Characterization of Microstructural Changes on Biglycan Induced Mice Bone by Low-Field Nuclear Magnetic Resonance

A NMR spin-spin (T₂) relaxation technique has been described for determining the porosity, and the bound water distribution in biglycan induced mouse bone and correlate to their mechanical properties. The technique of low-field proton NMR involves spin-spin relaxation and free induction decay (FID) measurements, and the computational inversion methods for decay data analysis. The CPMG T2 relaxation data can be inverted to T₂ relaxation distribution and this distribution then can be transformed to a pore size distribution with the longer relaxation times corresponding to larger pores. The FID T₂ relaxation data of dried bone (mobile water removed) can be inverted to T₂ relaxation distribution and this distribution then can be transformed to bound and solid-like water distribution with the longest relaxation time corresponding to bound water component. These techniques are applied to quantify apparent changes in porosity, and bound water in controlled and biglycan knockout mouse bone. Overall bone porosity from CPMG T₂ relaxation is determined using the calibrated NMR fluid volume from the proton relaxation data divided by overall bone volume. Ignore the physical sample differences, from the inversion FID T₂ relaxation spectrum, the ratio of the bound to solid-like water components is used to calibrate the bound water inside bone, and the results can be used to correlated bone mechanical properties. Hydration status significantly affects the toughness of bone, and bound water has been considered as a biomarker for prediction of bone fragility fractures. In addition to the collagen phase, recent evidence shows that glycosaminoglycans (GAGs) of proteoglycans (PGs) in the extracellular matrix also play a pivotal role in regulating the tissue-level hydration status of bone, there by affecting the tissue-level toughness of bone. Furthermore, biglycan and decorin are two major types of PGs in bone reports. Biglycan knockout induced changes in GAGs, bound water, as well as bone tissue toughness. Among all subtypes of PGs, biglycan is identified as a major subtype in the bone mineral matrix. In this study, we used a biglycan mouse model and the obtained bone samples were measured by low-field NMR to determine the bone porosity and bound water changes, and used to predict if knockout of biglycan may affect the amount of bound water and subsequently lead to reduce toughness of bone.