NMR Metabolomic Analysis of Osteoblasts Pre and Post Differentiation

Additional contributors: Louis S. Kidder; Bruce E. Hammer (faculty mentor).The research is relevant to 18 million Americans, primarily the elderly, who suffer from osteopenia (loss of bone mass) and osteoporosis. The other targeted population is astronauts, in low-earth orbit, who lose 1 to 3 % of their bone mass per month. 1H-1DNMR (700MHz) spectra of water soluble extracts from MC3T3 Osteoblastic cells were obtained. Two osteoblast populations were studied – one cultured in cell media with the addition of ascorbic acid and the other population cultured in cell media without the addition of ascorbic. Ascorbic acid induces osteoblast cells to differentiate resulting in the production of bone. The metabolomic signature of osteoblast cells, pre and post differentiation, were compared through 1H-1DNMR spectroscopy . 1H NMR Spectra was analyzed by Chenomx 6.1 software which is available at the University of Minnesota Biomedical NMR facility. Chenomx deconvolves the chemical shifts of an NMR spectrum and assigns them to known metabolite signatures. This allows the identification of roughly 100 to 200 metabolites. Chemical shifts that Chenomx did not have in its library was assigned using 1H-2D-NOSEY NMR spectroscopy. The techniques developed here will be used in future experiments for cells that are magnetically levitated to observe changes in cell metabolomics when gravitational loading is reduced or eliminated.This project continuation is supported by the Undergraduate Research Opportunities Program (UROP) and computational resources from Minnesota Supercomputing Institute for Advanced Computational Research of the University of Minnesota