abstract
Strategies involving mesenchymal stem cell (MSC) osteogenic differentiation are currently promising tools in regenerative medicine. However, there is a need to further understand the biochemical processes underlying differentiation, in order to optimize their osteogenic commitment. Several intra- and extracellular events, typically occurring over a period of 21 days of osteogenic differentiation, are known to determine osteogenic efficiency, namely protein synthesis and transport to the extracellular matrix (ECM), and mineralization in the ECM, in tandem with several intracellular metabolic adaptations [1,2]. The use of vibrational spectroscopic techniques with high sensitivity and specificity is known to unveil exquisite detail on subcellular events triggered by different stimuli, e.g. carcinogenesis or drug treatment [3,4]. However, such techniques have been underexplored to follow osteogenic differentiation [5]. This work presents a systematic study by complementary vibrational spectroscopy (FTIR and Raman) of human adipose tissue MSC (hAMSC) across multiple timepoints of osteodifferentiation, with particular focus on biomolecular changes taking place at the intra- and extracellular levels. Along a 21-day period, hAMSC under osteogenic differentiation were compared to the corresponding control samples (absence of osteogenetic stimuli, only cell proliferation occurring). Results revealed at day 21 the appearance of hydroxyapatite features, being a marker of osteogenesis [5]. Additionally, cellular adaptation during osteogenesis was monitored through changes in membrane fluidity and cytoskeleton/ECM structural characteristics.Overall, our results demonstrate the promise of subcellular insights of osteogenesis as provided by vibrational spectroscopy in tandem with microscopy. This knowledge will support the definition of informed strategies towards optimization of MSC osteodifferentiation, in biomedical applications.
authors
Mariela M. Nolasco, João A. Rodrigues, Clara B. Martins, Ana L.M. Batista de Carvalho, Daniela S. C. Bispo, Helena I.S. Nogueira, Paulo J.A. Ribeiro-Claro, Mariana B. Oliveira, João F. Mano, Maria P.M. Marques, Ana M. Gil
our authors
Ana Maria Pissarra Coelho Gil
Associate Professor with Aggregation
Daniela Bispo
PhD Student
Helena Isabel Seguro Nogueira
Assistant Professor with Aggregation
João A. Rodrigues
Junior Researcher
João Mano
Full professor
Mariana Braga de Oliveira
Assistant Researcher
Mariela Martins Nolasco
Assistant Researcher
Paulo Ribeiro-Claro
Associate Professor with AggregationGroups
G1 - Porous Materials and Nanosystems
G5 - Biomimetic, Biological and Living Materials
G6 - Virtual Materials and Artificial Intelligence
Projects
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
BetterBone (2022.04286.PTDC, doi: 10.54499/2022.04286.PTDC); CICECO-Aveiro Institute of Materials UIDB/50011/2020 (doi: 10.54499/ UIDB/50011/2020), UIDP/50011/2020 (doi: 10.54499/UIDP/50011/2020) & LA/P/0006/2020 (doi: 10.54499/LA/P/0006/2020); SFRH/BD/150655/2020; QFM-UC (Coimbra) UIDB/00070/2020 (https://doi.org/10.54499/UIDB/00070/2020) and UIDP/00070/2020 (https://doi.org/10.54499/ UIDP/00070/2020). Portuguese National NMR Network (Project Nº022161) through FEDER (COMPETE 2020/POCI/PORL/FCT (PIDDAC).