Cortical bone material / compositional properties in growing children and young adults aged 1.5-23 years, as a function of gender, age, metabolic activity, and growth spurt

Abstract

Bone material / compositional properties are significant determinants of bone quality, thus strength. Raman spectroscopic analysis provides information on the quantity and quality of all three bone tissue components (mineral, organic matrix, and tissue water). The overwhelming majority of the published reports on the subject concern adults. We have previously reported on these properties in growing children and young adults, in the cancellous compartment. The purpose of the present study was to create normative reference data of bone material / compositional properties for children and young adults, in the cortical compartment. We performed Raman (Senterra (Bruker Optik GmbH), 50× objective, with an excitation of 785 nm (100 mW) and a lateral resolution of ~0.6 μm) microspectroscopic analysis of transiliac bone samples from 54 individuals between 1.5 and 23 years of age, with no known metabolic bone disease, and which have been previously used to establish histomorphometric, bone mineralization density distribution, and cancellous bone quality reference values. The bone quality indices that were determined were: mineral/matrix ratio (MM) from the integrated areas of the v₂PO₄ (410-460 cm^-1) and the amide III (1215-1300 cm^-1) bands, tissue water in nanopores approximated by the ratio of the integrated spectral area ~ 494-509 cm^-1 to Amide III band, the glycosaminoglycan (GAG) content (ratio of integrated area 1365-1390 cm^-1 to the Amide III band, the sulfated proteoglycan (sPG) content as the ratio of the integrated peaks ~1062 cm^-1 and 1365-1390 cm^-1, the pyridinoline (Pyd) content estimated from the ratio of the absorbance height at 1660 cm^-1 / area of the amide I (1620-1700 cm^-1) band, and the mineral maturity / crystallinity (MMC) estimated from the inverse of the full width at half height of the v₁PO₄ (930-980 cm^-1) band. Analyses were performed at the three distinct cortical surfaces (endosteal, osteonal, periosteal) at specific anatomical microlocations, namely the osteoid, and the three precisely known tissue ages based on the presence of fluorescence double labels. Measurements were also taken in interstitial bone, a much older tissue that has undergone extensive secondary mineralization. Overall, significant dependencies of the measured parameters on tissue age were observed, while at any given tissue age, sex and subject age were minimal confounders. The established Raman database in the cortical compartments complements the previously published one in cancellous bone, and provides healthy baseline bone quality indices that may serve as a valuable tool to identify alterations due to pediatric disease.

Keywords: Bone material properties; Cortical bone; Healthy children; Raman spectroscopy; Tissue age.