Mimicking the Liver Sinusoidal Endothelial Cell Niche In Vitro to Enhance Fenestration in a Genetic Model of Systemic Inflammation
Liver sinusoidal endothelial cells (LSECs) play a crucial role in hepatic homeostasis, clearance, and microcirculatory regulation. Their fenestrations—patent transcellular pores—are essential for proper liver function, yet disappear in pathological conditions such as liver fibrosis and inflammation...
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2025-04-01
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| author | Dibakar Borah Oliwia Blacharczyk Karolina Szafranska Izabela Czyzynska-Cichon Sara Metwally Konrad Szymanowski Wolfgang Hübner Jerzy Kotlinowski Ewelina Dobosz Peter McCourt Thomas Huser Malgorzata Lekka Bartlomiej Zapotoczny |
| author_facet | Dibakar Borah Oliwia Blacharczyk Karolina Szafranska Izabela Czyzynska-Cichon Sara Metwally Konrad Szymanowski Wolfgang Hübner Jerzy Kotlinowski Ewelina Dobosz Peter McCourt Thomas Huser Malgorzata Lekka Bartlomiej Zapotoczny |
| author_sort | Dibakar Borah |
| collection | DOAJ |
| description | Liver sinusoidal endothelial cells (LSECs) play a crucial role in hepatic homeostasis, clearance, and microcirculatory regulation. Their fenestrations—patent transcellular pores—are essential for proper liver function, yet disappear in pathological conditions such as liver fibrosis and inflammation through a process known as defenestration. Defenestrated sinusoids are often linked to the liver stiffening that occurs through mechanotransduction-regulated processes. We performed a detailed characterization of polyacrylamide (PAA) hydrogels using atomic force microscopy (AFM), rheometry, scanning electron microscopy, and fluorescence microscopy to assess their potential as biomimetic substrates for LSECs. We additionally implemented AFM; quantitative fluorescence microscopy, including high-resolution structured illumination microscopy (HR-SIM); and an endocytosis assay to characterize the morphology and function of LSECs. Our results revealed significant local variations in hydrogel stiffness and differences in pore sizes. The primary LSECs cultured on these substrates had a range of stiffnesses and were analyzed with regard to their number of fenestrations, cytoskeletal organization, and endocytic function. To explore mechanotransduction in inflammatory liver disease, we investigated LSECs from a genetic model of systemic inflammation triggered by the deletion of Mcpip1 in myeloid leukocytes and examined their ability to restore their fenestrations on soft substrates. Our study demonstrates the beneficial effect of soft hydrogels on LSECs. Control cells exhibited a similar fenestrated morphology and function compared to cells cultured on plastic substrates. However, the pathological LSECs from the genetic model of systemic inflammation regained their fenestrations when cultured on soft hydrogels. This observation supports previous findings on the beneficial effects of soft substrates on LSEC fenestration status. |
| format | Article |
| id | doaj-art-540b9fc1f3fc4493b6474b95fa2adc68 |
| institution | DOAJ |
| issn | 2073-4409 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | MDPI AG |
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| series | Cells |
| spelling | doaj-art-540b9fc1f3fc4493b6474b95fa2adc682025-08-20T03:14:17ZengMDPI AGCells2073-44092025-04-0114862110.3390/cells14080621Mimicking the Liver Sinusoidal Endothelial Cell Niche In Vitro to Enhance Fenestration in a Genetic Model of Systemic InflammationDibakar Borah0Oliwia Blacharczyk1Karolina Szafranska2Izabela Czyzynska-Cichon3Sara Metwally4Konrad Szymanowski5Wolfgang Hübner6Jerzy Kotlinowski7Ewelina Dobosz8Peter McCourt9Thomas Huser10Malgorzata Lekka11Bartlomiej Zapotoczny12Department of Biophysical Microstructures, Institute of Nuclear Physics, Polish Academy of Sciences, ul. Radzikowskiego 142, 31-342 Krakow, PolandDepartment of Biophysical Microstructures, Institute of Nuclear Physics, Polish Academy of Sciences, ul. Radzikowskiego 142, 31-342 Krakow, PolandVascular Biology Research Group, Department of Medical Biology University of Tromsø—The Arctic University of Norway, 9019 Tromsø, NorwayJagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, Bobrzynskiego 14, 30-348 Krakow, PolandDepartment of Biophysical Microstructures, Institute of Nuclear Physics, Polish Academy of Sciences, ul. Radzikowskiego 142, 31-342 Krakow, PolandDepartment of Biophysical Microstructures, Institute of Nuclear Physics, Polish Academy of Sciences, ul. Radzikowskiego 142, 31-342 Krakow, PolandBiomolecular Photonics, Faculty of Physics, Bielefeld University, 33615 Bielefeld, GermanyDepartment of General Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Krakow, PolandDepartment of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Krakow, PolandVascular Biology Research Group, Department of Medical Biology University of Tromsø—The Arctic University of Norway, 9019 Tromsø, NorwayBiomolecular Photonics, Faculty of Physics, Bielefeld University, 33615 Bielefeld, GermanyDepartment of Biophysical Microstructures, Institute of Nuclear Physics, Polish Academy of Sciences, ul. Radzikowskiego 142, 31-342 Krakow, PolandDepartment of Biophysical Microstructures, Institute of Nuclear Physics, Polish Academy of Sciences, ul. Radzikowskiego 142, 31-342 Krakow, PolandLiver sinusoidal endothelial cells (LSECs) play a crucial role in hepatic homeostasis, clearance, and microcirculatory regulation. Their fenestrations—patent transcellular pores—are essential for proper liver function, yet disappear in pathological conditions such as liver fibrosis and inflammation through a process known as defenestration. Defenestrated sinusoids are often linked to the liver stiffening that occurs through mechanotransduction-regulated processes. We performed a detailed characterization of polyacrylamide (PAA) hydrogels using atomic force microscopy (AFM), rheometry, scanning electron microscopy, and fluorescence microscopy to assess their potential as biomimetic substrates for LSECs. We additionally implemented AFM; quantitative fluorescence microscopy, including high-resolution structured illumination microscopy (HR-SIM); and an endocytosis assay to characterize the morphology and function of LSECs. Our results revealed significant local variations in hydrogel stiffness and differences in pore sizes. The primary LSECs cultured on these substrates had a range of stiffnesses and were analyzed with regard to their number of fenestrations, cytoskeletal organization, and endocytic function. To explore mechanotransduction in inflammatory liver disease, we investigated LSECs from a genetic model of systemic inflammation triggered by the deletion of Mcpip1 in myeloid leukocytes and examined their ability to restore their fenestrations on soft substrates. Our study demonstrates the beneficial effect of soft hydrogels on LSECs. Control cells exhibited a similar fenestrated morphology and function compared to cells cultured on plastic substrates. However, the pathological LSECs from the genetic model of systemic inflammation regained their fenestrations when cultured on soft hydrogels. This observation supports previous findings on the beneficial effects of soft substrates on LSEC fenestration status.https://www.mdpi.com/2073-4409/14/8/621liver sinusoidal endothelial cellspolyacrylamideatomic force microscopyelastic propertiesfenestrationsactin cytoskeleton |
| spellingShingle | Dibakar Borah Oliwia Blacharczyk Karolina Szafranska Izabela Czyzynska-Cichon Sara Metwally Konrad Szymanowski Wolfgang Hübner Jerzy Kotlinowski Ewelina Dobosz Peter McCourt Thomas Huser Malgorzata Lekka Bartlomiej Zapotoczny Mimicking the Liver Sinusoidal Endothelial Cell Niche In Vitro to Enhance Fenestration in a Genetic Model of Systemic Inflammation Cells liver sinusoidal endothelial cells polyacrylamide atomic force microscopy elastic properties fenestrations actin cytoskeleton |
| title | Mimicking the Liver Sinusoidal Endothelial Cell Niche In Vitro to Enhance Fenestration in a Genetic Model of Systemic Inflammation |
| title_full | Mimicking the Liver Sinusoidal Endothelial Cell Niche In Vitro to Enhance Fenestration in a Genetic Model of Systemic Inflammation |
| title_fullStr | Mimicking the Liver Sinusoidal Endothelial Cell Niche In Vitro to Enhance Fenestration in a Genetic Model of Systemic Inflammation |
| title_full_unstemmed | Mimicking the Liver Sinusoidal Endothelial Cell Niche In Vitro to Enhance Fenestration in a Genetic Model of Systemic Inflammation |
| title_short | Mimicking the Liver Sinusoidal Endothelial Cell Niche In Vitro to Enhance Fenestration in a Genetic Model of Systemic Inflammation |
| title_sort | mimicking the liver sinusoidal endothelial cell niche in vitro to enhance fenestration in a genetic model of systemic inflammation |
| topic | liver sinusoidal endothelial cells polyacrylamide atomic force microscopy elastic properties fenestrations actin cytoskeleton |
| url | https://www.mdpi.com/2073-4409/14/8/621 |
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