TY - GEN A1 - Birkenmaier, Clemens A1 - Krenkel, Lars A1 - Lehle, Karla T1 - Linking flow conditions in membrane oxygenators to arrangements of multimeric von-Willebrand-factor as indication for coagulation T2 - World Congress of Biomechanics 2018, Convention Centre Dublin, 8.-12. Juli 2018 N2 - Introduction Shear induced multimerisation of von-Willebrand-factor (vWF) is supposed to play an important role in coagulation inside extracorporeal membrane oxygenators. However, there is no proof that links observed vWF structures to computed or measured flow conditions. Methods The structures of multimeric vWF fibers, observed in clinically used membrane oxygenators is examined using immunofluorescence microscopy (IFM) using Carstairs’ staining method (positive ethics committee vote). The flow around the membrane fibres inside the oxygenator is investigated in terms of shear rate, wall shear velocity and streamlines by using CFD (RANS, Carreau-Yasuda viscosity, geometry remodelled after high-resolution µCT-scans). By interpreting the histological and numerical results in this common context, indications for shear induced coagulation mechanisms can be identified. Results The fibre structures of multimeric vWF build regular but not exactly symmetric formations around the contact face (CF) between the crosswise stacked oxygenator fibres (OF), see fig.1B, vWF marked red. Annular around the CF arranged, cells are likely to be found, see fig.1B, nuclei marked blue. The computed streamlines around the OF show attached flow around the circular fibres. However, the irregular arrangement of real OF produce considerable cross flow between the interconnected neighbouring channels, in contrast to previous 2D-simulations. Thus, the CF are washed around closely by blood, also from neighbouring channels. The wall shear velocity streamlines form regular, slightly asymmetric shapes around the contact faces. The occurring maximum shear rates are in the range of 1,000 1/s. Discussion The shapes of vWF structures found in clinically used oxygenators match the computational results in terms of wall shear velocity and streamlines well. The accumulation of cells close to the CF can also be explained by fluid mechanics, as there are small shear gradients and slow velocities. However, occurring shear rates between OFs are too low to trigger multimerisation of vWF. That raises the question where in the circuit the actual activation of vWF is started and how, at least partly chained, vWF multimeres are attracted towards the OF surface. A next step will be the investigation of the actual shear rate triggered (or mediated) multimerisation of vWF. Towards this end, microfluidic experiments with shear triggered coagulation will be performed. Also of big interest is the computation of the flow situation in the oxygenator in proximity to chaining threads, which have been ignored in computations so far. However, first a realistic representation of the effective viscosity in computations is needed, which is not available yet. Y1 - 2018 ER - TY - JOUR A1 - Philipp, Alois A1 - de Somer, Filip A1 - Foltan, Maik A1 - Bredthauer, Andre A1 - Krenkel, Lars A1 - Zeman, Florian A1 - Lehle, Karla T1 - Life span of different extracorporeal membrane systems for severe respiratory failure in the clinical practice JF - PLOS ONE N2 - Over the past decade, veno-venous extracorporeal membrane oxygenation (vvECMO) has been increasingly utilized in respiratory failure in patients. This study presents our institution´s experience focusing on the life span of ECMO systems reflecting the performance of a particular system. A retrospective review of our ECMO database identified 461 adult patients undergoing vvECMO (2010-2017). Patients that required more than one system and survived the first exchange >24 hours (n = 139) were included. Life span until the first exchange and exchange criteria were analyzed for all systems (PLS, Cardiohelp HLS-set, both Maquet Cardiopulmonary, Rastatt, Germany; Deltastream/Hilite7000LT, iLA-activve, Xenios/NovaLung, Heilbronn, Germany; ECC.O5, LivaNova, Mirandola, Italy). At our ECMO center, the frequency of a system exchange was 30%. The median (IQR) life span was 9 (6-12) days. There was no difference regarding the different systems (p = 0.145 and p = 0.108, respectively). However, the Deltastream systems were exchanged more frequently due to elective technical complications (e. g. worsened gas transfer, development of coagulation disorder, increased bleedings complications) compared to the other exchanged systems (p = 0.013). In summary, the used ECMO systems are safe and effective for acute respiratory failure. There is no evidence for the usage of a specific system. Only the increased predictability of an imminent exchange preferred the usage of a Deltastream system. However, the decision to use a particular system should not depend solely on the possible criteria for an exchange. KW - Equipment Failure Analysis/statistics & numerical data KW - Extracorporeal Membrane Oxygenation/instrumentation KW - Membrane/classification/standards/statistics & numerical data KW - Primary Health Care/statistics & numerical data KW - Respiratory Distress Syndrome/therapy KW - Retrospective Studies KW - Severity of Illness Index KW - Time factors KW - MULTIDETECTOR COMPUTED-TOMOGRAPHY KW - THROMBOTIC DEPOSITS KW - ECMO SYSTEMS KW - Flow KW - OXYGENATION Y1 - 2018 U6 - https://doi.org/10.1371/journal.pone.0198392 VL - 13 IS - 6 SP - 1 EP - 10 PB - PLOS ER - TY - RPRT A1 - Steiger, Tamara A1 - Foltan, Maik A1 - Philipp, Alois A1 - Müller, Thomas A1 - Gruber, Michael A1 - Bredthauer, Andre A1 - Krenkel, Lars A1 - Birkenmaier, Clemens A1 - Lehle, Karla T1 - Accumulations of von Willebrand factor within ECMO oxygenators: Potential indicator of coagulation abnormalities in critically ill patients? N2 - Clot formation within membrane oxygenators (MOs) remains a critical problem during extracorporeal membrane oxygenation (ECMO). The composition of the clots—in particular, the presence of von Willebrand factor (vWF)—may be an indicator for prevalent nonphysiological flow conditions, foreign body reactions, or coagulation abnormalities in critically ill patients. Mats of interwoven gas exchange fibers from randomly collected MOs (PLS, Maquet, Rastatt, Germany) of 21 patients were stained with antibodies (anti‐vWF and anti‐P‐selectin) and counterstained with 4′,6‐diamidino‐2‐phenylindole. The extent of vWF‐loading was correlated with patient and technical data. While 12 MOs showed low vWF‐loadings, 9 MOs showed high vWF‐loading with highest accumulations close to crossing points of adjacent gas fibers. The presence and the extent of vWF‐fibers/“cobwebs,” leukocytes, platelet–leukocyte aggregates (PLAs), and P‐selectin‐positive platelet aggregates were independent of the extent of vWF‐loading. However, the highly loaded MOs were obtained from patients with a significantly elevated SOFA score, severe thrombocytopenia, and persistent liver dysfunction. The coagulation abnormalities of these critically ill patients may cause an accumulation of the highly thrombogenic and elongated high‐molecular‐weight vWF multimers in the plasma which will be trapped in the MOs during the ECMO therapy. Y1 - 2019 ER - TY - CHAP A1 - Birkenmaier, Clemens A1 - Steiger, Tamara A1 - Philipp, Alois A1 - Lehle, Karla A1 - Krenkel, Lars T1 - Flow-induced accumulations of von Willebrand factor inside oxygenators during extracorporeal life support therapy T2 - Proceedings of 12th International Conference BIOMDLORE 2018, June 28–30, 2018, Białystok, Poland N2 - BACKGROUND: Shear-induced conformational changes of von Willebrand factor (vWF) may be responsible for coagulation disorder and clot formation inside membrane oxygenators (MOs) during extracorporeal membrane oxygenation (ECMO) therapy. OBJECTIVE: The aim was to identify vWF structures inside clinically used MOs and employ computational fluid dynamics to verify the corresponding flow conditions. METHODS: Samples from gas exchange membranes (GEM) from MOs were analysed for accumulations of vWF and P-selectin-positive platelets using immunofluorescence techniques. Streamlines and shear rates of the flow around GEMs were computed using a laminar steady Reynolds-Averaged-Navier-Stokes approach. RESULTS: Most samples were colonized with equally distributed leukocytes, integrated in thin cobweb-like vWF-structures. Only 25 % of the samples showed extended accumulations of vWF. Computed streamlines showed considerable cross flow between interconnected neighbouring channels. Stagnation points were non-symmetric and contact faces were washed around closely. The occurring maximum shear rates ranged from 2,500 to 3,000 1/s. CONCLUSIONS: If pronounced vWF structures are present, shape and extent match the flow computations well. Computed shear rates bear a critical degree of uncertainty due to the improper viscosity model. If flow conditions inside the MO were sufficient to affect vWF, a more consistent distribution of vWF across the samples should be present. KW - Blood Viscosity KW - Shear Rate Induced Coagulation KW - Hemodynamics KW - Membrane Oxygenator KW - von Willebrand factor Y1 - 2018 SN - 978-1-5386-2396-1 U6 - https://doi.org/10.1109/BIOMDLORE.2018.8467205 PB - IEEE CY - Piscataway, NJ ER - TY - JOUR A1 - Birkenmaier, Clemens A1 - Dornia, Christian A1 - Lehle, Karla A1 - Müller, Thomas A1 - Gruber, Michael A1 - Philipp, Alois A1 - Krenkel, Lars T1 - Analysis of Thrombotic Deposits in Extracorporeal Membrane Oxygenators by High-resolution Microcomputed Tomography: A Feasibility Study JF - ASAIO Journal / American Society for Artificial Internal Organs N2 - Coagulative disorders, especially clotting during extracorporeal membrane oxygenation, are frequent complications. Direct visualization and analysis of deposits in membrane oxygenators using computed tomography (CT) may provide an insight into the underlying mechanisms causing thrombotic events. However, the already established multidetector CT1 (MDCT) method shows major limitations. Here, we demonstrate the feasibility of applying industrial micro-CT (μCT) to circumvent these restrictions. Three clinically used membrane oxygenators were investigated applying both MDCT and μCT. The scans were analyzed in terms of clot volume and local clot distribution. As validation, the clot volume was also determined from the fluid volume, which could be filled into the respective used oxygenator compared to a new device. In addition, cross-sectional CT images were compared with crosscut oxygenators. Based on the μCT findings, a morphological measure (sphericity) for assessing clot structures in membrane oxygenators is introduced. Furthermore, by comparing MDCT and μCT results, an augmentation of the MDCT method is proposed, which allows for improved clot volume determination in a clinical setting. Y1 - 2020 U6 - https://doi.org/10.1097/MAT.0000000000001089 SN - 1538-943X VL - 66 IS - 8 SP - 922 EP - 928 PB - Lippincott Williams & Wilkins ER - TY - GEN A1 - Birkenmaier, Clemens A1 - Dornia, Christian A1 - Lehle, Karla A1 - Krenkel, Lars T1 - Feasibility of detecting thrombotic deposits in membrane oxygenators using micro computed tomography T2 - 25th Congress of the European Society of Biomechanics, July 7-10, 2019, Vienna, Austria Y1 - 2019 UR - https://esbiomech.org/conference/archive/2019vienna/Contribution_129.pdf ER - TY - JOUR A1 - Obermaier, Lisa A1 - Lehle, Karla A1 - Schmid, Stefanie A1 - Schmid, Christof A1 - Schratzenstaller, Thomas T1 - Introduction of a new ex vivo porcine coronary artery model: Evaluation of the direct vascular injury after stent implantation with and without dogbone effect JF - European Surgical Research N2 - Introduction: Neointimal hyperplasia after percutaneous coronary intervention remains a major determinant of in-stent restenosis (ISR). The extent of mechanical vessel injury correlates with ISR. A new ex vivo porcine stent model was introduced and evaluated comparing different stent designs. Methods: Coronary arteries were prepared from pig hearts from the slaughterhouse and used for ex vivo implantations of coronary stents. One basic stent design in two configurations (dogbone, DB; non-dogbone, NDB) was used. Vascular injury was determined according to a modified injury score (IS). Results: Standardized experimental conditions ensured comparable vessel dimensions and overstretch data. DB stents caused more severe IS compared to NDB stents. The mean IS and the IS at the distal end of all stents were significantly reduced for NDB stents (ISMean, DB, 1.16 ±0.12; NDB, 1.02 ±0.12; p=0.018; ISDist, DB, 1.39 ±0.28; NDB, 1.13 ±0.24; p=0.03). Discussion/Conclusion: The introduced ex-vivo model allowed the evaluation of different stent designs exclude unfavorable stent designs. KW - Stent screening KW - Stent design KW - Injury score KW - Ex vivo porcine stent model Y1 - 2022 U6 - https://doi.org/10.1159/000527883 SN - 1421-9921 VL - 63 IS - 4 SP - 285 EP - 293 PB - Karger CY - Basel ER - TY - INPR A1 - Thaus, Christopher A1 - Hofrichter, Elena A1 - Lubnow, Matthias A1 - Krenkel, Lars A1 - Lehle, Karla T1 - Hemocompatibility of Membrane Lung Components from Extracorporeal Membrane Oxygenation with Different Antithrombogenic Coatings N2 - Thrombus formation within extracorporeal membrane oxygenation (ECMO) devices remained a critical complication. One reason seems to be the contact of blood with large artificial surfaces within the membrane lung (ML). The aim was to test the hemocompatibility of different naïve ECMO materials. Blood and platelets from five healthy volunteers were incubated with gas exchange (GF) and heat exchange fibers (HE) from four different commercial available new MLs representing different antithrombogenic coatings. Adherent platelets were stained with rhodamine-phalloidin. Surface coverage was quantified with ImageJ. Non-adherent platelets were stained with antibodies (CD62P, PAC-1, CD61) and fibrinogen to detect platelet activation with flow cytometry. Hemolysis of red blood cells after material contact was detected. All ECMO-materials were non-hemolytic and did not induce platelet activation. However, platetelet adhesion (median (IQR)) was significantly elevated on uncoated GFs made of polymethylpentene (GF-PMP; 12 (7-19)%) and on GFs from the Hilite-MLs (GF-Hilite; 13 ((8-19)%) compared to the other materials. In vitro testing of platelet adhesion disclosed significant differences of ECMO-materials with different antithrombogenic surface coatings. Instead, circulating platelets remained non-activated. ECMO-materials and its coatings were non-hemolytic. Finally, this study confirmed the good hemocompatibility of GFs and HEs from commerciall available MLs. KW - ECMO KW - platelet activation KW - platelet adhesion KW - hemolysis KW - hemocompatibility Y1 - 2024 U6 - https://doi.org/10.20944/preprints202412.0615.v1 SP - 1 EP - 11 ER - TY - JOUR A1 - Hoenicka, Markus A1 - Lehle, Karla A1 - Jacobs, V. R. A1 - Dendorfer, Sebastian A1 - Kostorz, A. A1 - Schmid, F. X. A1 - Birnbaum, D. E. T1 - Mechanical and seeding properties of human umbilical vein – a potential scaffold for a tissue-engineered vessel graft JF - The Thoracic and Cardiovascular Surgeon N2 - Objectives: The mechanical properties and seeding with endothelial cells were investigated in fresh and cryopreserved human umbilical vein. Methods: Human umbilical veins (HUV) were frozen in Euro-Collins/1M DMSO at –1°C/min and stored in liquid nitrogen. Stress-strain relationships of fresh and thawed veins were determined in an uniaxial tension-testing rig. HUV endothelial cells (HUVEC) were seeded onto denuded HUV under static conditions and grown for 3d. Luminal surfaces were analyzed by scanning electron microscopy. Calcein-stained cells were seeded hyperconfluently to determine the cell retention capacity of fresh and cryopreserved veins. Results: The stress-strain relationships of HUV followed a biphasic pattern typical for natural vessels. Neither the failure stress (2.71±0.36 vs. 3.25±0.97 N, n=3) nor the displacement required to achieve failure (9.73±0.9 vs. 7.43±2.07mm, n=3) were altered by cryopreservation. The burst pressure was estimated as approx. 1000mm Hg within the limitations of the uniaxial model. HUVEC seeded onto denuded HUV formed patches (at 9E3 cells per cm2) or an almost confluent endothelium (at 3E4 cells per cm2) within three days. The capacity to retain seeded HUVEC of denuded HUV was not altered by cryopreservation (1.15±0.08E5 vs. 1.26±0.14E5 cells per cm2, n=6). Conclusions: The burst pressure of HUV seems to be sufficiently high for the human arterial circulation and is not altered by cryopreservation. HUVEC can establish a confluent endothelium on denuded HUV. Therefore HUV appears to be a suitable storable scaffold for vascular tissue engineering. KW - Nabelvene KW - Tissue Engineering Y1 - 2007 U6 - https://doi.org/10.1055/s-2007-967592 VL - 55 IS - S 1 SP - P_37 PB - Thieme ER - TY - JOUR A1 - Wagner, Maria Stella A1 - Kranz, Michael A1 - Krenkel, Lars A1 - Pointner, Daniel A1 - Foltan, Maik A1 - Lubnow, Matthias A1 - Lehle, Karla ED - Becatti, Matteo T1 - Computer based visualization of clot structures in extracorporeal membrane oxygenation and histological clot investigations for understanding thrombosis in membrane lungs JF - Frontiers in Medicine N2 - Extracorporeal membrane oxygenation (ECMO) was established as a treatment for severe cardiac or respiratory disease. Intra-device clot formation is a common risk. This is based on complex coagulation phenomena which are not yet sufficiently understood. The objective was the development and validation of a methodology to capture the key properties of clots deposed in membrane lungs (MLs), such as clot size, distribution, burden, and composition. One end-oftherapy PLS ML was examined. Clot detection was performed using multidetector computed tomography (MDCT), microcomputed tomography (μCT), and photography of fiber mats (fiber mat imaging, FMI). Histological staining was conducted for von Willebrand factor (vWF), platelets (CD42b, CD62P), fibrin, and nucleated cells (4′, 6-diamidino-2-phenylindole, DAPI). The three imaging methods showed similar clot distribution inside the ML. Independent of the imaging method, clot loading was detected predominantly in the inlet chamber of the ML. The μCT had the highest accuracy. However, it was more expensive and time consuming than MDCT or FMI. The MDCT detected the clots with low scanning time. Due to its lower resolution, it only showed clotted areas but not the exact shape of clot structures. FMI represented the simplest variant, requiring little effort and resources. FMI allowed clot localization and calculation of clot volume. Histological evaluation indicated omnipresent immunological deposits throughout the ML. Visually clot-free areas were covered with leukocytes and platelets forming platelet-leukocyte aggregates (PLAs). Cells were embedded in vWF cobwebs, while vWF fibers were negligible. In conclusion, the presented methodology allowed adequate clot identification and histological classification of possible thrombosis markers such as PLAs. KW - ECMO KW - membrane lung KW - µCT KW - MDCT KW - shear induced clotting KW - vWF KW - histological evaluation Y1 - 2024 U6 - https://doi.org/10.3389/fmed.2024.1416319 IS - 11 PB - Frontiers ER - TY - JOUR A1 - Lingel, Maximilian P. A1 - Haus, Moritz A1 - Paschke, Lukas A1 - Foltan, Maik A1 - Lubnow, Matthias A1 - Gruber, Michael A1 - Krenkel, Lars A1 - Lehle, Karla T1 - Clinical relevance of cell-free DNA during venovenous extracorporeal membrane oxygenation JF - Artificial organs N2 - BACKGROUND: Thrombosis remains a critical complication during venovenous extracorporeal membrane oxygenation (VV ECMO). The involvement of neutrophil extracellular traps (NETs) in thrombogenesis has to be discussed. The aim was to verify NETs in the form of cell-free DNA (cfDNA) in the plasma of patients during ECMO. METHODS: A fluorescent DNA-binding dye (QuantifFluor®, Promega) was used to detect cell-free DNA in plasma samples. cfDNA concentrations from volunteers (n = 21) and patients (n = 9) were compared and correlated with clinical/technical data before/during support, ECMO end and time of a system exchange. RESULTS: Before ECMO, patients with a median (IQR) age of 59 (51/63) years, SOFA score of 11 (10/15), and ECMO run time of 9.0 (7.0/19.5) days presented significantly higher levels of cfDNA compared to volunteers (6.4 (5.8/7.9) ng/μL vs. 5.9 (5.4/6.3) ng/μL; p = 0.044). Within 2 days after ECMO start, cfDNA, inflammatory, and hemolysis parameters remained unchanged, while platelets decreased (p = 0.005). After ECMO removal at the end of therapy, cfDNA, inflammation, and coagulation data (except antithrombin III) remained unchanged. The renewal of a system resulted in known alterations in fibrinogen, d-dimers, and platelets, while cfDNA remained unchanged. CONCLUSION: Detection of cfDNA in plasma of ECMO patients was not an indicator of acute and circuit-induced thrombogenesis. KW - blood KW - cell- free DNA KW - coagulation KW - ECMO KW - inflammation KW - neutrophil extracellular traps Y1 - 2023 U6 - https://doi.org/10.1111/aor.14616 SN - 1525-1594 VL - 47 IS - 11 SP - 1720 EP - 1731 PB - Wiley ER - TY - JOUR A1 - Foltan, Maik A1 - Dinh, D. A1 - Gruber, Michael A1 - Müller, Thomas A1 - Hart, C. A1 - Krenkel, Lars A1 - Schmid, C. A1 - Lehle, Karla T1 - Incidence of neutrophil extracellular traps (NETs) in different membrane oxygenators: pilot in vitro experiments in commercially available coated membranes JF - Journal of artificial organs : the official journal of the Japanese Society for Artificial Organs N2 - Neutrophil extracellular traps (NETs) were detected in blood samples and in cellular deposits of oxygenator membranes during extracorporeal membrane oxygenation (ECMO) therapy and may be responsible for thrombogenesis. The aim was to evaluate the effect of the base material of gas fiber (GF, polymethylpentene) and heat exchange (HE) membranes and different antithrombogenic coatings on isolated granulocytes from healthy volunteers under static culture conditions. Contact of granulocytes with membranes from different ECMO oxygenators (with different surface coatings) and uncoated-GFs allowed detection of adherent cells and NETotic nuclear structures (normal, swollen, ruptured) using nuclear staining. Flow cytometry was used to identify cell activation (CD11b/CD62L, oxidative burst) of non-adherent cells. Uncoated-GFs were used as a reference. Within 3 h, granulocytes adhered to the same extent on all surfaces. In contrast, the ratio of normal to NETotic cells was significantly higher for uncoated-GFs (56-83%) compared to all coated GFs (34-72%) (p < 0.001) with no difference between the coatings. After material contact, non-adherent cells remained vital with unchanged oxidative burst function and the proportion of activated cells remained low. The expression of activation markers was independent of the origin of the GF material. In conclusion, the polymethylpentene surfaces of the GFs already induce NET formation. Antithrombogenic coatings can already reduce the proportion of NETotic nuclei. However, it cannot be ruled out that NET formation can induce thrombotic events. Therefore, new surfaces or coatings are required for future ECMO systems and long-term implantable artificial lungs. Y1 - 2025 U6 - https://doi.org/10.1007/s10047-024-01486-4 ER - TY - JOUR A1 - Paschke, Lukas A1 - Foltan, Maik A1 - Wagner, Maria S. A1 - Lubnow, Matthias A1 - Gruber, Michael A1 - Krenkel, Lars A1 - Lehle, Karla T1 - Clinical Relevance of Platelet-Leukocyte Aggregates and Platelet P-Selectin Expression During Venovenous Extracorporeal Membrane Oxygenation JF - ASAIO Journal N2 - Thrombosis continues to be a significant complication during venovenous extracorporeal membrane oxygenation (V-V ECMO). Platelet activation markers might serve as indicators of inflammation and thrombogenesis. The aim was to identify these markers in ECMO patients. Blood from 10 ECMO patients (before, during, after ECMO) and 11 healthy volunteers were collected to determine platelet-neutrophil-aggregates (PNAs), platelet-monocyte-aggregates (PMAs), fibrinogen-binding, and P-selectin-expression on platelets by flow cytometry. Critical illness was associated with significantly elevated levels of PNAs and PMAs, increased P-selectin expression, reduced fibrinogen-binding, and restricted activation of platelets. Although PNAs and PMAs decreased significantly within 2 hours after the initiation of ECMO and remained at those levels, ECMO did not affect basal P-selectin expression and fibrinogen-binding. These results correlated with coagulation activation. Platelet markers before ECMO were not indicators for an imminent system exchange and end of therapy. In conclusion, platelet dysfunction during ECMO was mainly attributed to the critical illness. Extracorporeal membrane oxygenation support strengthened the restricted response of platelets to exogenous agonists (P-selectin). Furthermore, a decrease in PNAs/PMAs after ECMO started identified a reduced inflammatory response. There was no correlation of analyzed platelet parameters with the incidence of thrombotic complications. Y1 - 2025 U6 - https://doi.org/10.1097/MAT.0000000000002421 SN - 1058-2916 IS - April 03 PB - Wolters Kluwer ER - TY - JOUR A1 - Steiger, Tamara A1 - Foltan, Maik A1 - Philipp, Alois A1 - Müller, Thomas A1 - Gruber, Michael A1 - Bredthauer, Andre A1 - Krenkel, Lars A1 - Birkenmaier, Clemens A1 - Lehle, Karla T1 - Accumulations of von Willebrand factor within ECMO oxygenators: Potential indicator of coagulation abnormalities in critically ill patients? JF - Artificial Organs N2 - Clot formation within membrane oxygenators (MOs) remains a critical problem during extracorporeal membrane oxygenation (ECMO). The composition of the clots-in particular, the presence of von Willebrand factor (vWF)-may be an indicator for prevalent nonphysiological flow conditions, foreign body reactions, or coagulation abnormalities in critically ill patients. Mats of interwoven gas exchange fibers from randomly collected MOs (PLS, Maquet, Rastatt, Germany) of 21 patients were stained with antibodies (anti-vWF and anti-P-selectin) and counterstained with 4 ',6-diamidino-2-phenylindole. The extent of vWF-loading was correlated with patient and technical data. While 12 MOs showed low vWF-loadings, 9 MOs showed high vWF-loading with highest accumulations close to crossing points of adjacent gas fibers. The presence and the extent of vWF-fibers/"cobwebs," leukocytes, platelet-leukocyte aggregates (PLAs), and P-selectin-positive platelet aggregates were independent of the extent of vWF-loading. However, the highly loaded MOs were obtained from patients with a significantly elevated SOFA score, severe thrombocytopenia, and persistent liver dysfunction. The coagulation abnormalities of these critically ill patients may cause an accumulation of the highly thrombogenic and elongated high-molecular-weight vWF multimers in the plasma which will be trapped in the MOs during the ECMO therapy. KW - ECMO KW - PLATELET ACTIVATION KW - THROMBOSIS KW - BLOOD FLOW KW - INFLAMMATION Y1 - 2019 U6 - https://doi.org/10.1111/aor.13513 SN - 1525-1594 VL - 43 IS - 11 SP - 1065 EP - 1076 PB - Wiley CY - Hoboken ER - TY - JOUR A1 - Haus, Moritz A1 - Foltan, Maik A1 - Philipp, Alois A1 - Müller, Thomas A1 - Lingel, Maximilian P. A1 - Krenkel, Lars A1 - Gruber, Michael A1 - Lehle, Karla T1 - Neutrophil extracellular traps -a potential trigger for the development of thrombocytopenia during extracorporeal membrane oxygenation N2 - Neutrophil extracellular traps (NETs) have recently emerged as a potential link between inflammation, immunity, and thrombosis, as well as other coagulation disorders which present a major challenge in the context of extracorporeal membrane oxygenation (ECMO). By examining blood from ECMO patients for NETs and their precursors and correlating them with clinical and laboratory biomarkers of coagulation and inflammation, this study aims to evaluate the association between the presence of NETs in the bloodstream of ECMO patients and the development of potentially severe coagulation disorders during ECMO therapy. Therefore, blood samples were collected from healthy volunteers (n=13) and patients receiving veno-venous (VV) ECMO therapy (n=10). To identify NETs and their precursors, DNA and myeloperoxidase as well as granulocyte marker CD66b were visualized simultaneously by immunofluorescence staining in serial blood smears. Differentiation of DNA-containing objects and identification of NETs and their precursors was performed semiautomatically by a specific algorithm using the shape and size of DNA staining and the intensity of MPO and CD66b signal. Neutrophil extracellular traps and their precursors could be detected in blood smears from patients requiring VV ECMO. Compared to volunteers, ECMO patients presented significantly higher rates of NETs and NET precursors as well as an increased proportion of neutrophil granulocytes in all detected nucleated cells. A high NET rate prior to the initiation of ECMO therapy was associated with both increased iL-6 and TNF-α levels as an expression of a high cytokine burden. These patients with increased NET release also presented an earlier and significantly more pronounced decrease in platelet counts and ATIII activity following initiation of therapy compared with patients with less elevated NETs. These findings provide further indications for the development of immune-mediated acquired thrombocytopenia in ECMO patients. KW - Neutrophil extracellular traps (NET) KW - Immunoflorescence KW - Thrombocytopenia KW - Extracorporeal membrane oxygenation (ECMO) KW - Sepsis KW - immunothrombosis KW - Coagulation disorder Y1 - 2024 UR - https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2024.1339235/abstract U6 - https://doi.org/10.3389/fimmu.2024.1339235 VL - 15 PB - frontiers ER - TY - JOUR A1 - Lehle, Karla A1 - Philipp, Alois A1 - Krenkel, Lars A1 - Gruber, Michael A1 - Hiller, Karl-Anton A1 - Müller, Thomas A1 - Lubnow, Matthias T1 - Thrombocytopenia During Venovenous Extracorporeal Membrane Oxygenation in Adult Patients With Bacterial, Viral, and COVID-19 Pneumonia JF - ASAIO Journal N2 - Contact of blood with artificial surfaces triggers platelet activation. The aim was to compare platelet kinetics after venovenous extracorporeal membrane oxygenation (V-V ECMO) start and after system exchange in different etiologies of acute lung failure. Platelet counts and coagulation parameters were analyzed from adult patients with long and exchange-free (≥8 days) ECMO runs (n = 330) caused by bacterial (n = 142), viral (n = 76), or coronavirus disease 2019 (COVID-19) (n = 112) pneumonia. A subpopulation requiring a system exchange and with long, exchange-free runs of the second oxygenator (≥7 days) (n = 110) was analyzed analogously. Patients with COVID-19 showed the highest platelet levels before ECMO implantation. Independent of the underlying disease and ECMO type, platelet counts decreased significantly within 24 hours and reached a steady state after 5 days. In the subpopulation, at the day of a system exchange, platelet counts were lower compared with ECMO start, but without differences between underlying diseases. Subsequently, platelets remained unchanged in the bacterial pneumonia group, but increased in the COVID-19 and viral pneumonia groups within 2–4 days, whereas D-dimers decreased and fibrinogen levels increased. Thus, overall platelet counts on V-V ECMO show disease-specific initial dynamics followed by an ongoing consumption by the ECMO device, which is not boosted by new artificial surfaces after a system exchange. Y1 - 2025 U6 - https://doi.org/10.1097/MAT.0000000000002383 SN - 1058-2916 SN - 1538-943X PB - Wolters Kluwer ER - TY - JOUR A1 - Kranz, Michael A1 - Wagner, Maria Stella A1 - Pointner, Daniel A1 - Haus, Moritz A1 - Lubnow, Matthias A1 - Lehle, Karla A1 - Krenkel, Lars T1 - Polymer embedding of membrane lungs for histological investigations of intra-device clot formation JF - Cardiovascular Medicine N2 - Extracorporeal membrane oxygenation (ECMO) is an invasive but potentially lifesaving treatment option for severe cardiac or respiratory failure. Despite its beneficial effect, coagulation-related complications, mainly due to clot formation, excessive bleeding and the accumulation of deposits in the membrane lung (ML) remain common, causing higher mortality. In this context, the formation of clots and other deposits in the ML is of particular interest. Previous histological examinations of the polymethylpentene fiber mats inside the ML could only be performed in a top view, prohibiting valid quantification and examination of the multi-layered deposits or fiber mat spanning structures. Our objective was the establishment of a polymer embedding to increase the mechanical stability of the deposits and thus enable cross-sectional microtome cutting through the ML hollow-fibers. Clinically used MLs (PLS, Getinge, Rastatt, Germany) were stabilized with a polymer resin (HistoCURE 8100). Specimens were cut out of the embedded MLs and microtome sections with a thickness of 10 µm were performed. In addition to standard histological staining with hematoxylin-eosin (HE) and Pappenheim (May-Grunwald-Giemsa), fluorescence DNA staining for nucleated cells with 4′,6-diamidino-2-phenylindole (DAPI) and SYTOX™ Green as well as immunohistochemical and immunofluorescence staining for the lysosomal enzyme myeloperoxidase (MPO) and von Willebrand factor (vWF) were established. The protocol provides a method for large volume embedding (400 mL). The cellular and extracellular deposits were securely fixed by the polymer scaffold allowing the examination of clots in MLs in native position which was not possible with conventional paraffin embedding. Multi-layered deposits and fiber mat spanning structures are no longer disrupted during specimen extraction and can now be quantified. Staining with HE, Pappenheim, DAPI, SYTOX™ Green, MPO, and vWF was successfully tested with this protocol. This method may be the foundation for new insights into the complex clotting phenomena observed in MLs KW - clot formation KW - ECMO KW - HistoCURE 8100 KW - histology KW - membrane lungs KW - polymer embedding KW - Technovit 8100 Y1 - 2026 U6 - https://doi.org/10.3389/fcvm.2026.1650978 VL - 13 PB - Frontiers CY - Lausanne ER - TY - CHAP A1 - Kranz, Michael A1 - Pointner, Daniel A1 - Wagner, Maria Stella A1 - Lubnow, Matthias A1 - Lehle, Karla A1 - Krenkel, Lars ED - Dillmann, Andreas ED - Heller, Gerd ED - Krämer, Ewald ED - Breitsamter, Christian ED - Wagner, Claus ED - Krenkel, Lars T1 - High-Resolution Flow Investigations in Membrane-Lungs for Understanding Shear-Induced Blood Clot Formation T2 - New Results in Numerical and Experimental Fluid Mechanics XV : Contributions to the 24th STAB/DGLR Symposium, Regensburg, Germany, 2024 N2 - Complex blood flow phenomena in membrane lungs (MLs) play a crucial role in intra-device clot formation and the occurrence of thromboembolic events. At present, however, the local flow conditions within an ML are not yet sufficiently known. The aim was to gain a deeper understanding of local flow regimes inside MLs by performing highly resolved computational fluid dynamics (CFD) of generic and native fiber mat bundles. Straight cylinders with a diameter of 380 μm in parallel arrangement were the foundation of the generic model. For validation, a method for reconstructing a native geometry from a microcomputed tomography (μCT) scan was established, with both models used for CFD. While the generic model showed a symmetrical flow regime without indicating any pathological flow, the native model did show an irregular fiber arrangement and no symmetrical flow regime. In conclusion, the fiber arrangement significantly affects the local flow regimes inside MLs. KW - CFD KW - ECMO KW - membrane lung KW - ML KW - µCT Y1 - 2026 U6 - https://doi.org/10.1007/978-3-032-11115-9_12 SP - 125 EP - 134 PB - Springer CY - Cham ER - TY - GEN A1 - Kranz, Michael A1 - Pointner, Daniel A1 - Lehle, Karla A1 - Lubnow, Matthias A1 - Krenkel, Lars T1 - High-resolution flow field investigations in membrane lungs, considering the complex blood rheology T2 - 1st European Fluid Dynamics Conference (EFDC1), 16-20.September 2024, Aachen N2 - Despite major improvements over the last years, coagulative disorders and clotting phenomena in membrane lungs (MLs) are still considerable complications in extracorporeal membrane oxygenation (ECMO). ECMO is an increasingly used treatment for patients with severe respiratory failure or cardiac arrest [1]. For both, evaluation of therapeutic decisions and fundamental research on patient specific intra-device clotting phenomena, the direct visualization and analysis of clot formation in combination with a detailed flow field correlation is highly desirable and therefore an intensively followed research topic. Modelling blood flow and shear induced coagulation in MLs is challenging. The relevant geometry of oxygenator fibers and chaining threads is complex and spans several length scales. In relevant scales and regimes, blood shows several significant non-Newtonian effects. Viscosity impacts shear rate, which is important in several coagulation mechanisms. Additionally, coagulation processes are influencing fluid properties and geometry significantly. Existing approaches of previous research work are only able to consider some, but not all relevant effects and geometrical details. Due to the enormous size of the discretized geometries, highly detailed viscosity and coagulations models are not applicable. Our goal is to develop a model for combined viscosity and coagulation properties of blood flow in MLs. In our work, we compare the influence of different levels of detail of the ML geometry as well as the influence of considering realistic blood flow behavior (viscosity change by considering the local hematocrit distribution within the Fåhraeus-Lindqvist-Effect) on the resulting flow field in relevant subsections of a ML. High-resolution micro-CT geometry reconstructions [1] are compared to idealized generic fiber representations. For realistic blood flow modelling, Newtonian representation is compared to the established Carreau-Yasuda and a multiphase Euler-Euler approach. Results are presented for relevant subsections as well as for the complete ML. Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:898-opus4-89214 ER - TY - GEN A1 - Kranz, Michael A1 - Wagner, Maria Stella A1 - Pointner, Daniel A1 - Waldbauer, Selina A1 - Müller, Thomas A1 - Lubnow, Matthias A1 - Foltan, Maik A1 - Krenkel, Lars A1 - Lehle, Karla T1 - Polymeric Embedding of Membrane Lungs: A Novel Method for Histological Investigations of Intra-Device Clot Formation T2 - 12th EuroELSO Congress, 24-27. April 2024, Krakow Y1 - 2024 ER - TY - GEN A1 - Kranz, Michael A1 - Wagner, Maria Stella A1 - Krenkel, Lars A1 - Müller, Thomas A1 - Lubnow, Matthias A1 - Philipp, Alois A1 - Lehle, Karla T1 - Clot Localization within Membrane Lungs using different Imaging Methods and Histological Clot Characterization as a way to prevent Thrombosis in Extracorporeal Membrane Oxygenation Y1 - 2023 VL - 2023 ER - TY - GEN A1 - Köster, Leonie A1 - Kranz, Michael A1 - Wagner, Maria Stella A1 - Foltan, Maik A1 - Müller, Thomas A1 - Lubnow, Matthias A1 - Krenkel, Lars A1 - Lehle, Karla T1 - Histological Investigations of Intra-Device Clot Formation in ECMO Pumps T2 - 12th EuroELSO Congress, 24-27. April 2024, Krakow Y1 - 2024 ER - TY - JOUR A1 - Pointner, Daniel A1 - Kranz, Michael A1 - Wagner, Maria Stella A1 - Haus, Moritz A1 - Lehle, Karla A1 - Krenkel, Lars T1 - Automated deep learning based detection of cellular deposits on clinically used ECMO membrane lungs JF - Frontiers in Bioinformatics N2 - Introduction: Despite the promising application of extracorporeal membrane oxygenation (ECMO) in the treatment of critically ill patients, coagulation-associated technical complications, primarily clot formation and critical bleeding, remain a major challenge during ECMO therapy. The deposition of nucleated cells on the surface has been shown, yet the role of these cells towards complication development is still matter of ongoing research. In particular, the membrane lung (MemL) is prone to clot formation. Therefore, the investigation of nuclear deposits on its hollow-fibers may provide insights for a better understanding of the cellular mechanisms involved in the development of ECMO complications. Methods: To support current research, this study aimed to develop a deep learning–based tool for the automated detection and quantitative analysis of nuclear depositions on MemL hollow-fiber mats. A customized fluorescence microscopy workflow, combined with a semi-automated iterative labeling strategy, was used to generate a high-quality dataset for model training. Results: Six configurations of instance segmentation models were evaluated, with a Mask R-CNN with ResNet 101 backbone using dilated convolution providing the most balanced performance in both nuclei count and area accuracy. Compared with U-Net–based approaches such as Cellpose or StarDist, the proposed model demonstrated superior segmentation of overlapping and low-intensity nuclei, maintaining accuracy even in densely packed cellular regions. Discussion: We present an automated image analysis tool for clinically used MemLs, which exhibit complex three-dimensional hollow-fiber architectures and irregular cellular deposits that challenge conventional tools. A dedicated graphical user interface enables streamlined detection, morphometric analysis, and spatial clustering of nuclei, establishing a reproducible workflow for high-throughput analysis of fluorescence microscopy images. This approach eliminates labor-intensive manual counting and facilitates large-scale studies on cell-fiber interactions and disease-related correlations. Y1 - 2026 U6 - https://doi.org/10.3389/fbinf.2026.1771574 N1 - Corresponding author der OTH Regensburg: Daniel Pointner, Lars Krenkel VL - 6 PB - Frontiers ER -