Metabolic markers detect early ostedifferentiation of mesenchymal stem cells from multiple donors

resumo

Background Mesenchymal stem cells (MSC) are pivotal bioengineering tools, offering significant promise for applications in bone regeneration. However, their therapeutic potential is limited by inter-donor variability and experimental issues. This study aimed to identify robust metabolic markers of osteodifferentiation applicable across multiple donors, while providing insight into the metabolic pathways actively involved in the process. Methods Untargeted nuclear magnetic resonance (NMR) metabolomics was applied to characterize the intra- and extracellular metabolic adaptations of human adipose-derived MSC (hAMSC) undergoing osteogenic differentiation, compared to proliferation alone. Multivariate and univariate statistical analysis was carried out on data from three independent donors, and cross-validation was employed to evaluate the predictive capacity of the proposed markers. Results Variations in the levels of selected (nine) intracellular and (seventeen) extracellular metabolites detect osteodifferentiation by day 7 (out of 21), with nearly 100% accuracy. These signatures suggest a metabolic shift from glycolysis/OxPhos to lactic fermentation, fatty acid β-oxidation and phosphocreatine hydrolysis. Intracellular glucose, lactate, citrate and specific amino acids are redirected towards protein synthesis and glycosylation, with some of the secreted metabolites (e.g., citrate) seemingly involved in biomineralization and other extracellular roles. Membrane metabolism, antioxidant mechanisms and adenosine metabolism are also impacted by osteodifferentiation. Conclusions These findings reveal effective donor-independent markers of hAMSC osteodifferentiation, with a robust extracellular signature standing out for potential rapid and non-invasive detection of osteocommitted cells.

autores

Daniela S. C. Bispo; Inês C. R. Graça; Catarina S. H. Jesus; João E. Rodrigues; Marlene C. Correia; Sabrina Atella; Iola F. Duarte; Brian J. Goodfellow; Mariana B. Oliveira; João F. Mano; Ana M. Gil

Grupos

G5 - Materiais Biomiméticos, Biológicos e Vivos

Projectos

CICECO - Aveiro Institute of Materials (UIDB/50011/2020)

CICECO - Aveiro Institute of Materials (UIDP/50011/2020)

Associated Laboratory CICECO-Aveiro Institute of Materials (LA/P/0006/2020)

Metabolite-activated 3D stem cell differentiation into bone (BetterBone)

Advancing the Regenerative and Translational Potential of Cellular Fibers (CellFi)

agradecimentos

This work was developed within the scope of the CICECO-Aveiro Institute of Materials, UIDB/50011/2020 project (doi: https://doi.org/10.54499/UIDB/50011/2020), UIDP/50011/2020 (doi: https://doi.org/10.54499/UIDP/50011/2020) and LA/P/0006/2020 (doi:https://doi.org/10.54499/LA/P/0006/2020), financed by national funds through the FCT/MCTES (PIDDAC). We acknowledge funds from the Foundation for Science and Technology through the BetterBone project (2022.04286.PTDC, doi: https://doi.org/10.54499/2022.04286.PTDC), the Portuguese National NMR Network (RNRMN), supported by Infrastructure Project Nº 022161 (co-financed by FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC); and FCT/SPQ PhD grant (DSCB) (SFRH/BD/150655/2020, doi: https://doi.org/10.54499/SFRH/BD/150655/2020). This work was also supported by the Programa Operacional Competitividade e Internacionalização, in the component FEDER, and by national funds (OE) through FCT/MCTES, in the scope of the project “CellFi”, PTDC/BTM-ORG/3215/2020 (DOI10.54499/PTDC/BTM-ORG/3215/2020).