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

dc.check.date2020-09-25
dc.check.infoAccess to this article is restricted until 24 months after publication by request of the publisher.en
dc.contributor.authorO'Donnell, Neil
dc.contributor.authorOkkelman, Irina A.
dc.contributor.authorTimashev, Peter
dc.contributor.authorGromovykh, Tatyana I.
dc.contributor.authorPapkovsky, Dmitri B.
dc.contributor.authorDmitriev, Ruslan I.
dc.contributor.funderScience Foundation Irelanden
dc.contributor.funderRussian Science Foundationen
dc.contributor.funderMinistry of Education and Science of the Russian Federationen
dc.date.accessioned2018-11-09T11:20:34Z
dc.date.available2018-11-09T11:20:34Z
dc.date.issued2018-09-25
dc.date.updated2018-11-09T10:18:41Z
dc.description.abstractQuantitative 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.sponsorshipRussian Science Foundation (Grant 18-15-00407); Ministry of Education and Science of the Russian Federation (Russian Academic Excellence Project 5-100)en
dc.description.statusPeer revieweden
dc.description.versionAccepted Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationO'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.2018.09.034en
dc.identifier.doi10.1016/j.actbio.2018.09.034
dc.identifier.endpage96en
dc.identifier.issn1742-7061
dc.identifier.issn1878-7568
dc.identifier.journaltitleActa Biomaterialiaen
dc.identifier.startpage85en
dc.identifier.urihttps://hdl.handle.net/10468/7080
dc.identifier.volume80en
dc.language.isoenen
dc.publisherElsevier Ltd.en
dc.relation.projectinfo: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.projectinfo:eu-repo/grantAgreement/SFI/SFI Research Centres/12/RC/2276/IE/I-PIC Irish Photonic Integration Research Centre/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.urihttps://creativecommons.org/licenses/by-nc-nd/4.0/en
dc.subjectBiomaterialsen
dc.subjectBiosensoren
dc.subjectFLIMen
dc.subjectLive cell imagingen
dc.subjectOrganoiden
dc.subjectScaffolden
dc.titleCellulose-based scaffolds for fluorescence lifetime imaging-assisted tissue engineeringen
dc.typeArticle (peer-reviewed)en
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