The Experts below are selected from a list of 186 Experts worldwide ranked by ideXlab platform
Geraldine A Hamilton - One of the best experts on this subject based on the ideXlab platform.
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primary human Lung Alveolus on a chip model of intravascular thrombosis for assessment of therapeutics
Clinical Pharmacology & Therapeutics, 2018Co-Authors: Abhishek Jain, Riccardo Barrile, A D Van Der Meer, Akiko Mammoto, Tadanori Mammoto, K De Ceunynck, Omozuanvbo Aisiku, Monicah A Otieno, Calvert Louden, Geraldine A HamiltonAbstract:Pulmonary thrombosis is a significant cause of patient mortality, however, there are no effective in vitro models of thrombi formation in human Lung microvessels, that could also assess therapeutics and toxicology of antithrombotic drugs. Here we show that a microfluidic Lung Alveolus-on-a-chip lined by human primary alveolar epithelium interfaced with endothelium, and cultured under flowing whole blood can be used to perform quantitative analysis of organ-level contributions to inflammation-induced thrombosis. This microfluidic chip recapitulates in vivo responses, including platelet-endothelial dynamics and revealed that lipopolysaccharide (LPS) endotoxin indirectly stimulates intravascular thrombosis by activating the alveolar epithelium, rather than acting directly on endothelium. This model is also used to analyze inhibition of endothelial activation and thrombosis due to a protease activated receptor-1 (PAR-1) antagonist, demonstrating its ability to dissect complex responses and identify antithrombotic therapeutics. Thus, this methodology offers a new approach to study human pathophysiology of pulmonary thrombosis and advance drug development. This article is protected by copyright. All rights reserved.
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Abstract 342: Analysis of Mechanism of a Novel Drug Candidate using an Organ-level Functional Microdevice that Reconstitutes Human Pulmonary Thrombosis
Arteriosclerosis Thrombosis and Vascular Biology, 2016Co-Authors: Abhishek Jain, Riccardo Barrile, A D Van Der Meer, K De Ceunynck, Omozuanvbo Aisiku, Monicah A Otieno, Calvert Louden, Geraldine A Hamilton, Robert Flaumenhaft, Donald E. IngberAbstract:Pulmonary microvascular thrombosis is a catastrophic medical condition and yet, it is very difficult to predict response and study mechanism of action of potential drug candidates to humans. This is partly so because currently available in vitro assays do not recapitulate physiologically-relevant forces and animal models can also be very complex, making it impossible to analyze intercellular signaling within the Lung that occurs under coagulation or drug administration. We designed a model of Lung thrombosis in which human primary alveolar and endothelial cells are co-cultured and maintained up to 2 weeks. The device consists of a top chamber seeded with human alveolar epithelial cells (AE) and a lower chamber seeded with endothelial cells, separated by a porous matrix-coated membrane. Whole blood was perfused at a physiological shear stress through the vascular channel and clots were visualized in real-time. When healthy cells were cultured, no intravascular blood clotting was observed, even when lipopolysaccharide (LPS) endotoxin was administered. In contrast, when LPS was added to the AE channel, it caused a significant increase in platelet adhesion at the endothelium, demonstrating that the presence of alveolar epithelium is critical to LPS-induced intravascular thrombosis in vitro . We evaluated this device by analyzing a novel protease activator receptor-1 (PAR1) antithrombotic compound, termed parmodulin 2 (PM2). When the endothelium was cultured along with PM2 under the condition of LPS stimulated AE, we found inhibition of clotting, demonstrating the therapeutic effect of PM2 in the presence of epithelial-endothelial-blood cell signaling. Finally, to test if PM2 performs the therapeutic function of Activated Protein C (APC) that has been reported to stimulate its cytosolic effects via the β-arrestin pathway, we knocked down β-arrestin in the endothelium and analyzed clot formation again. We found that clotting reoccurred in vascular channel, thus showing that PM2 acts like an APC-like drug candidate. In conclusion, the Lung Alveolus-on-a-chip reconstitutes organ-level responses to blood clotting and may offer a valuable platform for drug development by allowing to dissect contributions of various cells in their mechanism of action.
Changyong Wang - One of the best experts on this subject based on the ideXlab platform.
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the reconstruction of Lung Alveolus like structure in collagen matrigel microcapsules scaffolds in vitro
Journal of Cellular and Molecular Medicine, 2011Co-Authors: Wenjun Zhang, Ye Zhang, Haibin Wang, Yanmeng Wang, Cuimi Duan, Jin Zhou, Changyong WangAbstract:This study attempted to use collagen–Matrigel as extracellular matrix (ECM) to supply cells with three-dimensional (3D) culture condition and employ alginate-poly-l-lysine-alginate (APA) microcapsules to control the formation of Alveolus-like structure in vitro. We tested mice foetal pulmonary cells (FPCs) by immunohistochemistry after 2D culture. The Alveolus-like structure was reconstructed by seeding FPCs in collagen–Matrigel mixed with APA microcapsules 1.5 ml. A self-made mould was used to keep the structure from contraction. Meanwhile, it provided static stretch to the structure. After 7, 14 and 21 days of culture, the Alveolus-like structure was analysed histologically and immunohistochemically, or by scanning transmission electron microscopy (TEM). We also observed these structures under inverted phase contrast microscope. The expression of pro-surfactant protein C (SpC) was detected by reverse transcription-polymerase chain reaction (RT-PCR). We obtained fibroblasts, epithelial cells and alveolar type II (AE2) cells in FPCs. In the reconstructed structure, seeding cells surrounding the APA microcapsules constructed Alveolus-like structures, the size of them ranges from 200 to 300 μm. In each reconstructed Lung tissue sheet, microcapsules had integrity. Pan-cytokeratin, vimentin and SpC positive cells were observed in 7- and 14-day cultured structures. TEM showed lamellar bodies of AE2 cells in the reconstructed tissues whereas RT-PCR expressed SpC gene. Primary mice FPCs could form Alveolus-like structures in collagen–Matrigel/APA microcapsules engineered scaffolds, which could maintain a differentiated state of AE2 cells.
Heber C. Nielsen - One of the best experts on this subject based on the ideXlab platform.
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Neuregulin-ErbB4 signaling in the developing Lung Alveolus: a brief review
Journal of Cell Communication and Signaling, 2014Co-Authors: Najla Fiaturi, John J. Castellot, Heber C. NielsenAbstract:Lung immaturity is the major cause of morbidity and mortality in premature infants, especially those born
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neuregulin erbb4 signaling in the developing Lung Alveolus a brief review
Journal of Cell Communication and Signaling, 2014Co-Authors: Najla Fiaturi, John J. Castellot, Heber C. NielsenAbstract:Lung immaturity is the major cause of morbidity and mortality in premature infants, especially those born <28 weeks gestation. Proper Lung development from 23–28 weeks requires coordinated cell proliferation and differentiation. Infants born at this age are at high risk for respiratory distress syndrome (RDS), a Lung disease characterized by insufficient surfactant production due to immaturity of the alveoli and its constituent cells in the Lung. The ErbB4 receptor and its stimulation by neuregulin (NRG) plays a critical role in surfactant synthesis by alveolar type II epithelial cells. In this review, we first provide an introduction to normal human alveolar development, followed by a discussion of the neuregulin and ErbB4-mediated mechanisms regulating alveolar development and surfactant production.
Wenjun Zhang - One of the best experts on this subject based on the ideXlab platform.
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the reconstruction of Lung Alveolus like structure in collagen matrigel microcapsules scaffolds in vitro
Journal of Cellular and Molecular Medicine, 2011Co-Authors: Wenjun Zhang, Ye Zhang, Haibin Wang, Yanmeng Wang, Cuimi Duan, Jin Zhou, Changyong WangAbstract:This study attempted to use collagen–Matrigel as extracellular matrix (ECM) to supply cells with three-dimensional (3D) culture condition and employ alginate-poly-l-lysine-alginate (APA) microcapsules to control the formation of Alveolus-like structure in vitro. We tested mice foetal pulmonary cells (FPCs) by immunohistochemistry after 2D culture. The Alveolus-like structure was reconstructed by seeding FPCs in collagen–Matrigel mixed with APA microcapsules 1.5 ml. A self-made mould was used to keep the structure from contraction. Meanwhile, it provided static stretch to the structure. After 7, 14 and 21 days of culture, the Alveolus-like structure was analysed histologically and immunohistochemically, or by scanning transmission electron microscopy (TEM). We also observed these structures under inverted phase contrast microscope. The expression of pro-surfactant protein C (SpC) was detected by reverse transcription-polymerase chain reaction (RT-PCR). We obtained fibroblasts, epithelial cells and alveolar type II (AE2) cells in FPCs. In the reconstructed structure, seeding cells surrounding the APA microcapsules constructed Alveolus-like structures, the size of them ranges from 200 to 300 μm. In each reconstructed Lung tissue sheet, microcapsules had integrity. Pan-cytokeratin, vimentin and SpC positive cells were observed in 7- and 14-day cultured structures. TEM showed lamellar bodies of AE2 cells in the reconstructed tissues whereas RT-PCR expressed SpC gene. Primary mice FPCs could form Alveolus-like structures in collagen–Matrigel/APA microcapsules engineered scaffolds, which could maintain a differentiated state of AE2 cells.
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The reconstruction of Lung Alveolus‐like structure in collagen‐matrigel/microcapsules scaffolds in vitro
Journal of Cellular and Molecular Medicine, 2011Co-Authors: Wenjun Zhang, Ye Zhang, Haibin Wang, Yanmeng Wang, Cuimi Duan, Jin ZhouAbstract:This study attempted to use collagen–Matrigel as extracellular matrix (ECM) to supply cells with three-dimensional (3D) culture condition and employ alginate-poly-l-lysine-alginate (APA) microcapsules to control the formation of Alveolus-like structure in vitro. We tested mice foetal pulmonary cells (FPCs) by immunohistochemistry after 2D culture. The Alveolus-like structure was reconstructed by seeding FPCs in collagen–Matrigel mixed with APA microcapsules 1.5 ml. A self-made mould was used to keep the structure from contraction. Meanwhile, it provided static stretch to the structure. After 7, 14 and 21 days of culture, the Alveolus-like structure was analysed histologically and immunohistochemically, or by scanning transmission electron microscopy (TEM). We also observed these structures under inverted phase contrast microscope. The expression of pro-surfactant protein C (SpC) was detected by reverse transcription-polymerase chain reaction (RT-PCR). We obtained fibroblasts, epithelial cells and alveolar type II (AE2) cells in FPCs. In the reconstructed structure, seeding cells surrounding the APA microcapsules constructed Alveolus-like structures, the size of them ranges from 200 to 300 μm. In each reconstructed Lung tissue sheet, microcapsules had integrity. Pan-cytokeratin, vimentin and SpC positive cells were observed in 7- and 14-day cultured structures. TEM showed lamellar bodies of AE2 cells in the reconstructed tissues whereas RT-PCR expressed SpC gene. Primary mice FPCs could form Alveolus-like structures in collagen–Matrigel/APA microcapsules engineered scaffolds, which could maintain a differentiated state of AE2 cells.
Abhishek Jain - One of the best experts on this subject based on the ideXlab platform.
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primary human Lung Alveolus on a chip model of intravascular thrombosis for assessment of therapeutics
Clinical Pharmacology & Therapeutics, 2018Co-Authors: Abhishek Jain, Riccardo Barrile, A D Van Der Meer, Akiko Mammoto, Tadanori Mammoto, K De Ceunynck, Omozuanvbo Aisiku, Monicah A Otieno, Calvert Louden, Geraldine A HamiltonAbstract:Pulmonary thrombosis is a significant cause of patient mortality, however, there are no effective in vitro models of thrombi formation in human Lung microvessels, that could also assess therapeutics and toxicology of antithrombotic drugs. Here we show that a microfluidic Lung Alveolus-on-a-chip lined by human primary alveolar epithelium interfaced with endothelium, and cultured under flowing whole blood can be used to perform quantitative analysis of organ-level contributions to inflammation-induced thrombosis. This microfluidic chip recapitulates in vivo responses, including platelet-endothelial dynamics and revealed that lipopolysaccharide (LPS) endotoxin indirectly stimulates intravascular thrombosis by activating the alveolar epithelium, rather than acting directly on endothelium. This model is also used to analyze inhibition of endothelial activation and thrombosis due to a protease activated receptor-1 (PAR-1) antagonist, demonstrating its ability to dissect complex responses and identify antithrombotic therapeutics. Thus, this methodology offers a new approach to study human pathophysiology of pulmonary thrombosis and advance drug development. This article is protected by copyright. All rights reserved.
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Abstract 342: Analysis of Mechanism of a Novel Drug Candidate using an Organ-level Functional Microdevice that Reconstitutes Human Pulmonary Thrombosis
Arteriosclerosis Thrombosis and Vascular Biology, 2016Co-Authors: Abhishek Jain, Riccardo Barrile, A D Van Der Meer, K De Ceunynck, Omozuanvbo Aisiku, Monicah A Otieno, Calvert Louden, Geraldine A Hamilton, Robert Flaumenhaft, Donald E. IngberAbstract:Pulmonary microvascular thrombosis is a catastrophic medical condition and yet, it is very difficult to predict response and study mechanism of action of potential drug candidates to humans. This is partly so because currently available in vitro assays do not recapitulate physiologically-relevant forces and animal models can also be very complex, making it impossible to analyze intercellular signaling within the Lung that occurs under coagulation or drug administration. We designed a model of Lung thrombosis in which human primary alveolar and endothelial cells are co-cultured and maintained up to 2 weeks. The device consists of a top chamber seeded with human alveolar epithelial cells (AE) and a lower chamber seeded with endothelial cells, separated by a porous matrix-coated membrane. Whole blood was perfused at a physiological shear stress through the vascular channel and clots were visualized in real-time. When healthy cells were cultured, no intravascular blood clotting was observed, even when lipopolysaccharide (LPS) endotoxin was administered. In contrast, when LPS was added to the AE channel, it caused a significant increase in platelet adhesion at the endothelium, demonstrating that the presence of alveolar epithelium is critical to LPS-induced intravascular thrombosis in vitro . We evaluated this device by analyzing a novel protease activator receptor-1 (PAR1) antithrombotic compound, termed parmodulin 2 (PM2). When the endothelium was cultured along with PM2 under the condition of LPS stimulated AE, we found inhibition of clotting, demonstrating the therapeutic effect of PM2 in the presence of epithelial-endothelial-blood cell signaling. Finally, to test if PM2 performs the therapeutic function of Activated Protein C (APC) that has been reported to stimulate its cytosolic effects via the β-arrestin pathway, we knocked down β-arrestin in the endothelium and analyzed clot formation again. We found that clotting reoccurred in vascular channel, thus showing that PM2 acts like an APC-like drug candidate. In conclusion, the Lung Alveolus-on-a-chip reconstitutes organ-level responses to blood clotting and may offer a valuable platform for drug development by allowing to dissect contributions of various cells in their mechanism of action.
