed tomography scanning. We found that livers perfused under oscillating pressure conditions (P(+)) showed a more homogenous course of decellularization and contained less DNA compared with livers perfused without oscillating pressure conditions (P(-)). Microscopically, livers from the (P(-)) group showed remnant cell clusters, while no cells were found in livers from the (P(+)) group. The grade of disruption of the ECM was higher in livers from the (P(-)) group, although the perfusion rates and pressure did not significantly differ. Immunohistochemical staining revealed that important matrix components were still present after decellularization. Corrosion casting showed an intact vascular (portal vein and hepatic artery) and biliary framework. In summary, the presented protocol for pig liver decellularization is quick (7 h) and effective. The application of oscillating pressure conditions improves the homogeneity of perfusion and thus the outcome of the decellularization process.

DNA double-strand breaks as potential indicators for the biological effects of ionising radiation exposure from cardiac CT and conventional coronary angiography: a randomised, controlled study

AbstractObjectivesTo prospectively compare induced DNA double-strand breaks by cardiac computed tomography (CT) and conventional coronary angiography (CCA).Methods56 patients with suspected coronary artery disease were randomised to undergo either CCA or cardiac CT. DNA double-strand breaks were assessed in fluorescence microscopy of blood lymphocytes as indicators of the biological effects of radiation exposure. Radiation doses were estimated using dose–length product (DLP) and dose–area product (DAP) with conversion factors for CT and CCA, respectively.ResultsOn average there were 0.12 ± 0.06 induced double-strand breaks per lymphocyte for CT and 0.29 ± 0.18 for diagnostic CCA (P < 0.001). This relative biological effect of ionising radiation from CCA was 1.9 times higher (P < 0.001) than the effective dose estimated by conversion factors would have suggested. The correlation between the biological effects and the estimated radiation doses was excellent for CT (r = 0.951, P < 0.001) and moderate to good for CCA (r = 0.862, P < 0.001). One day after radiation, a complete repair of double-strand breaks to background levels was found in both groups.ConclusionsConversion factors may underestimate the relative biological effects of ionising radiation from CCA. DNA double-strand break assessment may provide a strategy for individualised assessments of radiation.Key Points• Radiation dose causes concern for both conventional coronary angiography and cardiac CT. • Estimations of the biological effects of ionising radiation may become feasible. • Fewer DNA double-strand breaks are induced by cardiac CT than CCA. • Conversion factors may underestimate the relative effects of ionising radiation from CCA.

Is There Long-term Signal Intensity Increase in the Central Nervous System on T1-weighted Images after MR Imaging with the Hepatospecific Contrast Agent Gadoxetic Acid? A Cross-sectional Study in 91 Patients

Purpose To evaluate whether there is T1-weighted signal intensity (SI) increase in the dentate nucleus (DN) and globus pallidus (GP) in relation to the middle cerebellar peduncle (MCP), pons, and thalamus after repeated administration of the liver-specific contrast agent gadoxetic acid. Materials and Methods This was an institutional review board-approved, prospectively conducted (written informed consent acquired), cross-sectional study performed in a consecutively selected patient group (n = 91; patients received one to 37 doses of gadoxetic acid) and a control group (n = 52; subjects had never received injections of gadolinium-based contrast agent) examined with a standard T1-weighted two-dimensional spin-echo pulse sequence of the brain at 1.5 T. DN/MCP, DN-to-pons, GP-to thalamus, and GP-to-cerebrospinal fluid ratios were measured and compared by using the nonparametric Kruskal-Wallis test, corresponding pairwise tests, and Spearman correlation. Results DN/MCP (ρ = 0.51, P < .0001) and DN-to-pons (ρ = 0.41, P = .0001) ratios correlated positively with the number of previous administrations of gadoxetic acid. DN/MCP and DN-to-pons ratios were significantly different between control subjects (medians of 1.016 and 1.034, respectively) and patients with more than 10 gadoxetic acid administrations (1.038 [P < .0001] and 1.053 [P = .0100], respectively), whereas no significant difference was found in the groups with five to 10 (1.029 [P = .053] and 1.044 [P = .072], respectively) and fewer than five (1.014 [P = .420] and 1.030 [P = .595], respectively) gadoxetic acid administrations. GP-to-thalamus ratios differed significantly between the study and control groups (P < .0001), whereas no significant correlation was found for GP-to-thalamus ratios and number of gadoxetic acid administrations (ρ = 0.13, P = .2304). Conclusion Results show a significant correlation between the number of gadoxetic acid administrations and the increase of SI in the DN, which is likely due to gadolinium retention.

Portal vein embolization with plug/coils improves hepatectomy outcome

BACKGROUND Portal vein embolization (PVE) has become the standard of care before extended hepatectomy. Various PVE methods using different embolization materials have been described. In this study, we compared PVE with polyvinyl alcohol particles alone (PVA only) versus PVA with plug or coils (PVA + plug/coils). MATERIALS AND METHODS Patients undergoing PVE before hepatectomy were included. PVA alone was used until December 2013, thereafter plug or coils were placed in addition. The volume of left lateral liver lobe (LLL), clinical parameters, and liver function tests were measured before PVE and resection. RESULTS A total of 43 patients were recruited into the PVA only group and 42 were recruited into the PVA + plug/coils group. There were no major differences between groups except significantly higher total bilirubin level before PVE in the PVA only group, which improved before hepatectomy. Mean LLL volume increased by 25.7% after PVE in the PVA only group and by 44% in the PVA + plug/coils group (P < 0.001). Recanalization was significantly less common in the PVA + plug/coils group. In multivariate regression, initial LLL volume and use of plug or coils were the only parameters influencing LLL volume increase. The postoperative liver failure rate was significantly reduced in PVA + plug/coils group (P = <0.001). CONCLUSIONS PVE using PVA particles together with plug or coils is a safe and efficient method to increase future liver remnant volume. The additional central embolization with plug or coils led to an increased hypertrophy, due to lower recanalization rates, and subsequently decreased incidence of postoperative liver failure. No additional procedure-specific complications were observed in this series.

Improved rat liver decellularization by arterial perfusion under oscillating pressure conditions

One approach of regenerative medicine to generate functional hepatic tissue in vitro is decellularization and recellularization, and several protocols for the decellularization of livers of different species have been published. This appears to be the first report on rat liver decellularization by perfusion under oscillating pressure conditions, intending to optimize microperfusion and minimize damage to the ECM. Four decellularization protocols were compared: perfusion via the portal vein (PV) or the hepatic artery (HA), with (+P) or without (–P) oscillating pressure conditions. All rat livers (n = 24) were perfused with 1% Triton X‐100 and 1% sodium dodecyl sulphate, each for 90 min with a perfusion rate of 5 ml/min. Perfusion decellularization was observed macroscopically and the decellularized liver matrices were analysed by histology and biochemical analyses (e.g. levels of DNA, glycosaminoglycans and hepatocyte growth factor). Livers decellularized via the hepatic artery and under oscillating pressure showed a more homogeneous decellularization and less remaining DNA, compared with the livers of the other experimental groups. The novel decellularization method described is effective, quick (3 h) and gentle to the extracellular matrix and thus represents an improvement of existing methodology. Copyright

Imaging-Based Liver Function Tests--Past, Present and Future.

UNLABELLED Preoperative assessment of liver function and prediction of postoperative functional reserve are important in patients scheduled for liver resection. While determination of absolute liver function currently mostly relies on laboratory tests and clinical scores, postoperative remnant liver function is estimated volumetrically using imaging data obtained with computed tomography (CT) or magnetic resonance imaging (MRI). Accurate estimation of hepatic function is also relevant for intensive care patients, oncologic patients, and patients with diffuse liver disease. The indocyanine green (ICG) test is still the only established test for estimating true global liver function. However, more recent tools such as the LiMAx test also allow global assessment of hepatic function. These tests are limited when liver function is inhomogeneously distributed, which is the case in such conditions as unilateral cholestasis or after portal vein embolization. Imaging-based liver function tests were first developed in nuclear medicine and, compared with laboratory tests, have the advantage of displaying the spatial distribution of liver function. Nuclear medicine scans are obtained using tracers such as 99mTc galactosyl and 99mTc mebrofenin. Liver function is typically assessed using planar scintigraphy. However, three-dimensional volumetry is possible with single-photon emission computed tomography (SPECT-CT). Another technique for image-based liver function estimation is Gd-EOB-enhanced MRI. While metabolization of Gd-EOB in the body is similar to that of ICG and mebrofenin, its distribution in the liver can be displayed by MRI with higher temporal and spatial resolution. Moreover, MRI-based determination of liver function can be integrated into routine preoperative imaging. This makes MRI an ideal candidate for preoperative determination of liver function, though the best pulse sequence and the parameter to be derived from the image information remain to be identified. Another question to be answered is how the results may be affected by renal function and the presence of hyperbilirubinemia. As more results from clinical evaluation including comparison with postoperative liver function data become available, image-based liver function tests, especially with use of Gd-EOB as the contrast medium, have the potential to add another dimension to preoperative imaging. KEY POINTS Liver function consists of a multitude of subfunctions such as biotransformation, excretion and storage. Global liver function tests are score-based tests such as Child-Pugh or MELD as well as the ICG- and LiMAx-test. Imaging-based liver function tests add spatial information. Current clinical standard is the 99mTc-Mebrofenin-scintigraphy. MRI-based function tests with Gd-EOB-DTPA have the potential to integrate seamlessly into clinical workup, feature a higher temporal and spatial resolution and do not rely on ionizing radiation.

Hepatopulmonary shunting in patients with primary and secondary liver tumors scheduled for radioembolization

PURPOSE In patients undergoing transarterial radioembolization (RE) of malignant liver tumors, hepatopulmonary shunts (HPS) can lead to nontarget irradiation of the lungs. This study aims at analyzing the HPS fraction in relation to liver volume, tumor volume, tumor-to-liver volume ratio, tumor vascularity, type of tumor, and portal vein occlusion. MATERIALS AND METHODS In the presented retrospective study the percentage HPS fraction was calculated from SPECT/CT after infusion of Tc-99m macroaggregated albumin (Tc-99m MAA) into the proper hepatic artery of 233 patients evaluated for RE. RESULTS HPS fractions correlate very weakly with liver volume (r=0.303), tumor volume (r=0.345), and tumor-to-liver volume ratio (r=0.340). Tumors with strong contrast enhancement (HPSmedian(range)=11.7%(46.3%); n=73) have significantly larger shunt fractions than tumors with little enhancement (HPS=8.3%(16.4%); n=61; p<0.001). Colorectal cancer metastases (HPS=10.6%(28.6%); n=68) and hepatocellular cancers (HPS=11.7%(39.4%); n=63) have significantly larger HPS fractions than metastases from breast cancer (HPS=7.4%(16.7%); n=40; p=0.012 and p=0.001). Patients with compression (HPS=13.9%(43.7%); n=33) or tumor thrombosis (HPS=15.8% (31.2%); n=33) of a major portal vein branch have significantly higher degrees of shunting than patients with normal portal vein perfusion (HPS=8.1% (47.0%); n=167; both p<0.001). The shunt fraction is largest in patients with HCC and thrombosis or occlusion of a major portal vein branch (HPS=16.6% (31.0%); n=32). CONCLUSION The degree of hepatopulmonary shunting depends on the type of liver tumor, tumor vascularity, and portal vein perfusion. There is little to no correlation of HPS with liver volume, tumor volume, or tumor-to-liver volume ratio.

Evaluation of gadolinium-EOB-DTPA uptake after portal vein embolization: value of an increased flip angle.

Background The optimal sequence for Gd-EOB-DTPA uptake measurement in the liver with the purpose of liver function measurement is still not defined. Purpose To prospectively evaluate the effect of an increased flip angle (FA) of a T1-weighted fat-saturated 3D sequence for the measurement of hepatocyte uptake of Gd-EOB-DTPA magnetic resonance imaging (MRI) after right portal vein embolization (PVE). Material and Methods Ten patients who received a PVE prior to an extended hemihepatectomy were examined 14 days after PVE using Gd-EOB-DTPA enhanced MRI of the liver using the standard FA of 10° and the increased FA of 30°. Results Relative enhancement of the right liver lobe (RLL) was 0.52 ± 0.12 for 10° and 1.41 ± 0.39 for 30°. Relative enhancement of the left liver lobe (LLL) was 0.58 ± 0.11 for 10° and 2.05 ± 0.61 for 30°. Relative enhancement of the RLL was significantly higher for 30° than for 10° (P = 0.009) and significantly higher in the 30° than in the 10° sequences (P = 0.005) for the LLL. Conclusion A flip angle of 30° increases the contrast between liver partitions with and without portal venous embolization. Thereby, the sensitivity for differences in uptake intensity is increased. This could be of value for a more exact determination of differences in regional liver function and, consequently, the estimation of the future remnant liver function.

Factors influencing hypertrophy of the left lateral liver lobe after portal vein embolization

PurposePortal vein embolization (PVE) before extended right hepatectomy leads to an increase of the future liver remnant (FLR) volume, but predictive factors for sufficient hypertrophy are still unclear. The purpose of this study was to investigate parameters influencing the growth of FLR.MethodsPatients undergoing PVE prior hepatic resection were evaluated. PVE was done using polyvinyl alcohol particles only. Volumetric analysis was performed before embolization and before hepatectomy. Success of PVE was determined as percental growth of the future liver remnant.ResultsSeventy-seven patients were included, and three cohorts were formed according to the hypertrophy of FLR. FLR increased from 448.2 ± 187 to 475.5 ± 191 in the poor, from 315.3 ± 86 to 469.1 ± 142 in the moderate, and from 283.4 ± 68 to 400.4 ± 110 in the good hypertrophy group. More cases of recanalization of the portal vein were observed in patients with poor hypertrophy (p = 0.016). Small FLR before PVE predict higher growth of the FLR (p = 0.006). Duration between PVE and surgery differed significantly: 22 (poor) vs. 32 (good) days (p = 0.040).DiscussionNo recanalization, small initial FLR and longer time were assessed with better FLR hypertrophy. More sufficient PVE techniques and postponed hepatectomy might improve the outcome. Small initial FLR should not be a disclosure for curative hepatectomy.

Anatomic variants of arteries often coil-occluded prior to hepatic radioembolization

Background: Prior to radioembolization (RE) treatment of malignant liver lesions, many interventionalists occlude the right gastric artery (RGA), the cystic artery (CA), and the gastroduodenal artery (GDA) to prevent radioactive microspheres from entering non-target vessels. Purpose: To systematically analyze anatomic variants of arteries that are important to know for the interventional radiologist performing RE of the liver. Material and Methods: The computed tomography (CT) angiographies and conventional angiographies of 166 patients evaluated for RE were retrospectively analyzed for the presence of anatomic variants of the RGA, GDA, and CA. Results: The RGA was found to arise from the left hepatic artery in 42% of cases, from the proper hepatic artery in 40%, from the GDA in 10%, from the right hepatic artery in 4%, and from the common hepatic artery in 3% of cases. The GDA originated in the common hepatic artery in 97% of cases, in the left hepatic artery in 2%, and in the celiac trunk in 1% of cases. The CA arose from the right hepatic artery in 96% of cases and from the GDA in 2% of cases; in 2% of our study population, the gallbladder was supplied by small branches from the liver parenchyma. Conclusion: Variant anatomy of the RGA is common, while it is quite rare for the GDA and CA. Knowledge of the variations of liver supplying arteries helps the interventionalist to embolize necessary vessels prior to RE.