ORCID
- Francesca Ravera: 0000-0002-8393-2668
- Esen Efeoglu: 0000-0002-3380-0499
- Hugh J Byrne: 0000-0002-1735-8610
Abstract
Raman microspectroscopy (RMS) offers a powerful, non-destructive approach for in situ monitoring of dynamic biochemical processes within cells. However, the ability to reliably data-mine the spectroscopic signatures and their evolution and extract meaningful information can be challenging. Multivariate Curve Resolution-Alternating Least Squares (MCR-ALS) regression analysis is a powerful chemometric technique that can potentially address this challenge by deconvoluting the spectra into individual component spectra, each representing a specific biochemical species, and regressing the solutions against kinetic constraints. In this study, MCR-ALS analysis was performed on spectral data of differentiation process of mesenchymal stem cells into chondrocytes, carried out on two different substrates, collagen 3-dimensional hydrogel and the conventional 2-dimensional culture model at time intervals of 1-7, 14, 21 days. The kinetic evolution of the chondrogenesis was modelled according to a phenomenological rate equation model, in an attempt to describe the biochemical evolution of the cell composition within the process. Moreover, the ability of the algorithm to faithfully extract the correct reaction rates and spectral profiles has been explored. The results indicated that the differentiation process originates in the nucleolar regions, subsequently extending to the nuclear and cytoplasmic compartments and corroborated a more rapid differentiation rate in cell cultures grown on 3D collagen hydrogels compared to 2D substrates. The combination of Raman microspectroscopy and MCR-ALS offers a powerful approach for elucidating the complex mechanisms underlying chondrogenesis and developing innovative strategies for regenerative medicine.
DOI Link
Publication Date
2025-07-21
Publication Title
Analyst
Volume
150
Issue
15
ISSN
0003-2654
First Page
3445
Last Page
3456
Acceptance Date
2025-01-01
Deposit Date
2026-08-28
Funding
FR acknowledges financial support from the TU Dublin Fiosraigh Postgraduate Scholarship programme.
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
Additional Links
Recommended Citation
Ravera, Francesca; Efeoglu, Esen; and Byrne, Hugh J, "Monitoring the kinetic evolution of mesenchymal stem cell differentiation using Raman microspectroscopy" (2025). Research Outputs: 2025-Present. 8.
https://arrow.tudublin.ie/focas2ro/8