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• W2337752224 abstract "Event Abstract Back to Event Intrinsically degradable protein patterns for temporal stem cell engineering Bethany Almeida1 and Anita Shukla1 1 Brown University, School of Engineering, Center for Biomedical Engineering, United States Introduction: Stem cell geometry has been linked to differentiation[1],[2]. We hypothesize that the effects of geometric confinement on stem cells is time dependent. To study this hypothesis, we developed intrinsically degradable protein patterns. Briefly, alkanethiol self-assembled monolayer (SAM) patterns were deposited on gold (Au) of various thicknesses. Fibronectin was then adsorbed to these patterns (Fig 1A). Degradation of these surfaces was tested in various physiological environments (e.g. cell culture media) and characterized using physical and chemical methods. Differentiation of human mesenchymal stem cells (HMSCs) on these surfaces was investigated. Methods: Fabrication and Characterization: Hexadecanethiol (HT) and 2-{2-[2-(2-{2-[2-(1-mercaptoundec-11-yloxy)-ethoxy]-ethoxy}-ethoxy)-ethoxy]-ethoxy}-ethanol thiols (OEG6) (4 mM) were applied to Au-coated glass. Characterization methods included atomic force microscopy (AFM), matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometry, and x-ray photoelectron spectroscopy. Cell Culture and Differentiation: HMSCs were cultured in 1:1 v/v osteogenic:adipogenic differentiation media. Lipids were stained with oil red O, and calcification was measured by an alkaline phosphatase activity assay. Results and Discussion: We examined the physical changes of our SAM surfaces in cell culture media to understand behavior in cell culture conditions. An increase in roughness 24 hours after HT deposition indicates protein adsorption. At day 5, 4 nm Au showed increased roughness (Fig 1B), suggesting Au mottling[3]. HT-Au, 8 nm (Fig 1D) and 10 nm (Fig 1F), show a decrease in roughness over time, suggesting desorption of HT and proteins likely due to metal oxidation. Au thickness affected stability; 8 and 10 nm Au showed greater stability than 4 nm Au. We examined the chemical changes associated with degradation to confirm that the mechanism of degradation was Au oxidation and mottling. MALDI-TOF peaks for thiol, glycol, and Au fragments were seen in 4, 8, and 10 nm Au-OEG6 on days 5, 7, and 5, respectively, further indicating Au mottling and OEG6 desorption. XPS results demonstrated a decrease of carbon-sulfur and increase in carbon over time (Fig 1C,E,G), while the total Au did not change, indicating that degradation was driven primarily by oxidation. Finally, cells confined by these substrates demonstrated increased levels of lipid formation as compared to non-confined controls (Fig 2), which continued to increase over time as the patterned surfaces underwent degradation for all Au thicknesses examined; there was no change in calcification. Conclusions: The degradation behavior of these Au-SAM surfaces may be used in stem cell engineering. Our studies confirmed that Au oxidation and mottling cause pattern degradation, dependent on Au thickness. Confinement on our substrates affected stem cell differentiation in a time dependent manner. Shukla LaboratoryReferences:[1] Kilian, K. a, Bugarija, B., Lahn, B. T., & Mrksich, M. (2010). Geometric cues for directing the differentiation of mesenchymal stem cells. Proceedings of the National Academy of Sciences of the United States of America, 107(11), 4872–7. http://doi.org/10.1073/pnas.0903269107[2] Mrksich, M., Dike, L. E., Tien, J., Ingber, D. E., & Whitesides, G. M. (1997). Using microcontact printing to pattern the attachment of mammalian cells to self-assembled monolayers of alkanethiolates on transparent films of gold and silver. Experimental Cell Research, 235(2), 305–13. http://doi.org/10.1006/excr.1997.3668[3] Ostuni, E., et al., Using self-assembled monolayers to pattern ECM proteins and cells on substrates. Methods Mol Biol, 2009. 522: p. 183-94. Keywords: Cell Differentiation, stem cell, Surface modification, matrix-cell interaction Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Biomaterials for cellular programming Citation: Almeida B and Shukla A (2016). Intrinsically degradable protein patterns for temporal stem cell engineering. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.00874 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 Bethany Almeida Anita Shukla Google Bethany Almeida Anita Shukla Google Scholar Bethany Almeida Anita Shukla PubMed Bethany Almeida Anita Shukla 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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• W2337752224 title "Intrinsically degradable protein patterns for temporal stem cell engineering" @default.
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