abstract
Mesenchymal stem cell (MSC) osteodifferentiation is accompanied by important lipid metabolic adaptations, which may reveal relevant biomarkers and potential osteoinductive species. However, high donor variability remains a challenge for biomarker identification. This work unveiled shared lipid features of human adipose-tissue MSC (hAMSC) for three independent donors, using an untargeted NMR spectroscopy methodology. The results showed that osteodifferentiation induced increases in esterified cholesterol preferentially enriched in shorter monounsaturated fatty acids (MUFA), and triacylglycerides containing longer fatty acids (FA), both consistent with increased lipid droplet formation in the cytosol. Membrane adaptations involved hydrolysis of phosphatidylcholine (PtdCho) and phosphatidylethanolamine (PtdEtn), possibly to allow subsequent polyunsaturated FA incorporation (to enhance membrane fluidity) and facilitate removal of peroxidized FA, while originating inorganic phosphate (Pi) for mineralization. PtdCho levels seem closely linked to the creatine-phosphocreatine axis, reflecting a shared contribution to Pi generation. MUFA also appeared to serve as preferential substrates for β-oxidation, apparently in association with antioxidative mechanisms. The above metabolic effects were indicative of a common pathway modulation in the three donors, with predicted upregulation of ALP, collagen, antioxidant enzymes, LDL, and HSP27, and downregulation of ERK 1/2 (notably upregulated at day 7), arginase, and vacuolar H+-ATPase. Notably, the proposed donor-independent lipid signature enabled the detection of osteodifferentiating cells with nearly 90 % accuracy, highlighting the complex interplay among different lipid families, with esterified cholesterol, triacylglycerides, and phospholipids emerging as main players in osteodifferentiation.
authors
Daniela S.C. Bispo; Inês C.R. Graça; Catarina S.H. Jesus; João E. Rodrigues; Brian J. Goodfellow; Mariana B. Oliveira; João F. Mano; Ana M. Gil
our authors
Ana Maria Pissarra Coelho Gil
Associate Professor with Aggregation
Brian James Goodfellow
Assistant Professor
Catarina Sofia Henriques de Jesus
Junior Researcher
Daniela Bispo
PhD Student
Inês C. R. Graça
Research Fellowship
João A. Rodrigues
Junior Researcher
João Mano
Full professor
Mariana Braga de Oliveira
Assistant ResearcherProjects
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)
acknowledgements
This work was developed within the scope of the CICECO-Aveiro Institute of Materials, UIDB/50011/2020 project (doi: 10.54499/UIDB/50011/2020), UIDP/50011/2020 (doi:10.54499/UIDP/50011/2020) and LA/P/0006/2020 (doi: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: 10.54499/2022.04286.PTDC), the Portuguese National NMR Network (RNRMN), supported by Infrastructure Project N° 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: 10.54499/SFRH/BD/150655/2020).