Direct monitoring of single-cell response to biomaterials by Raman spectroscopy

Abstract There is continued focus on the development of new biomaterials and associated biological testing methods needed to reduce the time taken for their entry to clinical use. The application of Raman spectroscopy to the study of individual cells that have been in contact with biomaterials offer...

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Autores principales:	Mary Josephine McIvor, Preetam K. Sharma, Catherine E. Birt, Hayley McDowell, Shannon Wilson, Stephen McKillop, Jonathan G. Acheson, Adrian R. Boyd, Brian J. Meenan
Formato:	article
Lenguaje:	EN
Publicado:	Springer 2021
Materias:	Materials of engineering and construction. Mechanics of materials TA401-492 Medical technology R855-855.5
Acceso en línea:	https://doaj.org/article/e48ddc9a41cf4c53a39cc90d6d948e3f
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spelling	oai:doaj.org-article:e48ddc9a41cf4c53a39cc90d6d948e3f2021-12-05T12:19:16ZDirect monitoring of single-cell response to biomaterials by Raman spectroscopy10.1007/s10856-021-06624-50957-45301573-4838https://doaj.org/article/e48ddc9a41cf4c53a39cc90d6d948e3f2021-12-01T00:00:00Zhttps://doi.org/10.1007/s10856-021-06624-5https://doaj.org/toc/0957-4530https://doaj.org/toc/1573-4838Abstract There is continued focus on the development of new biomaterials and associated biological testing methods needed to reduce the time taken for their entry to clinical use. The application of Raman spectroscopy to the study of individual cells that have been in contact with biomaterials offers enhanced in vitro information in a potentially non-destructive testing regime. The work presented here reports the Raman spectral analysis of discreet U-2 OS bone cells after exposure to hydroxyapatite (HA) coated titanium (Ti) substrates in both the as-deposited and thermally annealed states. These data show that cells that were in contact with the bioactive HA surface for 7 days had spectral markers similar to those cultured on the Ti substrate control for the same period. However, the spectral features for those cells that were in contact with the annealed HA surface had indicators of significant differentiation at day 21 while cells on the as-deposited surface did not show these Raman changes until day 28. The cells adhered to pristine Ti control surface showed no spectral changes at any of the timepoints studied. The validity of these spectroscopic results has been confirmed using data from standard in vitro cell viability, adhesion, and proliferation assays over the same 28-day culture period. In this case, cell maturation was evidenced by the formation of natural bone apatite, which precipitated intracellularly for cells exposed to both types of HA-coated Ti at 21 and 28 days, respectively. The properties of the intracellular apatite were markedly different from that of the synthetic HA used to coat the Ti substrate with an average particle size of 230 nm, a crystalline-like shape and Ca/P ratio of 1.63 ± 0.5 as determined by SEM-EDX analysis. By comparison, the synthetic HA particles used as a control had an average size of 372 nm and were more-rounded in shape with a Ca/P ratio of 0.8 by XPS analysis and 1.28 by SEM-EDX analysis. This study shows that Raman spectroscopy can be employed to monitor single U-2 OS cell response to biomaterials that promote cell maturation towards de novo bone thereby offering a label-free in vitro testing method that allows for non-destructive analyses.Mary Josephine McIvorPreetam K. SharmaCatherine E. BirtHayley McDowellShannon WilsonStephen McKillopJonathan G. AchesonAdrian R. BoydBrian J. MeenanSpringerarticleMaterials of engineering and construction. Mechanics of materialsTA401-492Medical technologyR855-855.5ENJournal of Materials Science: Materials in Medicine, Vol 32, Iss 12, Pp 1-18 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Materials of engineering and construction. Mechanics of materials TA401-492 Medical technology R855-855.5
spellingShingle	Materials of engineering and construction. Mechanics of materials TA401-492 Medical technology R855-855.5 Mary Josephine McIvor Preetam K. Sharma Catherine E. Birt Hayley McDowell Shannon Wilson Stephen McKillop Jonathan G. Acheson Adrian R. Boyd Brian J. Meenan Direct monitoring of single-cell response to biomaterials by Raman spectroscopy
description	Abstract There is continued focus on the development of new biomaterials and associated biological testing methods needed to reduce the time taken for their entry to clinical use. The application of Raman spectroscopy to the study of individual cells that have been in contact with biomaterials offers enhanced in vitro information in a potentially non-destructive testing regime. The work presented here reports the Raman spectral analysis of discreet U-2 OS bone cells after exposure to hydroxyapatite (HA) coated titanium (Ti) substrates in both the as-deposited and thermally annealed states. These data show that cells that were in contact with the bioactive HA surface for 7 days had spectral markers similar to those cultured on the Ti substrate control for the same period. However, the spectral features for those cells that were in contact with the annealed HA surface had indicators of significant differentiation at day 21 while cells on the as-deposited surface did not show these Raman changes until day 28. The cells adhered to pristine Ti control surface showed no spectral changes at any of the timepoints studied. The validity of these spectroscopic results has been confirmed using data from standard in vitro cell viability, adhesion, and proliferation assays over the same 28-day culture period. In this case, cell maturation was evidenced by the formation of natural bone apatite, which precipitated intracellularly for cells exposed to both types of HA-coated Ti at 21 and 28 days, respectively. The properties of the intracellular apatite were markedly different from that of the synthetic HA used to coat the Ti substrate with an average particle size of 230 nm, a crystalline-like shape and Ca/P ratio of 1.63 ± 0.5 as determined by SEM-EDX analysis. By comparison, the synthetic HA particles used as a control had an average size of 372 nm and were more-rounded in shape with a Ca/P ratio of 0.8 by XPS analysis and 1.28 by SEM-EDX analysis. This study shows that Raman spectroscopy can be employed to monitor single U-2 OS cell response to biomaterials that promote cell maturation towards de novo bone thereby offering a label-free in vitro testing method that allows for non-destructive analyses.
format	article
author	Mary Josephine McIvor Preetam K. Sharma Catherine E. Birt Hayley McDowell Shannon Wilson Stephen McKillop Jonathan G. Acheson Adrian R. Boyd Brian J. Meenan
author_facet	Mary Josephine McIvor Preetam K. Sharma Catherine E. Birt Hayley McDowell Shannon Wilson Stephen McKillop Jonathan G. Acheson Adrian R. Boyd Brian J. Meenan
author_sort	Mary Josephine McIvor
title	Direct monitoring of single-cell response to biomaterials by Raman spectroscopy
title_short	Direct monitoring of single-cell response to biomaterials by Raman spectroscopy
title_full	Direct monitoring of single-cell response to biomaterials by Raman spectroscopy
title_fullStr	Direct monitoring of single-cell response to biomaterials by Raman spectroscopy
title_full_unstemmed	Direct monitoring of single-cell response to biomaterials by Raman spectroscopy
title_sort	direct monitoring of single-cell response to biomaterials by raman spectroscopy
publisher	Springer
publishDate	2021
url	https://doaj.org/article/e48ddc9a41cf4c53a39cc90d6d948e3f
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Direct monitoring of single-cell response to biomaterials by Raman spectroscopy

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