Cellulose-based scaffolds for fluorescence lifetime imaging-assisted tissue engineering

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dc.contributor.author O'Donnell, Neil
dc.contributor.author Okkelman, Irina A.
dc.contributor.author Timashev, Peter
dc.contributor.author Gromovykh, Tatyana I.
dc.contributor.author Papkovsky, Dmitri B.
dc.contributor.author Dmitriev, Ruslan I.
dc.date.accessioned 2018-11-09T11:20:34Z
dc.date.available 2018-11-09T11:20:34Z
dc.date.issued 2018-09-25
dc.identifier.citation O'Donnell, N., Okkelman, I. A., Timashev, P., Gromovykh, T. I., Papkovsky, D. B. and Dmitriev, R. I. (2018) 'Cellulose-based scaffolds for fluorescence lifetime imaging-assisted tissue engineering', Acta Biomaterialia, 80, pp. 85-96. doi:10.1016/j.actbio.2018.09.034 en
dc.identifier.volume 80 en
dc.identifier.startpage 85 en
dc.identifier.endpage 96 en
dc.identifier.issn 1742-7061
dc.identifier.issn 1878-7568
dc.identifier.uri http://hdl.handle.net/10468/7080
dc.identifier.doi 10.1016/j.actbio.2018.09.034
dc.description.abstract Quantitative measurement of pH and metabolite gradients by microscopy is one of the challenges in the production of scaffold-grown organoids and multicellular aggregates. Herein, we used the cellulose-binding domain (CBD) of the Cellulomonas fimi CenA protein for designing biosensor scaffolds that allow measurement of pH and Ca2+ gradients by fluorescence intensity and lifetime imaging (FLIM) detection modes. By fusing CBD with pH-sensitive enhanced cyan fluorescent protein (CBD-ECFP), we achieved efficient labeling of cellulose-based scaffolds based on nanofibrillar, bacterial cellulose, and decellularized plant materials. CBD-ECFP bound to the cellulose matrices demonstrated pH sensitivity comparable to untagged ECFP (1.9–2.3 ns for pH 6–8), thus making it compatible with FLIM-based analysis of extracellular pH. By using 3D culture of human colon cancer cells (HCT116) and adult stem cell-derived mouse intestinal organoids, we evaluated the utility of the produced biosensor scaffold. CBD-ECFP was sensitive to increases in extracellular acidification: the results showed a decline in 0.2–0.4 pH units in response to membrane depolarization by the protonophore FCCP. With the intestinal organoid model, we demonstrated multiparametric imaging by combining extracellular acidification (FLIM) with phosphorescent probe-based monitoring of cell oxygenation. The described labeling strategy allows for the design of extracellular pH-sensitive scaffolds for multiparametric FLIM assays and their use in engineered live cancer and stem cell-derived tissues. Collectively, this research can help in achieving the controlled biofabrication of 3D tissue models with known metabolic characteristics. Statement of Significance: We designed biosensors consisting of a cellulose-binding domain (CBD) and pH- and Ca2+-sensitive fluorescent proteins. CBD-tagged biosensors efficiently label various types of cellulose matrices including nanofibrillar cellulose and decellularized plant materials. Hybrid biosensing cellulose scaffolds designed in this study were successfully tested by multiparameter FLIM microscopy in 3D cultures of cancer cells and mouse intestinal organoids. en
dc.description.sponsorship Russian Science Foundation (Grant 18-15-00407); Ministry of Education and Science of the Russian Federation (Russian Academic Excellence Project 5-100) en
dc.format.mimetype application/pdf en
dc.language.iso en en
dc.publisher Elsevier Ltd. en
dc.rights © 2018, Elsevier Ltd. All rights reserved. This manuscript version is made available under the CC-BY-NC-ND 4.0 license. en
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/ en
dc.subject Biomaterials en
dc.subject Biosensor en
dc.subject FLIM en
dc.subject Live cell imaging en
dc.subject Organoid en
dc.subject Scaffold en
dc.title Cellulose-based scaffolds for fluorescence lifetime imaging-assisted tissue engineering en
dc.type Article (peer-reviewed) en
dc.internal.authorcontactother Ruslan Dmitriev, Biochemistry , University College Cork, Cork, Ireland. +353-21-490-3000 Email: r.dmitriev@ucc.ie en
dc.internal.availability Full text available en
dc.check.info Access to this article is restricted until 24 months after publication by request of the publisher. en
dc.check.date 2020-09-25
dc.date.updated 2018-11-09T10:18:41Z
dc.description.version Accepted Version en
dc.internal.rssid 461216052
dc.contributor.funder Science Foundation Ireland en
dc.contributor.funder Russian Science Foundation en
dc.contributor.funder Ministry of Education and Science of the Russian Federation en
dc.description.status Peer reviewed en
dc.identifier.journaltitle Acta Biomaterialia en
dc.internal.copyrightchecked Yes en
dc.internal.licenseacceptance Yes en
dc.internal.IRISemailaddress r.dmitriev@ucc.ie en
dc.relation.project info:eu-repo/grantAgreement/SFI/SFI Starting Investigator Research Grant (SIRG)/13/SIRG/2144/IE/Development of Bionic Sensor Materials for Metabolic Imaging in Regenerative Medicine/ en
dc.relation.project info:eu-repo/grantAgreement/SFI/SFI Research Centres/12/RC/2276/IE/I-PIC Irish Photonic Integration Research Centre/ en

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© 2018, Elsevier Ltd. All rights reserved. This manuscript version is made available under the CC-BY-NC-ND 4.0 license. Except where otherwise noted, this item's license is described as © 2018, Elsevier Ltd. All rights reserved. This manuscript version is made available under the CC-BY-NC-ND 4.0 license.
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