Label-Free Evaluation of Osteogenic Differentiation Using Extended-Focus Raman Imaging

Xi, Heqi; Bhongsatiern, Phan; Li, Menglu; Iwayama, Tomoaki; Takedachi, Masahide; Yamaguchi, Yoshinori; Fujita, Katsumasa

doi:https://doi.org/10.1021/acs.analchem.5c00921

Label-Free Evaluation of Osteogenic Differentiation Using Extended-Focus Raman Imaging

Xi, Heqi; Bhongsatiern, Phan; Li, Menglu et al.

Analytical Chemistry, 2025, 97(39), 21218-21228

Number of Access:6（2025-11-25 23:45 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/103020

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Title	Label-Free Evaluation of Osteogenic Differentiation Using Extended-Focus Raman Imaging
Creator	CreatorName Xi, Heqi
	CreatorName Bhongsatiern, Phan
	CreatorName Li, Menglu
	NRID 1000080757865 1000080757865 ORCID 0000-0002-3405-5822 0000-0002-3405-5822 researchmap permalink iwayamat iwayamat Scopus Author ID 36916715900 36916715900 CreatorName Iwayama, Tomoaki
	NRID 1000060452447 1000060452447 researchmap permalink takedachi_m takedachi_m Scopus Author ID 6507251637 6507251637 CreatorName Takedachi, Masahide
	NRID 1000020386634 1000020386634 CreatorName Yamaguchi, Yoshinori
	NRID 1000080362664 1000080362664 ORCID 0000-0002-2284-375X 0000-0002-2284-375X researchmap permalink read0089911 read0089911 Scopus Author ID 7404058829 7404058829 CreatorName Fujita, Katsumasa
Description	This document is the Accepted Manuscript version of a Published Work that appeared in final form in Analytical Chemistry, © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acs.analchem.5c00921
Abstract	Periodontal ligament stem cell (PDLSC)-based therapy has emerged as a promising approach to regenerate damaged periodontal tissue in periodontitis treatments. To advance the application of this therapy, a nondestructive method to monitor osteogenic differentiation and evaluate the osteogenic abilities of PDLSCs is essential. In this study, we developed a rapid extended-focus Raman imaging technique for the label-free evaluation of cell differentiation. The developed technique enabled the high-throughput acquisition of Raman spectra of the whole cell volume, facilitating the monitoring of osteogenic differentiation of PDLSCs lineages with low and high osteogenic abilities. Using multivariate analysis of Raman spectra, we identified extracellular matrix (ECM) components represented by collagen type I as key contributors to the spectral differences between two cell populations. Combining Raman imaging and fluorescence observation, we confirmed the correlation between ECM deposition and hydroxyapatite (HA) generation, which indicated that ECM accumulation reflects the osteogenic ability of PDLSCs. These findings highlight the application of the extended-focus Raman imaging approach to realize the label-free assessment of PDLSC lineages with distinct osteogenic capabilities, paving the way to nondestructive cell screening in periodontal tissue regeneration treatment.
Publisher	American Chemical Society
Source Title	Analytical Chemistry
Volume (Issue)	97(39)
Page	21218-21228
Date of Issued	2025-09-24
Language	English
Handle URL	https://hdl.handle.net/11094/103020
PISSN	00032700
eISSN	15206882
Relation.isVersionOf	DOI (Publisher Version) https://doi.org/10.1021/acs.analchem.5c00921
Access Rights	embargoed access
CopyRight	The full-text file will be made open to the public on 24 September 2026 in accordance with the publisher's policy.
oaire:version	Accepted Manuscript
Category	学術雑誌論文 Journal Article

Resource Type	journal article
Local Type	Journal Article
DCMI Type	Text
DCTERMS.bibliographicCitation	Analytical Chemistry. 2025, 97(39), p. 21218-21228
DC.title	Label-Free Evaluation of Osteogenic Differentiation Using Extended-Focus Raman Imaging
DC.creator	Xi, Heqi
	Bhongsatiern, Phan
	Li, Menglu
	Iwayama, Tomoaki
	Takedachi, Masahide
	Yamaguchi, Yoshinori
	Fujita, Katsumasa
DC.publisher	American Chemical Society
DC.language' scheme='DCTERMS.RFC1766	English
DC.type' scheme='DCTERMS.DCMIType	Text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2025-09-24
DC.identifier	https://doi.org/10.1021/acs.analchem.5c00921
DC.description	This document is the Accepted Manuscript version of a Published Work that appeared in final form in Analytical Chemistry, © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acs.analchem.5c00921
DCTERMS.abstract	Periodontal ligament stem cell (PDLSC)-based therapy has emerged as a promising approach to regenerate damaged periodontal tissue in periodontitis treatments. To advance the application of this therapy, a nondestructive method to monitor osteogenic differentiation and evaluate the osteogenic abilities of PDLSCs is essential. In this study, we developed a rapid extended-focus Raman imaging technique for the label-free evaluation of cell differentiation. The developed technique enabled the high-throughput acquisition of Raman spectra of the whole cell volume, facilitating the monitoring of osteogenic differentiation of PDLSCs lineages with low and high osteogenic abilities. Using multivariate analysis of Raman spectra, we identified extracellular matrix (ECM) components represented by collagen type I as key contributors to the spectral differences between two cell populations. Combining Raman imaging and fluorescence observation, we confirmed the correlation between ECM deposition and hydroxyapatite (HA) generation, which indicated that ECM accumulation reflects the osteogenic ability of PDLSCs. These findings highlight the application of the extended-focus Raman imaging approach to realize the label-free assessment of PDLSC lineages with distinct osteogenic capabilities, paving the way to nondestructive cell screening in periodontal tissue regeneration treatment.
DC.rights	The full-text file will be made open to the public on 24 September 2026 in accordance with the publisher's policy.
DC.format	103020
citation_title	Label-Free Evaluation of Osteogenic Differentiation Using Extended-Focus Raman Imaging
citation_author	Xi, Heqi
	Bhongsatiern, Phan
	Li, Menglu
	Iwayama, Tomoaki
	Takedachi, Masahide
	Yamaguchi, Yoshinori
	Fujita, Katsumasa
citation_publisher	American Chemical Society
citation_language	English
citation_date	2025-09-24
citation_journal_title	Analytical Chemistry
citation_volume	97
citation_issue	39
citation_firstpage	21218
citation_lastpage	21228
citation_doi	https://doi.org/10.1021/acs.analchem.5c00921
citation_public_url	https://hdl.handle.net/11094/103020

Label-Free Evaluation of Osteogenic Differentiation Using Extended-Focus Raman Imaging

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