期刊
BIOMATERIALS
卷 140, 期 -, 页码 128-137出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.biomaterials.2017.06.015
关键词
Fiber-optic Raman spectroscopy; Articular cartilage; Tissue-engineering; Live cell Raman spectroscopy; Online biomedical Raman spectroscopy
资金
- UK Regenerative Medicine Platform Hub Acellular Approaches for Therapeutic Delivery [MR/K026682/1]
- UK Regenerative Medicine Platform Hub A Hub for Engineering and Exploiting the Stem Cell Niche [MR/K026666/1]
- grant State of the Art Biomaterials Development and Characterization of the Cell-Biomaterial Interface from MRC [MR/ L012677/1]
- Medical Engineering Solutions in the Osteoarthritis Centre of Excellence
- Wellcome Trust
- Engineering and Physical Sciences Research Council [088844]
- ERC [616417]
- H2020 through Individual Marie Sklodowska-Curie Fellowship IMAGINE [701713]
- MRC [MR/K026666/1, MR/L012677/1] Funding Source: UKRI
- European Research Council (ERC) [616417] Funding Source: European Research Council (ERC)
- Marie Curie Actions (MSCA) [701713] Funding Source: Marie Curie Actions (MSCA)
- Medical Research Council [MR/K026666/1, MR/L012677/1] Funding Source: researchfish
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 (R-2 = 0.84) and glycosaminoglycans (GAGS) (R-2 = 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. (C) 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/).
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