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• W2326291698 abstract "Event Abstract Back to Event Quantifying the mechanics of idiopathic pulmonary fibrosis in 3D microculture systems Avital Horowitz1, David Habiel2, Cory Hogaboam3, Shuichi Takayama4 and Christopher Moraes1 1 McGill University, Chemical Engineering, Canada 2 Cedars-Sinai Medical Center, Medicine, United States 3 University of Michigan Medical School, Pathology, United States 4 University of Michigan College of Engineering, Biomedical Engineering, United States Introduction: Idiopathic pulmonary fibrosis (IPF) is a chronic and fatal disease characterized by progressive stiffening of the lung matrix[1]. IPF is perpetuated by pathologically activated fibroblasts that are significantly more contractile than normal lung fibroblasts (NLFs)[2], however the underlying mechanisms of IPF remain poorly understood. Our objective in this work is to quantify the dynamics of healthy and diseased primary fibroblast cell-matrix interactions in physiologically-realistic, but precisely-defined culture systems. Understanding the biomechanics underlying IPF disease progression will provide insight to potential drug targets and therapies. Materials and Methods: A collagen microgel contraction assay was previously developed to rapidly measure cell contractile forces[3] (Fig 1A). The micro-scale platform circumvents transport limitations inherent in conventional bulk culture systems, enabling direct comparisons between cell types. An aqueous two-phase system consisting of cells/collagen/dextran phase and a polyethylene glycol phase is used to print a microgel in a cell culture environment. Remodelling of the microgels by normal and IPF patient-derived primary fibroblasts cells is monitored optically to measure cellular contraction. Collagen contraction experiments were followed with single-cell traction force microscopy (TFM) using standard techniques[4]. Results: The collagen contraction assay indicated that NLF and IPF cells were indistinguishable in terms of contractile remodelling in the presence of fetal bovine serum (FBS). In contrast, serum-free culture showed marked increases in contractile activity for IPF patient samples (Fig1B). Since NLFs are FBS-sensitive and IPF contraction levels remain consistently high, this suggests that high contractility is a pathological baseline state in IPF. Furthermore, since serum abrogates these differences, and IPF cells are inherently more contractile than healthy cells[1],[5], this suggests that serum-supplemented cultures may not provide an adequate platform to study IPF. Although collagen contraction assays are fast and simple, collagen structure and geometry can be unstable providing only a rough estimate of cell forces[6]. Thus we confirmed the results obtained by measuring forces at the cellular level using TFM. Preliminary analysis suggests that NLFs exert greater traction forces in the presence of serum compared to serum-starvation conditions (Fig2), and that IPF cell traction forces were not affected by serum (data not shown). Conclusion: The results suggest that IPF fibroblast dynamics are different, and distinctly regulated from healthy lung fibroblasts. Moreover, there appears to be no functional difference between healthy and IPF dynamics in regularly supplemented serum. Future experimentation will explore the effects of various soluble factors on healthy and IPF dynamics in order to further model microscale IPF physiology. This work supported by a Natural Sciences and Engineering Research Council of Canada Discovery Grant and Undergraduate Student Research Award, and a Province of Quebec Fonds Nature et Technologies New Investigator award.References:[1] Marinković, A., Mih, J. D., Park, J. A., Liu, F. & Tschumperlin, D. J. Improved throughput traction microscopy reveals pivotal role for matrix stiffness in fibroblast contractility and TGF-β responsiveness. American Journal of Physiology - Lung Cellular and Molecular Physiology 303, L169-180, doi:10.1152/ajplung.00108.2012 (2012).[2] Marinkovic, A., Liu, F. & Tschumperlin, D. J. Matrices of physiologic stiffness potently inactivate idiopathic pulmonary fibrosis fibroblasts. American journal of respiratory cell and molecular biology 48, 422-430, doi:10.1165/rcmb.2012-0335OC (2013).[3] Moraes, C., Simon, A. B., Putnam, A. J. & Takayama, S. Aqueous two-phase printing of cell-containing contractile collagen microgels. Biomaterials 34, 9623-9631, doi:10.1016/j.biomaterials.2013.08.046 (2013).[4] Style, R. W. et al. Traction force microscopy in physics and biology. Soft matter 10, 4047-4055, doi:10.1039/c4sm00264d (2014).[5] Parker, M. W. et al. Fibrotic extracellular matrix activates a profibrotic positive feedback loop. The Journal of clinical investigation 124, 1622-1635, doi:10.1172/jci71386 (2014).[6] Li, B. & Wang, J. H. C. Fibroblasts and myofibroblasts in wound healing: Force generation and measurement. Journal of Tissue Viability 20, 108-120, doi:http://dx.doi.org/10.1016/j.jtv.2009.11.004 (2011). Keywords: Hydrogel, in vitro, cell, biomaterial Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Microdevices: reproducing physiology at microscale Citation: Horowitz A, Habiel D, Hogaboam C, Takayama S and Moraes C (2016). Quantifying the mechanics of idiopathic pulmonary fibrosis in 3D microculture systems. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.02902 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 27 Mar 2016; Published Online: 30 Mar 2016. Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Avital Horowitz David Habiel Cory Hogaboam Shuichi Takayama Christopher Moraes Google Avital Horowitz David Habiel Cory Hogaboam Shuichi Takayama Christopher Moraes Google Scholar Avital Horowitz David Habiel Cory Hogaboam Shuichi Takayama Christopher Moraes PubMed Avital Horowitz David Habiel Cory Hogaboam Shuichi Takayama Christopher Moraes Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page." @default.
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• W2326291698 title "Quantifying the mechanics of idiopathic pulmonary fibrosis in 3D microculture systems" @default.
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