Dual NMR for probing intrinsic bone structure and a potential gut‐bone axis in ovariectomized rats

Currently the existence of a gut-bone axis receives massive attention, and while sound premises and indirect proofs exist for the gut-bone axis concept, few studies have provided actual data linking the gut and bone physically. This study aimed to exploit the versatile nature of nuclear magnetic resonance (NMR) to link NMR relaxometry data on bone mineralization with NMR spectroscopic profiling of gut metabolites. For this purpose, sample material was obtained from a 6-week intervention study with ovariectomized (OVX) rats (n=49) fed with 7 different diets varying in calcium content (0.2-6.0 mg/Kg) and prebiotic fiber content (0-5.0 % w/w). This design ensured a span in i) calcium available for bone mineralization and ii) metabolic activity in the gut. After termination of the intervention, longitudinal (T1), transverse (T2) relaxation as well as mechanical bone strength were measured on the excised femur bones. A PLS model with high predictability (Q2=0.86, R2=0.997) was demonstrated between T2 decay curves and femur mechanical strength. Correlations were established between bone T2 populations and gut short-chain fatty acids. In conclusion, the present dual NMR approach showed strong correlation between T2 relaxation and mechanical strength of the bone, and when metabolic activity in the gut was modulated by inulin, the potential existence of a gut-bone axis was demonstrated.

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