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Online quantitative monitoring of live cell engineered cartilage growth using diffuse fiber-optic Raman spectroscopy

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Title: Online quantitative monitoring of live cell engineered cartilage growth using diffuse fiber-optic Raman spectroscopy
Authors: Bergholt, M
Albro, M
Stevens, MM
Item Type: Journal Article
Abstract: Tissue engineering (TE) has the potential to improve the outcome for patients with osteoarthritis (OA). The successful clinical translation of this technique as part of a therapy requires the ability to measure extracellular matrix (ECM) production of engineered tissues in vitro, in order to ensure quality control and improve the likelihood of tissue survival upon implantation. Conventional techniques for assessing the ECM content of engineered cartilage, such as biochemical assays and histological staining are inherently destructive. Raman spectroscopy, on the other hand, represents a non-invasive technique for in situ biochemical characterization. Here, we outline current roadblocks in translational Raman spectroscopy in TE and introduce a comprehensive workflow designed to non-destructively monitor and quantify ECM biomolecules in large (>3 mm), live cell TE constructs online. Diffuse near-infrared fiber-optic Raman spectra were measured from live cell cartilaginous TE constructs over a 56-day culturing period. We developed a multivariate curve resolution model that enabled quantitative biochemical analysis of the TE constructs. Raman spectroscopy was able to non-invasively quantify the ECM components and showed an excellent correlation with biochemical assays for measurement of collagen (R2 = 0.84) and glycosaminoglycans (GAGs) (R2 = 0.86). We further demonstrated the robustness of this technique for online prospective analysis of live cell TE constructs. The fiber-optic Raman spectroscopy strategy developed in this work offers the ability to non-destructively monitor construct growth online and can be adapted to a broad range of TE applications in regenerative medicine toward controlled clinical translation.
Issue Date: 14-Jun-2017
Date of Acceptance: 12-Jun-2017
URI: http://hdl.handle.net/10044/1/49140
DOI: https://dx.doi.org/10.1016/j.biomaterials.2017.06.015
ISSN: 1878-5905
Publisher: Elsevier
Start Page: 128
End Page: 137
Journal / Book Title: Biomaterials
Volume: 140
Copyright Statement: © 2017 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Sponsor/Funder: Medical Research Council (MRC)
Commission of the European Communities
Medical Research Council (MRC)
Commission of the European Communities
Funder's Grant Number: MR/K026682/1
ERC-2013-CoG-616417
RA26F7
701713
Keywords: Articular cartilage
Fiber-optic Raman spectroscopy
Live cell Raman spectroscopy
Online biomedical Raman spectroscopy
Tissue-engineering
Biomedical Engineering
MD Multidisciplinary
Publication Status: Published
Open Access location: http://www.sciencedirect.com/science/article/pii/S0142961217304088
Appears in Collections:Faculty of Engineering
Materials
Faculty of Natural Sciences



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