scholarly journals HepaChip-MP – a twenty-four chamber microplate for a continuously perfused liver coculture model

Lab on a Chip ◽  
2020 ◽  
Vol 20 (16) ◽  
pp. 2911-2926
Author(s):  
Marius Busche ◽  
Olena Tomilova ◽  
Julia Schütte ◽  
Simon Werner ◽  
Meike Beer ◽  
...  
Keyword(s):  
Perfused Liver ◽  
Culture Chamber ◽  
Liver Sinusoid ◽  
Coculture Model

HepaChip-MP: a 24-culture-chamber, automated microfluidic in vitro model of the liver sinusoid in multiwellplate format.

scholarly journals New In Vitro Coculture Model for Evaluating Intestinal Absorption of Different Lipid Nanocapsules

Pharmaceutics ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 595
Author(s):  
Norraseth Kaeokhamloed ◽  
Emillie Roger ◽  
Jérôme Béjaud ◽  
Nolwenn Lautram ◽  
Florence Manero ◽  
...  
Keyword(s):  
Surface Modification ◽  
Intestinal Absorption ◽  
Oral Absorption ◽  
Membrane Integrity ◽  
Microvascular Endothelial Cell ◽  
Absorption Mechanism ◽  
Apparent Permeability ◽  
Lipid Nanocapsules ◽  
Coculture Model

Standard models used for evaluating the absorption of nanoparticles like Caco-2 ignore the presence of vascular endothelium, which is a part of the intestinal multi-layered barrier structure. Therefore, a coculture between the Caco-2 epithelium and HMEC-1 (Human Microvascular Endothelial Cell type 1) on a Transwell® insert has been developed. The model has been validated for (a) membrane morphology by transmission electron microscope (TEM); (b) ZO-1 and β-catenin expression by immunoassay; (c) membrane integrity by trans-epithelial electrical resistance (TEER) measurement; and (d) apparent permeability of drugs from different biopharmaceutical classification system (BCS) classes. Lipid nanocapsules (LNCs) were formulated with different sizes (55 and 85 nm) and surface modifications (DSPE-mPEG (2000) and stearylamine). Nanocapsule integrity and particle concentration were monitored using the Förster resonance energy transfer (FRET) technique. The result showed that surface modification by DSPE-mPEG (2000) increased the absorption of 55-nm LNCs in the coculture model but not in the Caco-2. Summarily, the coculture model was validated as a tool for evaluating the intestinal absorption of drugs and nanoparticles. The new coculture model has a different LNCs absorption mechanism suggesting the importance of intestinal endothelium and reveals that the surface modification of LNCs can modify the in vitro oral absorption.

Effects of peritoneal macrophages from women with endometriosis on endometrial cellular proliferation in an in vitro coculture model

1999 ◽  
Vol 72 (3) ◽  
pp. 533-538 ◽  
Author(s):  
Foo-Hoe Loh ◽  
Ariff Bongso ◽  
Chui-Yee Fong ◽  
Dow-Rhoon Koh ◽  
Szu-Hee Lee ◽  
...  
Keyword(s):  
Cellular Proliferation ◽  
Peritoneal Macrophages ◽  
Coculture Model

scholarly journals Monocyte-induced recovery of inflammation-associated hepatocellular dysfunction in a biochip-based human liver model

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Marko Gröger ◽  
Knut Rennert ◽  
Benjamin Giszas ◽  
Elisabeth Weiß ◽  
Julia Dinger ◽  
...  
Keyword(s):  
Liver Dysfunction ◽  
Human Liver ◽  
Immune Cell ◽  
Early Event ◽  
Liver Sinusoid ◽  
Human Sepsis ◽  
Cellular Events ◽  
Cell Layers ◽  
Parenchymal Cells

Abstract Liver dysfunction is an early event in sepsis-related multi-organ failure. We here report the establishment and characterization of a microfluidically supported in vitro organoid model of the human liver sinusoid. The liver organoid is composed of vascular and hepatocyte cell layers integrating non-parenchymal cells closely reflecting tissue architecture and enables physiological cross-communication in a bio-inspired fashion. Inflammation-associated liver dysfunction was mimicked by stimulation with various agonists of toll-like receptors. TLR-stimulation induced the release of pro- and anti-inflammatory cytokines and diminished expression of endothelial VE-cadherin, hepatic MRP-2 transporter and apolipoprotein B (ApoB), resulting in an inflammation-related endothelial barrier disruption and hepatocellular dysfunction in the liver organoid. However, interaction of the liver organoid with human monocytes attenuated inflammation-related cell responses and restored MRP-2 transporter activity, ApoB expression and albumin/urea production. The cellular events observed in the liver organoid closely resembled pathophysiological responses in the well-established sepsis model of peritoneal contamination and infection (PCI) in mice and clinical observations in human sepsis. We therefore conclude that this human liver organoid model is a valuable tool to investigate sepsis-related liver dysfunction and subsequent immune cell-related tissue repair/remodeling processes.

scholarly journals Role of nitric oxide in heparin-induced attenuation of hypoxic pulmonary vascular remodeling

2002 ◽  
Vol 92 (5) ◽  
pp. 2012-2018 ◽  
Author(s):  
Damian J. Horstman ◽  
Lars G. Fischer ◽  
Peter C. Kouretas ◽  
Robert L. Hannan ◽  
George F. Rich
Keyword(s):  
Nitric Oxide ◽  
Vascular Remodeling ◽  
Pulmonary Vasculature ◽  
Pulmonary Vascular Remodeling ◽  
Antiproliferative Effects ◽  
Coculture Model ◽  
Pulmonary Arterial ◽  
Nos Inhibitor

Heparin and nitric oxide (NO) attenuate changes to the pulmonary vasculature caused by prolonged hypoxia. Heparin may increase NO; therefore, we hypothesized that heparin may attenuate hypoxia-induced pulmonary vascular remodeling via a NO-mediated mechanism. In vivo, rats were exposed to normoxia (N) or hypoxia (H; 10% O2) with or without heparin (1,200 U · kg−1 · day−1) and/or the NO synthase (NOS) inhibitor N ω-nitro-l-arginine methyl ester (l-NAME; 20 mg · kg−1 · day−1) for 3 days or 3 wk. Heparin attenuated increases in pulmonary arterial pressure, the percentage of muscular pulmonary vessels, and their medial thickness induced by 3 wk of H. Importantly, althoughl-NAME alone had no effect, it prevented these effects of heparin on vascular remodeling. In H lungs, heparin increased NOS activity and cGMP levels at 3 days and 3 wk and endothelial NOS protein expression at 3 days but not at 3 wk. In vitro, heparin (10 and 100 U · kg−1 · ml−1) increased cGMP levels after 10 min and 24 h in N and anoxic (0% O2) endothelial cell-smooth muscle cell (SMC) coculture. SMC proliferation, assessed by 5-bromo-2′-deoxyuridine incorporation during a 3-h incubation period, was decreased by heparin under N, but not anoxic, conditions. The antiproliferative effects of heparin were not altered byl-NAME. In conclusion, the in vivo results suggest that attenuation of hypoxia-induced pulmonary vascular remodeling by heparin is NO mediated. Heparin increases cGMP in vitro; however, the heparin-induced decrease in SMC proliferation in the coculture model appears to be NO independent.

scholarly journals Anti-Inflammatory and Antiosteoclastogenic Activities of Parthenolide on Human Periodontal Ligament CellsIn Vitro

2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
Xufang Zhang ◽  
Chen Fan ◽  
Yin Xiao ◽  
Xueli Mao
Keyword(s):  
Periodontal Ligament ◽  
Tooth Loss ◽  
Nuclear Factor Kappa B ◽  
Raw264.7 Cells ◽  
Erk Activation ◽  
Tanacetum Parthenium ◽  
Coculture Model ◽  
Anti Inflammatory ◽  
Potential Use

Periodontitis is an inflammatory disease that causes osteolysis and tooth loss. It is known that the nuclear factor kappa B (NF-κB) signalling pathway plays a key role in the progression of inflammation and osteoclastogenesis in periodontitis. Parthenolide (PTL), a sesquiterpene lactone extracted from the shoots ofTanacetum parthenium, has been shown to possess anti-inflammatory properties in various diseases. In the study reported herein, we investigated the effects of PTL on the inflammatory and osteoclastogenic response of human periodontal ligament-derived cells (hPDLCs) and revealed the signalling pathways in this process. Our results showed that PTL decreased NF-κB activation, I-κB degradation, and ERK activation in hPDLCs. PTL significantly reduced the expression of inflammatory (IL-1β, IL-6, and TNF-α) and osteoclastogenic (RANKL, OPG, and M-CSF) genes in LPS-stimulated hPDLCs. In addition, PTL attenuated hPDLC-induced osteoclastogenic differentiation of macrophages (RAW264.7 cells), as well as reducing gene expression of osteoclast-related markers in RAW264.7 cells in an hPDLC-macrophage coculture model. Taken together, these results demonstrate the anti-inflammatory and antiosteoclastogenic activities of PTL in hPDLCsin vitro. These data offer fundamental evidence supporting the potential use of PTL in periodontitis treatment.

Targeting IL-17 attenuates hypoxia-induced pulmonary hypertension through downregulation of β-catenin

Thorax ◽  
2019 ◽  
Vol 74 (6) ◽  
pp. 564-578 ◽  
Author(s):  
Lei Wang ◽  
Jie Liu ◽  
Wang Wang ◽  
Xianmei Qi ◽  
Ying Wang ◽  
...  
Keyword(s):  
Pulmonary Hypertension ◽  
Lung Tissue ◽  
Interleukin 17 ◽  
Antibody Assays ◽  
Coculture Model ◽  
Potential Benefits ◽  
Pulmonary Arterial ◽  
And Migration ◽  
Arterial Endothelial Cells

BackgroundThe role of interleukin 17 (IL-17) in hypoxic pulmonary hypertension (HPH) remains unclear. This study is designed to explore whether IL-17 is a potential target for HPH treatment.MethodsClinic samples from the lung tissue and serum were obtained from qualified patients. Western blotting, immunohistochemistry and/or ELISA were used to measure the expression of relevant proteins. HPH models were established in C57BL/6 wild-type (WT) and IL-17−/− mice and were treated with exogenous recombinant mouse IL-17 (rmIL-17) or an IL-17 neutralising antibody. Assays for cell proliferation, angiogenesis and adhesion were employed to analyse the behaviours of human pulmonary arterial endothelial cells (HPAECs). A non-contact Transwell coculture model was used to evaluate intercellular interactions.ResultsExpression of IL-17 was increased in lung tissue of both patients with bronchiectasis/COPD-associated PH and HPH mouse model. Compared with WT mice, IL-17−/− mice had attenuated HPH, whereas administration of rmIL-17 aggravated HPH. In vitro, recombinant human IL-17 (rhIL-17) promoted proliferation, angiogenesis and adhesion in HPAECs through upregulation of Wnt3a/β-catenin/CyclinD1 pathway, and siRNA-mediated knockdown of β-catenin almost completely reversed this IL-17-mediated phenomena. IL-17 promoted the proliferation but not the migration of human pulmonary arterial smooth muscle cells (HPASMCs) cocultured with HPAECs under both normoxia and hypoxia, but IL-17 had no direct effect on proliferation and migration of HPASMCs. Blockade of IL-17 with a neutralising antibody attenuated HPH in WT mice.ConclusionsIL-17 contributes to the pathogenesis of HPH through upregulation of β-catenin expression. Targeting IL-17 might provide potential benefits for alternative therapeutic strategies for HPH.

Stimulus of leucine incorporation into perfused liver protein by insulin

1963 ◽  
Vol 204 (1) ◽  
pp. 140-142 ◽  
Author(s):  
J. C. Penhos ◽  
M. E. Krahl
Keyword(s):  
Blood Sugar ◽  
Total Concentration ◽  
Diabetic Rats ◽  
Leucine Incorporation ◽  
Liver Protein ◽  
Perfused Liver ◽  
Partial Pancreatectomy ◽  
Perfusion Period ◽  
Terminal Blood

Livers from 24-hr fasting sham-operated or partially pancreatectomized rats were perfused at 38 C with heparinized blood from normal 24-hr fasting rats. Leucine-1-C14, 1.6 x 105 counts/min per ml, and glucose to make the initial total concentration 2 mg/ml were added to the blood. Incorporation of leucine-1-C14 into protein proceeded linearly for 2 hr, then at a decreased rate. The leucine-1-C14 incorporated into protein over the initial 2-hr perfusion period decreased progressively as the time after partial pancreatectomy of the liver donors increased, up to 6 months. Insulin, 0.01 unit/ml, added to the perfusing blood stimulated leucine incorporation into protein of livers from rats partially pancreatectomized 3 months before (mean terminal blood sugar 160 mg/100 ml) but not into that of livers from the 6-month series (mean terminal blood sugar 315 mg/100 ml). No significant effect of insulin was obtained in any of the sham-operated groups. It is concluded that insulin added in vitro can stimulate incorporation of leucine into protein in livers from mildly diabetic, but not severely diabetic, rats.

scholarly journals IFN-γ enhances cell-mediated cytotoxicity against keratinocytes via JAK2/STAT1 in lichen planus

2019 ◽  
Vol 11 (511) ◽  
pp. eaav7561 ◽  
Author(s):  
Shuai Shao ◽  
Lam C. Tsoi ◽  
Mrinal K. Sarkar ◽  
Xianying Xing ◽  
Ke Xue ◽  
...  
Keyword(s):  
Lichen Planus ◽  
Therapeutic Agents ◽  
Janus Kinase ◽  
Unknown Etiology ◽  
Type Ii ◽  
Jak Inhibitors ◽  
Coculture Model ◽  
Ifn Γ ◽  
Cytotoxic Responses

Lichen planus (LP) is a chronic debilitating inflammatory disease of unknown etiology affecting the skin, nails, and mucosa with no current FDA-approved treatments. It is histologically characterized by dense infiltration of T cells and epidermal keratinocyte apoptosis. Using global transcriptomic profiling of patient skin samples, we demonstrate that LP is characterized by a type II interferon (IFN) inflammatory response. The type II IFN, IFN-γ, is demonstrated to prime keratinocytes and increase their susceptibility to CD8+ T cell–mediated cytotoxic responses through MHC class I induction in a coculture model. We show that this process is dependent on Janus kinase 2 (JAK2) and signal transducer and activator of transcription 1 (STAT1), but not JAK1 or STAT2 signaling. Last, using drug prediction algorithms, we identify JAK inhibitors as promising therapeutic agents in LP and demonstrate that the JAK1/2 inhibitor baricitinib fully protects keratinocytes against cell-mediated cytotoxic responses in vitro. In summary, this work elucidates the role and mechanisms of IFN-γ in LP pathogenesis and provides evidence for the therapeutic use of JAK inhibitors to limit cell-mediated cytotoxicity in patients with LP.

VSM growth is stimulated in sympathetic neuron/VSM cocultures: role of TGF-β2 and endothelin

2000 ◽  
Vol 278 (2) ◽  
pp. H404-H411 ◽  
Author(s):  
Deborah H. Damon
Keyword(s):  
Transforming Growth Factor ◽  
Sympathetic Neurons ◽  
Sympathetic Nerves ◽  
Sympathetic Neuron ◽  
Vessel Growth ◽  
Coculture Model ◽  
Transforming Growth Factor Β2 ◽  
Cervical Ganglia ◽  
Stimulation Of

Sympathetic nerves are purported to stimulate blood vessel growth. The mechanism(s) underlying this stimulation has not been determined. With use of an in vitro coculture model, the present study tests the hypothesis that sympathetic neurons stimulate the growth of vascular smooth muscle (VSM) and evaluates potential mechanisms mediating this stimulation. Sympathetic neurons isolated from superior cervical ganglia (SCG) stimulated the growth of VSM. Growth of VSM in the presence of SCG (856 ± 81%) was significantly greater than that in the absence of SCG (626 ± 66%, P < 0.05). SCG did not stimulate VSM growth in transwell cocultures. An antibody that neutralized the activity of transforming growth factor-β2 (TGF-β2) inhibited SCG stimulation of VSM growth in coculture. SCG stimulation of VSM growth was also inhibited by an endothelin A receptor antagonist. These data suggest novel mechanisms for sympathetic modulation of vascular growth that may play a role in the physiological and/or pathological growth of the vasculature.

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