@inproceedings{MichelKrenkel, author = {Michel, Johanna and Krenkel, Lars}, title = {Towards Experimental Validation of Models of Shear-Induced Aerosol Generation in the Human Respiratory System}, series = {New Results in Numerical and Experimental Fluid Mechanics XV : Contributions to the 24th STAB/DGLR Symposium, Regensburg, Germany, 2024}, booktitle = {New Results in Numerical and Experimental Fluid Mechanics XV : Contributions to the 24th STAB/DGLR Symposium, Regensburg, Germany, 2024}, editor = {Dillmann, Andreas and Heller, Gerd and Kr{\"a}mer, Ewald and Breitsamter, Christian and Wagner, Claus and Krenkel, Lars}, publisher = {Springer Nature}, address = {Cham}, isbn = {978-3-032-11114-2}, doi = {10.1007/978-3-032-11115-9_13}, pages = {135 -- 144}, abstract = {Numerical modeling is a valuable tool to research shear-induced aerosol generation inside the human respiratory system. While the volume of fluid method and Eulerian wall film models have been used to predict the stripping of particles from the mucus film, sufficient validation data is lacking. Here, we present an experimental method to create such validation data. A film of mucus mimetic hydrogel with an initial thickness of 1 mm covering the floor of a rectangular channel (75.5 mm 25.5 mm 3 mm) was exposed to an airflow with a flow rate of 9.5 and 21.6 L/min. The number of created particles and the emergence of waves on the mucus surface were measured. Shear-induced aerosol generation was triggered successfully and caused an increase of mean particle flow. Different wave profiles were observed at varying film depths.}, language = {en} } @article{PointnerKranzWagneretal., author = {Pointner, Daniel and Kranz, Michael and Wagner, Maria Stella and Haus, Moritz and Lehle, Karla and Krenkel, Lars}, title = {Automated deep learning based detection of cellular deposits on clinically used ECMO membrane lungs}, series = {Frontiers in Bioinformatics}, volume = {6}, journal = {Frontiers in Bioinformatics}, publisher = {Frontiers}, doi = {10.3389/fbinf.2026.1771574}, pages = {18}, abstract = {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.}, language = {en} } @misc{KranzPointnerLehleetal., author = {Kranz, Michael and Pointner, Daniel and Lehle, Karla and Lubnow, Matthias and Krenkel, Lars}, title = {High-resolution flow field investigations in membrane lungs, considering the complex blood rheology}, series = {1st European Fluid Dynamics Conference (EFDC1), 16-20.September 2024, Aachen}, journal = {1st European Fluid Dynamics Conference (EFDC1), 16-20.September 2024, Aachen}, doi = {10.35096/othr/pub-8921}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:898-opus4-89214}, pages = {2}, abstract = {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 F{\aa}hraeus-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.}, language = {en} } @misc{KranzWagnerPointneretal., author = {Kranz, Michael and Wagner, Maria Stella and Pointner, Daniel and Waldbauer, Selina and M{\"u}ller, Thomas and Lubnow, Matthias and Foltan, Maik and Krenkel, Lars and Lehle, Karla}, title = {Polymeric Embedding of Membrane Lungs: A Novel Method for Histological Investigations of Intra-Device Clot Formation}, series = {12th EuroELSO Congress, 24-27. April 2024, Krakow}, journal = {12th EuroELSO Congress, 24-27. April 2024, Krakow}, language = {en} } @misc{KoesterKranzWagneretal., author = {K{\"o}ster, Leonie and Kranz, Michael and Wagner, Maria Stella and Foltan, Maik and M{\"u}ller, Thomas and Lubnow, Matthias and Krenkel, Lars and Lehle, Karla}, title = {Histological Investigations of Intra-Device Clot Formation in ECMO Pumps}, series = {12th EuroELSO Congress, 24-27. April 2024, Krakow}, journal = {12th EuroELSO Congress, 24-27. April 2024, Krakow}, language = {en} } @misc{KranzWagnerKrenkeletal., author = {Kranz, Michael and Wagner, Maria Stella and Krenkel, Lars and M{\"u}ller, Thomas and Lubnow, Matthias and Philipp, Alois and Lehle, Karla}, title = {Clot Localization within Membrane Lungs using different Imaging Methods and Histological Clot Characterization as a way to prevent Thrombosis in Extracorporeal Membrane Oxygenation}, volume = {2023}, language = {en} } @inproceedings{KranzPointnerWagneretal., author = {Kranz, Michael and Pointner, Daniel and Wagner, Maria Stella and Lubnow, Matthias and Lehle, Karla and Krenkel, Lars}, title = {High-Resolution Flow Investigations in Membrane-Lungs for Understanding Shear-Induced Blood Clot Formation}, series = {New Results in Numerical and Experimental Fluid Mechanics XV : Contributions to the 24th STAB/DGLR Symposium, Regensburg, Germany, 2024}, booktitle = {New Results in Numerical and Experimental Fluid Mechanics XV : Contributions to the 24th STAB/DGLR Symposium, Regensburg, Germany, 2024}, editor = {Dillmann, Andreas and Heller, Gerd and Kr{\"a}mer, Ewald and Breitsamter, Christian and Wagner, Claus and Krenkel, Lars}, publisher = {Springer}, address = {Cham}, doi = {10.1007/978-3-032-11115-9_12}, pages = {125 -- 134}, abstract = {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.}, language = {en} } @misc{MichelKrenkel, author = {Michel, Johanna and Krenkel, Lars}, title = {Experimental Investigation of Shear-Induced Generation of Respiratory Aerosol: Simultaneous Measurements of Particle Quantities and Wave Topology}, series = {2nd European Fluid Dynamics Conference (EFDC2), 26-29 August 2025, Dublin, Ireland}, journal = {2nd European Fluid Dynamics Conference (EFDC2), 26-29 August 2025, Dublin, Ireland}, doi = {10.35096/othr/pub-8859}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:898-opus4-88593}, pages = {1}, abstract = {Despite the high level of attention on infectious respiratory aerosol during the Covid19 pandemic, little is known about the processes how these particles form inside the respiratory system. Understanding the underlying fluid mechanical processes and their influencing factors would enable the development of drugs to suppress the generation of infectious aerosol. In the proposed work, we focus on the shear-induced mechanism of aerosol generation, which is supposed to occur mostly in the larger airways during coughing. In this process, high air velocities trigger Kelvin-Helmholtz waves in the mucus film, which lines the air vessels. Through a series of instabilities, particles detach from the crest of these waves. In the proposed work, we investigate the process of shear-induced aerosol generation in idealized experiments where we vary the air-flow characteristics and the properties of the mucus fluid. Our central aim here is to deduct an empirical model of the quantity and size distribution of generated particles depending on the mucus rheology and the local shear flow. Further, we observe the wave topology to better understand the coupling between the air flow and the waves. In our experimental setup, we measure the quantity of created particles and the emerging waves simultaneously. To ensure controllable conditions, we simplify the complex flow conditions in the airways. We use a rectangular channel with the bottom wall covered in a mucus mimetic. The mucus mimetic fluid is a synthetic hydrogel developed to recreate the viscoelastic properties and low surface tensions of the mucus. Filtered pressurized air is guided through the channel to trigger shear-induced aerosol generation. After passing the mucus mimetic, the air enters into a collection chamber from where particles are sampled continuously by an aerosol spectrometer. To measure wave topology, we use planar laser induced fluorescence. For this, we stain the mucus mimetic with fluorescent dyes and illuminate a line on the surface of the fluid film with a 532 nm laser. A high-resolution camera captures the resulting fluorescent glow of the mucus mimetic. Figure 1 presents exemplary wave topology results from the experiments, employing varying air flow volume rates, mucus mimetic gel properties, and different configurations of the laser and camera. The resulting wave topologies exhibit significant variation. For the conference, we will conduct parameter studies of the particle quantities and wave topology while varying the mucus mimetic properties and the flow rate of the air. Additionally, we will present grid projection-based techniques to extend the single-line wave measurements and asses the entire surface of the mucus film.}, language = {en} } @article{KranzWagnerPointneretal., author = {Kranz, Michael and Wagner, Maria Stella and Pointner, Daniel and Haus, Moritz and Lubnow, Matthias and Lehle, Karla and Krenkel, Lars}, title = {Polymer embedding of membrane lungs for histological investigations of intra-device clot formation}, series = {Cardiovascular Medicine}, volume = {13}, journal = {Cardiovascular Medicine}, publisher = {Frontiers}, address = {Lausanne}, doi = {10.3389/fcvm.2026.1650978}, pages = {21}, abstract = {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}, language = {en} } @article{BartschEigenbergerFelthausetal., author = {Bartsch, Alexander and Eigenberger, Andreas and Felthaus, Oliver and Booth, Brian W. and Krenkel, Lars and Schratzenstaller, Thomas and Prantl, Lukas}, title = {Quantification of Cannula Influence on Negative Pressure in Syringe Assisted Liposuction}, series = {Plastic \& Reconstructive Surgery}, journal = {Plastic \& Reconstructive Surgery}, publisher = {American Society of Plastic Surgeons}, issn = {0032-1052}, doi = {10.1097/PRS.0000000000012477}, abstract = {To make the results of the autologous fat transfer more satisfactory, all parameters of suction, preparation, and reapplication must be well understood. Since syringe-assisted liposuction is frequently used for small-volume procedures, we quantified the influence of the cannula on negative pressure. For these experiments, we used a digital manometer to which a syringe and one of seven different cannulas were connected. Data was collected and analyzed using LabVIEW and Matlab. The volume of our measurement setup was calculated using Boyle-Mariotte's law to ease interpretation, which revealed a volume of 2.82 ± 0.03 mL. We demonstrate that the volume of the measurement setup significantly affects the measurement results. Consequently, we accounted for this factor and calculated the theoretical values accordingly. Statistical analysis shows a significant influence on negative pressure across different cannulas, except for two pairs, which had minimal volume differences. Exemplary, for cannulas with volumes under 1.75 mL, a 2 mL syringe plunger pull was sufficient to create a - 0.5 bar (- 14.8 inHg) negative pressure. The present data indicate that the size of the suction cannula must be considered in syringe-assisted suction to ensure a specific negative pressure is not exceeded. Other research groups have demonstrated that adipocyte vitality and the amount of stromal vascular fraction are reduced by excessively strong negative pressure.}, language = {en} } @article{DeuterHajBrawanskietal., author = {Deuter, Daniel and Haj, Amer and Brawanski, Alexander and Krenkel, Lars and Schmidt, Nils Ole and Doenitz, Christian}, title = {Fast simulation of hemodynamics in intracranial aneurysms for clinical use}, series = {Acta Neurochirurgica}, volume = {167}, journal = {Acta Neurochirurgica}, publisher = {Springer}, doi = {10.1007/s00701-025-06469-9}, pages = {14}, abstract = {BACKGROUND: A widely accepted tool to assess hemodynamics, one of the most important factors in aneurysm pathophysiology, is Computational Fluid Dynamics (CFD). As current workflows are still time consuming and difficult to operate, CFD is not yet a standard tool in the clinical setting. There it could provide valuable information on aneurysm treatment, especially regarding local risks of rupture, which might help to optimize the individualized strategy of neurosurgical dissection during microsurgical aneurysm clipping. METHOD: We established and validated a semi-automated workflow using 3D rotational angiographies of 24 intracranial aneurysms from patients having received aneurysm treatment at our centre. Reconstruction of vessel geometry and generation of volume meshes was performed using AMIRA 6.2.0 and ICEM 17.1. For solving ANSYS CFX was used. For validational checks, tests regarding the volumetric impact of smoothing operations, the impact of mesh sizes on the results (grid convergence), geometric mesh quality and time tests for the time needed to perform the workflow were conducted in subgroups. RESULTS: Most of the steps of the workflow were performed directly on the 3D images requiring no programming experience. The workflow led to final CFD results in a mean time of 22 min 51.4 s (95\%-CI 20 min 51.562 s-24 min 51.238 s, n = 5). Volume of the geometries after pre-processing was in mean 4.46\% higher than before in the analysed subgroup (95\%-CI 3.43-5.50\%). Regarding mesh sizes, mean relative aberrations of 2.30\% (95\%-CI 1.51-3.09\%) were found for surface meshes and between 1.40\% (95\%-CI 1.07-1.72\%) and 2.61\% (95\%-CI 1.93-3.29\%) for volume meshes. Acceptable geometric mesh quality of volume meshes was found. CONCLUSIONS: We developed a semi-automated workflow for aneurysm CFD to benefit from hemodynamic data in the clinical setting. The ease of handling opens the workflow to clinicians untrained in programming. As previous studies have found that the distribution of hemodynamic parameters correlates with thin-walled aneurysm areas susceptible to rupture, these data might be beneficial for the operating neurosurgeon during aneurysm surgery, even in acute cases.}, language = {en} } @article{PaschkeFoltanWagneretal., author = {Paschke, Lukas and Foltan, Maik and Wagner, Maria S. and Lubnow, Matthias and Gruber, Michael and Krenkel, Lars and Lehle, Karla}, title = {Clinical Relevance of Platelet-Leukocyte Aggregates and Platelet P-Selectin Expression During Venovenous Extracorporeal Membrane Oxygenation}, series = {ASAIO Journal}, journal = {ASAIO Journal}, number = {April 03}, publisher = {Wolters Kluwer}, issn = {1058-2916}, doi = {10.1097/MAT.0000000000002421}, abstract = {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.}, language = {en} } @article{KrieglKrenkelShamoninChamonine, author = {Kriegl, Raphael and Krenkel, Lars and Shamonin (Chamonine), Mikhail}, title = {Preservation of wetting ridges using field-induced plasticity of magnetoactive elastomers}, series = {Journal of Colloid and Interface Science}, volume = {683}, journal = {Journal of Colloid and Interface Science}, publisher = {Elsevier}, issn = {0021-9797}, doi = {10.1016/j.jcis.2024.12.132}, pages = {1019 -- 1027}, abstract = {Hypothesis. The presence of a wetting ridge is crucial for many wetting phenomena on soft substrates. Conventional experimental observations of a wetting ridge require permanent presence of a droplet. The magnetic field-induced plasticity effect (FIPE) of soft magnetoative elastomers (MAEs) allows one to overcome this limitation. Depositing a droplet onto an MAE surface and applying a magnetic field fixes the wetting ridge in place due to the FIPE. The droplet can be removed to investigate the ridge with ease using conventional optical methods. Experiments. The wetting ridge is observed on MAEs with different shear moduli and different material thicknesses by confocal laser scanning microscopy (LSM). The preservation quality of the ridge is analyzed for several values of magnetic field from 10 mT to 270 mT. To verify the plausibilty of results obtained, the measured shape is compared to a theoretical model by Style \& Dufresne. Findings. Upon removal of the sessile droplet, the deformation remains embossed onto the MAE surface as long as the magnetic field remains applied. The height of the ridge peak on soft samples (effective shear modulus kPa) is close to the theoretical prediction for a magnetic flux density of 50 mT. The magnitude of the external magnetic field magnifies the height of the wetting ridge.}, language = {en} } @article{LehlePhilippKrenkeletal., author = {Lehle, Karla and Philipp, Alois and Krenkel, Lars and Gruber, Michael and Hiller, Karl-Anton and M{\"u}ller, Thomas and Lubnow, Matthias}, title = {Thrombocytopenia During Venovenous Extracorporeal Membrane Oxygenation in Adult Patients With Bacterial, Viral, and COVID-19 Pneumonia}, series = {ASAIO Journal}, journal = {ASAIO Journal}, publisher = {Wolters Kluwer}, issn = {1058-2916}, doi = {10.1097/MAT.0000000000002383}, abstract = {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.}, language = {en} } @article{FoltanDinhGruberetal., author = {Foltan, Maik and Dinh, D. and Gruber, Michael and M{\"u}ller, Thomas and Hart, C. and Krenkel, Lars and Schmid, C. and Lehle, Karla}, title = {Incidence of neutrophil extracellular traps (NETs) in different membrane oxygenators: pilot in vitro experiments in commercially available coated membranes}, series = {Journal of artificial organs : the official journal of the Japanese Society for Artificial Organs}, journal = {Journal of artificial organs : the official journal of the Japanese Society for Artificial Organs}, doi = {10.1007/s10047-024-01486-4}, abstract = {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.}, language = {en} } @unpublished{ThausHofrichterLubnowetal., author = {Thaus, Christopher and Hofrichter, Elena and Lubnow, Matthias and Krenkel, Lars and Lehle, Karla}, title = {Hemocompatibility of Membrane Lung Components from Extracorporeal Membrane Oxygenation with Different Antithrombogenic Coatings}, doi = {10.20944/preprints202412.0615.v1}, pages = {1 -- 11}, abstract = {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 na{\"i}ve 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.}, language = {en} } @inproceedings{RuettenKrenkelQuadrio, author = {R{\"u}tten, Markus and Krenkel, Lars and Quadrio, Maurizio}, title = {Simulation and Analyis of the Unsteady Flow within Nasal Airways}, series = {9th European Congress on Computational Methods in Applied Sciences and Engineering - ECCOMAS Congress, 3-7 June 2024, Lisbon, Portugal}, booktitle = {9th European Congress on Computational Methods in Applied Sciences and Engineering - ECCOMAS Congress, 3-7 June 2024, Lisbon, Portugal}, language = {en} } @article{HausFoltanPhilippetal., author = {Haus, Moritz and Foltan, Maik and Philipp, Alois and M{\"u}ller, Thomas and Lingel, Maximilian P. and Krenkel, Lars and Gruber, Michael and Lehle, Karla}, title = {Neutrophil extracellular traps -a potential trigger for the development of thrombocytopenia during extracorporeal membrane oxygenation}, volume = {15}, publisher = {frontiers}, doi = {10.3389/fimmu.2024.1339235}, abstract = {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.}, language = {en} } @article{WagnerKranzKrenkeletal., author = {Wagner, Maria Stella and Kranz, Michael and Krenkel, Lars and Pointner, Daniel and Foltan, Maik and Lubnow, Matthias and Lehle, Karla}, title = {Computer based visualization of clot structures in extracorporeal membrane oxygenation and histological clot investigations for understanding thrombosis in membrane lungs}, series = {Frontiers in Medicine}, journal = {Frontiers in Medicine}, number = {11}, editor = {Becatti, Matteo}, publisher = {Frontiers}, doi = {10.3389/fmed.2024.1416319}, abstract = {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.}, language = {en} }