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Osteoporosis is a bone disease characterized by low bone mass and deterioration of the tissue leading to increased fragility. Osteopontin (OPN), a noncollageneous bone matrix protein, has been shown to play an important role in osteoporosis, bone resorption, and mineralization. However, OPN's role in bone mechanical properties on the submicron scale has not been studied in any detail. In this study, nanoindentation techniques were utilized to investigate how OPN and aging affect bone mechanical properties. Hardness and elastic modulus were calculated and compared between the OPN-deficient mice (OPN(-/-)) and their age and sex-matched wild-type (OPN(+/+)) controls. The results show that the mechanical properties of the young OPN(-/-) bones (age < 12 weeks) are significantly lower than that of the youngest OPN(+/+) bones. This finding was confirmed by additional microindentation testing. Biochemical analysis using micro-Raman spectroscopy indicated more mineral content in young OPN(+/+) bones. Older (age > 12 weeks) bones did not show any significant differences in mechanical properties with genotype. In addition, OPN(+/+) bones show a decrease in mechanical properties between young and older age groups. By contrast, OPN(-/-) bones showed no significant change in mechanical properties with aging.

Pseudoxanthoma elasticum (PXE) is a heritable multisystem disorder characterized by ectopic mineralization. However, the structure of the mineral deposits, their interactions with the connective tissue matrix, and the details of the progressive maturation of the mineral crystals are currently unknown. In this study, we examined the mineralization processes in Abcc6−/− mice, a model system for PXE, by energy dispersive X-ray, and Fourier transform infrared imaging spectroscopy (FT-IRIS). The results indicated that the principal components of the mineral deposits were calcium and phosphate which co-localized within the histologically demonstrable lesions determined by topographic mapping. The Ca/P ratio increased in samples with progressive mineralization reaching the value comparable to that in endochondral bone. A progressive increase in mineralization was also reflected by increased mineral-to-matrix ratio determined by FT-IRIS. Determination of the mineral phases by FT-IRIS suggested progressive maturation of the mineral deposits from amorphous calcium phosphate to hydroxyapatite. These results provide critical information of the mechanisms of mineralization in PXE, with potential pharmacologic implications.

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