NMR Metabolomics Assessment of Osteogenic Differentiation of Adipose-Tissue-Derived Mesenchymal Stem Cells

abstract

This Article presents, for the first time to our knowledge, an untargeted nuclear magnetic resonance (NMR) metabolomic characterization of the polar intracellular metabolic adaptations of human adipose-derived mesenchymal stem cells during osteogenic differentiation. The use of mesenchymal stem cells (MSCs) for bone regeneration is a promising alternative to conventional bone grafts, and untargeted metabolomics may unveil novel metabolic information on the osteogenic differentiation of MSCs, allowing their behavior to be understood and monitored/guided toward effective therapies. Our results unveiled statistically relevant changes in the levels of just over 30 identified metabolites, illustrating a highly dynamic process with significant variations throughout the whole 21-day period of osteogenic differentiation, mainly involving amino acid metabolism and protein synthesis; energy metabolism and the roles of glycolysis, the tricarboxylic acid cycle, and oxidative phosphorylation; cell membrane metabolism; nucleotide metabolism (including the specific involvement of O-glycosylation intermediates and NAD(+)); and metabolic players in protective antioxidative mechanisms (such as glutathione and specific amino acids). Different metabolic stages are proposed and are supported by putative biochemical explanations for the metabolite changes observed. This work lays the groundwork for the use of untargeted NMR metabolomics to find potential metabolic markers of osteogenic differentiation efficacy.

keywords

BONE-MARROW; OSTEOBLAST; MINERALIZATION; PROLIFERATION; ADIPOGENESIS; SPECTROSCOPY; OSTEOPONTIN; METABOLISM; MECHANISMS; RESORPTION

subject category

Biochemistry & Molecular Biology

authors

Bispo, DSC; Jesus, CSH; Correia, M; Ferreira, F; Bonifazio, G; Goodfellow, BJ; Oliveira, MB; Mano, JF; Gil, AM

Groups

G5 - Biomimetic, Biological and Living Materials

Projects

A Metabolomics-guided Bioreactor for Improved Engineered Bone Implants (BioImplant)

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

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

Rede Nacional de Ressonância Magnética Nuclear (PTNMR)

acknowledgements

We acknowledge the Portuguese Foundation for Science and Technology (FCT) for cofunding the BIOIMPLANT project (PTDC/BTM-ORG/28835/2017) through the COMPETE2020 program and European Union fund FEDER (POCI-01-0145-FEDER-028835). C.S.H.J. is grateful to the same project for funding their contracts with the University of Aveiro. D.S.C.B. acknowledges the Sociedade Portuguesa de Quimica and FCT for her Ph.D. grant SFRH/BD/150655/2020. All authors are grateful to the CICECO-Aveiro Institute of Materials project, with references UIDB/50011/2020 and UIDP/50011/2020, financed by national funds through the FCT/MEC and when appropriate cofinanced by FEDER under the PT2020 Partnership Agreement. The NMR spectrometer used in this work is part of the National NMR Network (PTNMR) and is partially supported by infrastructure project no. 022161 (cofinanced by FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC).