¹H-NMR-based metabolomics reveals amino acid metabolic preferences and a shared metabolic axis across osteoblastic cell lines

Commercially available osteoblastic cell lines have been widely applied in bone research, yet their metabolic characteristics and metabolic remodeling during osteogenic differentiation remain poorly defined. In this study, 1 H-NMR-based metabolomic technique was used to detect and compare metabolic characteristics and osteogenic phenotypes of three classic osteoblastic cell lines, including MG63, hFOB, and MC3T3. Phenotypic results revealed significant differences in proliferation capacity and osteogenic potential among the three cell lines following osteogenic induction. Metabolomic KEGG enrichment analysis revealed that MG63 was mainly enriched in branched-chain amino acid metabolism, hFOB was primarily enriched in glycine, serine, and threonine metabolism, while MC3T3 showed dominant enrichment in arginine and proline metabolism. Exogenous supplementation of the corresponding amino acids (branched-chain amino acids for MG63; glycine for hFOB; arginine for MC3T3) enhanced osteogenic capacity in the respective cell lines. Further cross-group metabolomic analysis revealed that the alanine, aspartate, and glutamate metabolic pathway was commonly enriched across the three cell lines. Notably, exogenous glutamate supplementation negatively regulated osteogenic differentiation in these cell lines. Overall, this study began with metabolomic screening and discovered the relationships between amino acid-related metabolic remodeling and osteogenic differentiation across the three osteoblastic cell lines. These findings may provide candidate metabolic signatures for further investigations in basic bone biology research.