Abstract
Bone fracture risk increases with age, disease states, and with use of certain therapeutics, such as acid-suppressive drugs, steroids and high-dose bisphosphonates. Historically, investigations into factors that underlie bone fracture risk have focused on evaluation of bone mineral density (BMD). However, numerous studies have pointed to factors other than BMD that contribute to fragility, including changes in bone collagen and water. The goal of this study is to investigate the feasibility of using near infrared spectral imaging (NIRSI) to determine the spatial distribution and relative amount of water and organic components in whole cross-sections of bone, and to compare those results to those obtained using magnetic resonance imaging (MRI) methods. Cadaver human whole-section tibiae samples harvested from 18 donors of ages 27-97years underwent NIRSI and ultrashort echo time (UTE) MRI. As NIRSI data is comprised of broad absorbances, second derivative processing was evaluated as a means to narrow peaks and obtain compositional information. The (inverted) second derivative peak heights of the NIRSI absorbances correlated significantly with the mean peak integration of the water, collagen and fat NIR absorbances, respectively, indicating that either processing method could be used for compositional assessment. The 5797cm(-1) absorbance was validated as arising from the fat present in bone marrow, as it completely disappeared after ultrasonication. The MRI UTE-determined bound water content in tibial cortical bone samples ranged from 62 to 91%. The NIRSI water peaks at 5152cm(-1) and at 7008cm(-1) correlated significantly with the UTE data, with r=0.735, p=0.016, and r=0.71, p=0.0096, respectively. There was also a strong correlation between the intensity of the NIRSI water peak at 7008cm(-1) and the intensity of the collagen peak at 4608cm(-1) (r=0.69, p=0.004). Since NIRSI requires minimal to no sample preparation, this approach has great potential to become a gold standard modality for the investigation of changes in water content, distribution, and environment in pre-clinical studies of bone pathology and therapeutics.